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系統識別號 U0007-0207200902272700
論文名稱(中文) 比較兩非麥角多巴胺受體致效劑的小型第四期臨床試驗: 帕金森氏症病人由力必平劑量逐步調整為樂伯克之評估
論文名稱(英文) A Small-scale Phase IV Clinical Trial of Two Non-ergot Dopamine Agonists: Slowly Switching from Ropinirole to Pramipexole in Patients with Parkinson's Disease
校院名稱 臺北醫學大學
系所名稱(中) 藥學研究所
系所名稱(英) Graduate Institute of Pharmacy
學年度 97
學期 2
出版年 98
研究生(中文) 蔡欣佳
學號 M301096022
學位類別 碩士
語文別 英文
口試日期 2009-06-15
論文頁數 65頁
口試委員 委員-陳榮基
委員-許秀蘊
委員-闕壯卿
共同指導教授-袁瑞昱
指導教授-劉景平
關鍵字(中) 多巴胺第三亞型受體
帕金森氏症
第四期臨床試驗
樂伯克
力必平
帕金森氏症狀衡量表(UPDRS)
關鍵字(英) D3 receptor
Parkinson's disease
Phase IV clinical trial
Pramipexole
Ropinirole
UPDRS
學科別分類
中文摘要 研究背景:
多巴胺第二亞型受體致效劑 (DAs)可作為早期帕金森氏症單獨治療方式,或因長期使用左多巴(L-DOPA)而產生併發症的晚期病人之輔助治療。 DAs依化學結構分為麥角及非麥角。 最近研究指出,使用高劑量麥角類DAs會增加心臟瓣膜疾病發生率。 因此,由麥角換成非麥角DAs以預防此類副作用為近期研究目標。 然而,非麥角DAs間的直接替換較少被研究,因此力必平(Requip®)及樂伯克(Mirapex®)的轉換因子多由間接比較所得,數值也不盡相同。 樂伯克與力必平不同之處在於樂伯克有較高的生體可用率及較長的半衰期。 樂伯克不經肝臟代謝酵素系統代謝也不與此系統有相互作用,腎臟為其主要排除方式。 此外,樂伯克對於多巴胺第三亞型受體有較高的親和力,以及具有可能潛在的神經保護效果。 這些性質使得樂伯克具有額外的抗憂鬱作用。

研究目的:
本研究主旨是探討台灣帕金森氏症病人由力必平換成樂伯克後所得的轉換因子,並評估樂伯克可能具有的抗憂鬱效果。

研究方法:
本研究為前瞻性開放標籤的小型第四期臨床試驗,在臺北醫學大學附設醫院進行為期一年之研究(九十七年六月到九十八年六月)。 帕金森病人(四十五歲到八十歲)參與此試驗前已服用固定劑量的力必平或併服左多巴達四週者,可加入此研究。 病人停用力必平後,以劑量逐步調整方式換成樂伯克。 當達到與換藥前相同效果之劑量時,即以此穩定有效劑量維持十二週。 病人換藥前為自己換藥後的對照組,此前後測的配對實驗可減少干擾因子,因此小樣本數即可用於測試結果。 主要結果依據帕金森氏症狀衡量表(UPDRS-III)評估動作功能,探討十二週後力必平與樂伯克的轉換因子。 次要結果包括換藥前後貝氏憂鬱量表(BDI-II)及帕金森生活品質量表(PDQ-39)的分數改變。 此外,病人主觀滿意度(TSQM)及換藥期間安全性也納入評估。

研究結果:
共十七位病人參與此研究,其中十位完成此試驗。 換藥前力必平平均每天劑量為3.1毫克,換藥後樂伯克平均每天劑量為0.6 毫克。 依每位病人結果所得轉換因子為5.6±2.8。 帕金森氏症狀衡量表從22.4±10.2分降為15.8±6.6分,顯示動作功能進步。 貝氏憂鬱量表分數在換藥期間無顯著改變,而帕金森生活品質量表及藥物治療滿意程度量表的效果及整體滿意度兩部分,在維持穩定有效劑量第二個月時均有明顯分數改變。 暈眩及便秘為換藥期間常見副作用。

研究結論:
在維持樂伯克穩定有效劑量十二星期後,所得轉換因子為5.6時動作有明顯進步。 此轉換因子可應用於台灣帕金森氏症患者由力必平隔夜迅速換成樂伯克時所需劑量的計算。
英文摘要 Background:
Dopamine receptor agonists (DAs), especially D2 subtype could be used as monotherapy in patients with early Parkinson’s disease (PD) or adjunctive therapy to L-DOPA in advanced PD with motor complications. DAs are divided into ergot and non-ergot agents. Recent studies indicate that taking high dosages of ergot-derived DAs are associated with valvular heart disease. Therefore, switching from ergot to non-ergot DAs to prevent such adverse drug reactions is the main research goal of many clinical trials. To the best of our knowledge, very few studies have investigated the direct switch between non-ergot DAs. The conversion factor between ropinirole and pramipexole is calculated indirectly and thus the published values are inconsistent. Pramipexole is a non-ergot DA, which has more bioavailability (> 90% vs. 50%) and longer half-life (8~12 hours vs. 6 hours) than ropinirole. Moreover, pramipexole is not metabolized by cytochrome P450 enzymes and it does not have interactions with this enzyme system; pramipexole is primarily eliminated through the kidneys. Furthermore, pramipexole has been shown to have relatively high affinity for the D3 receptor and putative neuroprotective effects. These properties of pramipexole may lead to additional antidepressant action.

Objectives:
The mission of this study was to determine the conversion factor between two non-ergot DAs, ropinirole (Requip®) and pramipexole (Mirapex®) in Taiwanese PD patients and evaluated the possible antidepressant effect of pramipexole.

Methods:
In this prospective open-label pilot study, a small-scale phase IV clinical trial of two non-ergot DAs was carried out at Taipei Medical University Hospital, during the period of June, 2008 to June, 2009. Patients with PD (aged 45 to 80 years old) had taken stable doses of ropinirole with or without L-DOPA (Madopar®) for at least 28 days were recruited. After stopping ropinirole treatment, patients were switched to pramipexole with a log escalating dose until reaching the optimal motor control. The effective dosage of pramipexole was kept unchanged in the three-month maintenance period. This paired design would minimize confounding factors and thus a small sample size was needed for studying the primary and secondary endpoints. The primary endpoint was to determine the conversion factor between ropinirole and pramipexole at the end of the 12-week maintenance period, assessed by the Unified Parkinson’s Disease Rating Scale, part III (UPDRS-III). The secondary outcomes included changes in the Beck Depression Inventory-Second Edition (BDI-II) and the 39-item Parkinson's Disease Questionnaire (PDQ-39) scores during the study period. Patient’s satisfaction with pramipexole by Treatment Satisfaction Questionnaire for Medication (TSQM) and safety of the switch were also evaluated.

Results:
A total of 17 PD patients were recruited and 10 of them completed the 12-week maintenance course. The pre-switch dosage of ropinirole was 3.1 mg/day and the post-switch maintenance dosage of pramipexole was 0.6 mg/day. The dosage of pramipexole was lower than that in the western countries. The conversion factor was 5.6±2.8 (n=10), that was directly derived from each patient’s paired data. The UPDRS-III score showed a significant decrease from 22.4±10.2 to 15.8±6.6 (p<0.05), reflecting an improvement in motor performance. There was no significant change in the BDI-II during the switch period. However, a significant difference in the PDQ-39 summary index was obtained after the second month of the maintenance period. There were significant changes in both effectiveness and global satisfaction scores of the TSQM simultaneously. The most common adverse events while switching to pramipexole were dizziness and constipation.

Conclusions:
In summary, a directly slow switch from ropinirole to pramipexole at a conversion factor of 5.6 showed the motor improvement after a 12-week maintenance period. This factor derived from the present study could be useful in developing a protocol of an overnight switch from ropinirole to pramipexole in Taiwanese patients with PD.
論文目次 List of Contents...I
Abstract...II
摘要...V
List of Tables...VII
List of Figures... VIII
List of Appendices...IX
Abbreviations...X
Text...XI

Table 1. DAs in the world market for idiopathic PD...10
Table 2. Pharmacodynamic and pharmacokinetic properties of the DAs...11
Table 3. Antiparkinsonian medications at Taipei Medical University Hospital...13
Table 4. Conversion chart for rapid switch among DAs and L-DOPA...19
Table 5. Summarized conversion factors between ropinirole (R) and pramipexole (P)...20
Table 6. Pre-switch characteristics of the recruited patients with PD...28
Table 7. Paired analysis of outcome measures on the primary and secondary endpoints...31
Table 8. Individual conversion factor after switching from ropinirole (R) to pramipexole (P)...33
Table 9A. Changes in outcome measures between the first and second months...36
Table 9B. Changes in outcome measures between the second and third months...37
Table 9C. Changes in outcome measures between the first and third months...38
Table 10. Patients’ satisfaction with pramipexole treatment ...39
Table 11. Adverse events while switching to pramipexole..41

Figure 1. Patients with PD for a small-scale phase IV clinical trial: enrollment and dropouts...27
Figure 2. Changes in the UPDRS-III after switching from ropinirole to pramipexole in patients with PD...32
Figure 3. Changes in the BDI-II during the switch period.34
Figure 4. Changes in the PDQ-39 summary index while patients were switched from ropinirole to pramipexole...35

Appendix A. The Institutional Review Board Approval
Letter...64

Chapter I.Preface...1
1.1 Rationale...1
1.2 Working Hypothesis...2
1.3 Research Goals...2
Chapter II. Literature Review...4
2.1 Epidemiology...4
2.2 Clinical Presentation and Diagnosis...5
2.2.1 Etiology and Pathophysiology...5
2.2.2 Diagnosis... 5
2.2.3 Clinical Stages...5
2.3 Treatment Guidelines and Recommendations...6
2.4 Overview of Pharmacologic Therapy...6
2.4.1 Levodopa...6
2.4.2 Dopamine Receptor Agonists...8
2.4.3 Other Pharmacologic Agents...12
2.5 Clinical Issues...14
2.5.1 Cardiac Problems Caused by Ergot-derived DAs...14
2.5.2 Non-motor Symptom of PD: Depression...14
2.5.3 Resting Tremor...15
2.6. Clinical Trials of Switching Dopamine Agonists...16
2.7 Briefing of New Discovery...21
Chapter III. Methods...22
3.1 Patients...22
3.2 Study Design and Outcomes...23
3.3 Statistical Analysis... 25
Chapter IV. Results...26
4.1 Patients...26
4.2 Efficacy...29
4.3 Adverse Events...40
Chapter V. Discussion...42
5.1 Primary Endpoint...42
5.2 Secondary Endpoints...46
5.3 Adverse Events ...47
5.4 Cost...48
5.5 Limitations... 48
Chapter VI. Conclusions...50
References...51


參考文獻 Atkinson MJ, Sinha A, Hass SL, Colman SS, Kumar RN, Brod M and Rowland CR (2004) Validation of a general measure of treatment satisfaction, the Treatment Satisfaction Questionnaire for Medication (TSQM), using a national panel study of chronic disease. Health Qual Life Outcomes 2:12.
Barone P, Scarzella L, Marconi R, Antonini A, Morgante L, Bracco F, Zappia M and Musch B (2006) Pramipexole versus sertraline in the treatment of depression in Parkinson's disease: a national multicenter parallel-group randomized study. J Neurol 253:601-607.
Beck AT and Steer RA (1984) Internal consistencies of the original and revised Beck Depression Inventory. J Clin Psychol 40:1365-1367.
Bernheimer H, Birkmayer W, Hornykiewicz O, Jellinger K and Seitelberger F (1973) Brain dopamine and the syndromes of Parkinson and Huntington. Clinical, morphological and neurochemical correlations. J Neurol Sci 20:415-455.
Brown RG, MacCarthy B, Gotham AM, Der GJ and Marsden CD (1988) Depression and disability in Parkinson's disease: A follow-up of 132 cases. Psychol Med 18:49-55.
Calne DB (1993) Treatment of Parkinson's disease. N Engl J Med 329:1021-1027.
Carlsson A, Lindqvist M and Magnusson T (1957) 3,4-Dihydroxyphenylalanine and 5-hydroxytryptophan as reserpine antagonists. Nature 180:1200.
Chen JJ (2007) Optimizing use of a dopamine agonist in Parkinson’s disease. Pharmacy Times:111-119.
Chen JJ and Swope DM (2007) Pharmacotherapy for Parkinson's disease. Pharmacotherapy 27:161S-173S.
Chen RC, Chang SF, Su CL, Chen THH, Yen MF, Wu HM, Chen ZY and Liou HH (2001) Prevalence, incidence, and mortality of PD: A door-to-door survey in Ilan County, Taiwan. Neurology 57:1679-1686.
Cockcroft DW and Gault MH (1976) Prediction of creatinine clearance from serum creatinine. Nephron 16:31-41.
Corrigan MH, Denahan AQ, Wright CE, Ragual RJ and Evans DL (2000) Comparison of pramipexole, fluoxetine, and placebo in patients with major depression. Depress Anxiety 11:58-65.
Costa A, Peppe A, Dell'Agnello G, Caltagirone C and Carlesimo GA (2009) Dopamine and cognitive functioning in de novo subjects with Parkinson's disease: Effects of pramipexole and pergolide on working memory. Neuropsychologia 47:1374-1381.
Cotzias GC, Papavasiliou PS and Gellene R (1969) Modification of Parkinsonism--chronic treatment with L-dopa. N Engl J Med 280:337-345.
Cotzias GC, Van Woert MH and Schiffer LM (1967) Aromatic amino acids and modification of parkinsonism. N Engl J Med 276:374-379.
Cummings JL (1992) Depression and Parkinson's disease: A review. Am J Psychiatry 149:443-454.
Deleu D, Northway MG and Hanssens Y (2002) Clinical pharmacokinetic and pharmacodynamic properties of drugs used in the treatment of Parkinson's disease. Clin Pharmacokinet 41:261-309.
Deuschl G, Schade-Brittinger C, Krack P, Volkmann J, Schafer H, Botzel K, Daniels C, Deutschlander A, Dillmann U, Eisner W, Gruber D, Hamel W, Herzog J, Hilker R, Klebe S, Kloss M, Koy J, Krause M, Kupsch A, Lorenz D, Lorenzl S, Mehdorn HM, Moringlane JR, Oertel W, Pinsker MO, Reichmann H, Reuss A, Schneider GH, Schnitzler A, Steude U, Sturm V, Timmermann L, Tronnier V, Trottenberg T, Wojtecki L, Wolf E, Poewe W and Voges J (2006) A randomized trial of deep-brain stimulation for Parkinson's disease. N Engl J Med 355:896-908.
Dorsey ER, Constantinescu R, Thompson JP, Biglan KM, Holloway RG, Kieburtz K, Marshall FJ, Ravina BM, Schifitto G, Siderowf A and Tanner CM (2007) Projected number of people with Parkinson disease in the most populous nations, 2005 through 2030. Neurology 68:384-386.
Etminan M, Gill S and Samii A (2003) Comparison of the risk of adverse events with pramipexole and ropinirole in patients with Parkinson's disease: a meta-analysis. Drug Saf 26:439-444.
Fahn S (2008) The history of dopamine and levodopa in the treatment of Parkinson's disease. Mov Disord 23 Suppl 3:S497-508.
Fahn S and Elton RL (1987) Unified Parkinson’s Disease Rating Scale In: Fahn S, Marsden CD, Calne D, Goldstein M, editors. Recent developments in Parkinson’s disease, Vol. 2. Florham Park, N.J: MacMillan Health Care Information.153-163.
Fahn S, Oakes D, Shoulson I, Kieburtz K, Rudolph A, Lang A, Olanow CW, Tanner C and Marek K (2004) Levodopa and the progression of Parkinson's disease. N Engl J Med 351:2498-2508.
Fernandez HH and Chen JJ (2007) Monoamine oxidase-B inhibition in the treatment of Parkinson's disease. Pharmacotherapy 27:174S-185S.
Gau C, Lii P and Hsu C (2008) Evidence to Support the Regulation Decision for the Usage of Pergolide in Patients with Parkinson’s Disease in Taiwan. Drug Saf 31:916-917.
Gibb WR and Lees AJ (1988) The relevance of the Lewy body to the pathogenesis of idiopathic Parkinson's disease. J Neurol Neurosurg Psychiatry 51:745-752.
Goetz CG, Blasucci L and Stebbins GT (1999) Switching dopamine agonists in advanced Parkinson's disease: Is rapid titration preferable to slow? Neurology 52:1227-1229.
Goetz CG, Poewe W, Rascol O and Sampaio C (2005) Evidence-based medical review update: Pharmacological and surgical treatments of Parkinson's disease: 2001 to 2004. Mov Disord 20:523-539.
Goetz CG, Shannon KM, Tanner CM, Carroll VS and Klawans HL (1989) Agonist substitution in advanced Parkinson's disease. Neurology 39:1121-1122.
Goetz CG, Tanner CM, Glantz RH and Klawans HL (1985) Chronic agonist therapy for Parkinson's disease: a 5-year study of bromocriptine and pergolide. Neurology 35:749-751.
Grosset K, Needleman F, Macphee G and Grosset D (2004) Switching from ergot to nonergot dopamine agonists in Parkinson's disease: A clinical series and five-drug dose conversion table. Mov Disord 19:1370-1374.
Grosset KA and Grosset DG (2006) Proposed dose equivalence for rapid switching between dopamine agonists in Parkinson's disease. Clin Ther 28:1063-1064; author reply 1064.
Hauser R, Beider C, Stacey M, Hubble J, Seeberger L and Gauger K (2000) Acute versus gradual pramipexole to ropinirole switch. Mov Disord 15:133.
Hauser RA, Rascol O, Korczyn AD, Stoessl AJ, Watts RL, Poewe W, De Deyn PP and Lang AE (2007) Ten-year follow-up of Parkinson's disease patients randomized to initial therapy with ropinirole or levodopa. Mov Disord 22:2409-2417.
Hobson DE, Lang AE, Wayne Martin WR, Razmy A, Rivest J and Fleming J (2002) Excessive daytime sleepiness and sudden-onset sleep in Parkinson disease: A survey by the Canadian Movement Disorders Group. JAMA 287:455-463.
Hoehn MM and Yahr MD (1967) Parkinsonism: onset, progression and mortality. Neurology 17:427-442.
Holloway R, Shoulson I, Kieburtz K, McDermott M, Tariot P, Kamp C, Day D, Shinaman A, Fahn S, Lang A, Marek K, Seibyl J, Weiner W, Welsh M, Pahwa R, Coe S, Barclay L, Sutherland L, Hildebrand K, Hubble J, Weeks C, Lewitt P, Miyasaki J, Duff J, Sime E, Suchowersky O, Stacy M, Kurth M, Brewer M, Harrigan M, Russell DS, Fussell B, Ford B, Dillon S, Hammerstad J, Stone C, Riley D, Rainey P, Standaert D, Tennis M, Wooten F, Rost-ruffner E, Factor S, Brown D, Jankovic J, Atassi F, Kurlan R, Gardiner I, Panisset M, Amyot D, Hall J, Rajput A, Shirley T, Rodnitzky R, Dobson J, Shults C, Fontaine D, Waters C, Schuman S, Pfeiffer R, Rast S, Pfeiffer B, Brocht A, Casaceli C, Daigneault S, Hodgeman K, Honsinger K, O'Connell C and Watts A (2000) Pramipexole vs Levodopa as initial treatment for Parkinson disease: A randomized controlled trial. JAMA 284:1931-1938.
Holloway RG, Shoulson I, Fahn S, Kieburtz K, Lang A, Marek K, McDermott M, Seibyl J, Weiner W, Musch B, Kamp C, Welsh M, Shinaman A, Pahwa R, Barclay L, Hubble J, LeWitt P, Miyasaki J, Suchowersky O, Stacy M, Russell DS, Ford B, Hammerstad J, Riley D, Standaert D, Wooten F, Factor S, Jankovic J, Atassi F, Kurlan R, Panisset M, Rajput A, Rodnitzky R, Shults C, Petsinger G, Waters C, Pfeiffer R, Biglan K, Borchert L, Montgomery A, Sutherland L, Weeks C, DeAngelis M, Sime E, Wood S, Pantella C, Harrigan M, Fussell B, Dillon S, Alexander-Brown B, Rainey P, Tennis M, Rost-Ruffner E, Brown D, Evans S, Berry D, Hall J, Shirley T, Dobson J, Fontaine D, Pfeiffer B, Brocht A, Bennett S, Daigneault S, Hodgeman K, O'Connell C, Ross T, Richard K and Watts A (2004) Pramipexole vs levodopa as initial treatment for Parkinson disease: a 4-year randomized controlled trial. Arch Neurol 61:1044-1053.
Horstink M, Tolosa E, Bonuccelli U, Deuschl G, Friedman A, Kanovsky P, Larsen JP, Lees A, Oertel W, Poewe W, Rascol O and Sampaio C (2006) Review of the therapeutic management of Parkinson's disease. Report of a joint task force of the European Federation of Neurological Societies and the Movement Disorder Society-European Section. Part I: early (uncomplicated) Parkinson's disease. Eur J Neurol 13:1170-1185.
Hwang WJ (2002) Medical Treatment of Parkinson’s Disease. Acta Neurologica Taiwanica 12:44-57.
Ishiguro N, Saito A, Yokoyama K, Morikawa M, Igarashi T and Tamai I (2005) Transport of the dopamine D2 agonist pramipexole by rat organic cation transporters OCT1 and OCT2 in kidney. Drug Metab Dispos 33:495-499.
Jonker JW and Schinkel AH (2004) Pharmacological and physiological functions of the polyspecific organic cation transporters: OCT1, 2, and 3 (SLC22A1-3). J Pharmacol Exp Ther 308:2-9.
Joyce JN (2001) Dopamine D3 receptor as a therapeutic target for antipsychotic and antiparkinsonian drugs. Pharmacol Ther 90:231-259.
Junghanns S, Glockler T and Reichmann H (2004) Switching and combining of dopamine agonists. J Neurol 251 Suppl 6:VI/19-23.
Kartzinel R, Teychenne P, Gillespie MM, Perlow M, Gielen AC, Sadowsky DA and Calne DB (1976) Bromocriptine and levodopa (with or without carbidopa) in parkinsonism. Lancet 2:272-275.
Kvernmo T, Hartter S and Burger E (2006) A review of the receptor-binding and pharmacokinetic properties of dopamine agonists. Clin Ther 28:1065-1078.
Langston JW, Ballard P, Tetrud JW and Irwin I (1983) Chronic parkinsonism in humans due to a product of meperidine-analog synthesis. Science 219:979-980.
Leabman MK and Giacomini KM (2003) Estimating the contribution of genes and environment to variation in renal drug clearance. Pharmacogenetics 13:581-584.
Lewitt PA (2008) Levodopa for the treatment of Parkinson's disease. N Engl J Med 359:2468-2476.
LeWitt PA, Boroojerdi B, MacMahon D, Patton J and Jankovic J (2007) Overnight switch from oral dopaminergic agonists to transdermal rotigotine patch in subjects with Parkinson disease. Clin Neuropharmacol 30:256-265.
Linazasoro G (2004) Conversion from dopamine agonists to pramipexole: An open-label trial in 227 patients with advanced Parkinson's disease. J Neurol 251:335-339.
Ma HI, Hwang WJ and Chen-Sea MJ (2005) Reliability and validity testing of a Chinese-translated version of the 39-item Parkinson's Disease Questionnaire (PDQ-39). Qual Life Res 14:565-569.
Maj J, Rogoz Z, Skuza G and Kolodziejczyk K (1997) Antidepressant effects of pramipexole, a novel dopamine receptor agonist. J Neural Transm 104:525-533.
Marek K, Seibyl J, Shoulson I, Holloway R, Kieburtz K, McDermott M, Kamp C, Shinaman A, Fahn S, Lang A, Weiner W and Welsh M (2002) Dopamine transporter brain imaging to assess the effects of pramipexole vs levodopa on Parkinson disease progression. JAMA 287:1653-1661.
McElvaney NG, Wilcox PG, Churg A and Fleetham JA (1988) Pleuropulmonary disease during bromocriptine treatment of Parkinson's disease. Arch Intern Med 148:2231-2236.
Miyasaki JM, Martin W, Suchowersky O, Weiner WJ and Lang AE (2002) Practice parameter: Initiation of treatment for Parkinson's disease: An evidence-based review: Report of the quality standards subcommittee of the American Academy of Neurology. Neurology 58:11-17.
Moore DJ, West AB, Dawson VL and Dawson TM (2005) Molecular pathophysiology of Parkinson's disease. Annual Review of Neuroscience 28:57-87.
Morris JC (1993) The Clinical Dementia Rating (CDR): current version and scoring rules. Neurology 43:2412-2414.
NICE (2006) Parkinson's disease diagnosis and management in primary and secondary care.http://www.nice.org.uk/nicemedia/pdf/cg035quickrefguide.pdf.
Ohno H, Nakajima M, Fujioka S, Iwamoto K and Kawamura M (2009) Overnight switching from ergot-derived dopamine agonists to pramipexole in patients with Parkinson's disease: An open preliminary trial in Japan. J Clin Neurosci 16:790-792.
Olanow CW and Obeso JA (2000a) Preventing levodopa-induced dyskinesias. Ann Neurol 47:S167-176; discussion S176-178.
Olanow CW and Obeso JA (2000b) Pulsatile stimulation of dopamine receptors and levodopa-induced motor complications in Parkinson's disease: implications for the early use of COMT inhibitors. Neurology 55:S72-77; discussion S78-81.
Olanow CW, Watts RL and Koller WC (2001) An algorithm (decision tree) for the management of Parkinson's disease (2001): Treatment guidelines. Neurology 56:S1-S88.
Pahwa R, Factor SA, Lyons KE, Ondo WG, Gronseth G, Bronte-Stewart H, Hallett M, Miyasaki J, Stevens J and Weiner WJ (2006) Practice parameter: Treatment of Parkinson disease with motor fluctuations and dyskinesia (an evidence-based review): Report of the Quality Standards Subcommittee of the American Academy of Neurology. Neurology 66:983-995.
Parkinson J (1817) An essay on the shaking palsy. London: Sherwood, Neely and Jones:1-66.
Peralta C, Wolf E, Alber H, Seppi K, Muller S, Bosch S, Wenning GK, Pachinger O and Poewe W (2006) Valvular heart disease in Parkinson's disease vs. controls: An echocardiographic study. Mov Disord 21:1109-1113.
Peto V, Jenkinson C, Fitzpatrick R and Greenhall R (1995) The development and validation of a short measure of functioning and well being for individuals with Parkinson's disease. Qual Life Res 4:241-248.
Piercey MF (1998) Pharmacology of pramipexole, a dopamine D3-preferring agonist useful in treating Parkinson's disease. Clin Neuropharmacol 21:141-151.
Poewe W (2007) Depression in Parkinson's disease. J Neurol 254:Suppl 5:49-55.
Poewe WH, Rascol O, Quinn N, Tolosa E, Oertel WH, Martignoni E, Rupp M and Boroojerdi B (2007) Efficacy of pramipexole and transdermal rotigotine in advanced Parkinson's disease: a double-blind, double-dummy, randomised controlled trial. Lancet Neurol 6:513-520.
Pogarell O, Gasser T, Van Hilten JJ, Spieker S, Pollentier S, Meier D and Oertel WH (2002) Pramipexole in patients with Parkinson's disease and marked drug resistant tremor: A randomised, double blind, placebo controlled multicentre study. J Neurol Neurosurg Psychiatry 72:713-720.
Postuma RB and Lang AE (2004) Homocysteine and levodopa: should Parkinson disease patients receive preventative therapy? Neurology 63:886-891.
Rascol O, Brooks DJ, Korczyn AD, De Deyn PP, Clarke CE and Lang AE (2000) A five-year study of the incidence of dyskinesia in patients with early Parkinson's disease who were treated with ropinirole or levodopa. N Engl J Med 342:1484-1491.
Reichmann H, Herting B, Miller A and Sommer U (2003) Switching and combining dopamine agonists. J Neural Transm 110:1393-1400.
Reichmann H, Odin P, Brecht HM, Koster J and Kraus PH (2006) Changing dopamine agonist treatment in Parkinson's disease: experiences with switching to pramipexole. J Neural Transm Suppl (71):17-25.
Relja M and Klepac N (2006) A dopamine agonist, pramipexole, and cognitive functions in Parkinson's disease. J Neurol Sci 248:251-254.
Richard IH, Maughn A and Kurlan R (1999) Do serotonin reuptake inhibitor antidepressants worsen Parkinson's disease? A retrospective case series. Mov Disord 14:155-157.
Rothman RB, Baumann MH, Savage JE, Rauser L, McBride A, Hufeisen SJ and Roth BL (2000) Evidence for possible involvement of 5-HT(2B) receptors in the cardiac valvulopathy associated with fenfluramine and other serotonergic medications. Circulation 102:2836-2841.
Sautel F, Griffon N, Levesque D, Pilon C, Schwartz JC and Sokoloff P (1995) A functional test identifies dopamine agonists selective for D3 versus D2 receptors. NeuroReport 6:329-332.
Schapira AHV (2007) Treatment options in the modern management of Parkinson disease. Arch Neurol 64:1083-1088.
Schrag A (2005) Entacapone in the treatment of Parkinson's disease. Lancet Neurol 4:366-370.
Sethi K (2008) Levodopa unresponsive symptoms in Parkinson disease. Mov Disord 23:S521–S533.
Sibley DR and Monsma FJ, Jr. (1992) Molecular biology of dopamine receptors. Trends Pharmacol Sci 13:61-69.
Simuni T, Lyons KE, Pahwa R, Hauser RA, Comella C, Elmer L and Weintraub D (2009) Treatment of Early Parkinson's Disease. Part 1. Eur Neurol 61:193-205.
Simuni T and Sethi K (2008) Nonmotor manifestations of Parkinson's disease. Ann Neurol 64 Suppl 2:S65-80.
Spillantini MG, Crowther RA, Jakes R, Hasegawa M and Goedert M (1998) α-Synuclein in filamentous inclusions of Lewy bodies from Parkinson's disease and dementia with Lewy bodies. Proc Natl Acad Sci U S A 95:6469-6473.
Stewart D, Morgan E, Burn D, Grosset D, Chaudhuri KR, MacMahon D, Needleman F, Macphee G and Heywood P (2004) Dopamine agonist switching in Parkinson's disease. Hosp Med 65:215-219.
Suchowersky O, Gronseth G, Perlmutter J, Reich S, Zesiewicz T and Weiner WJ (2006) Practice parameter: Neuroprotective strategies and alternative therapies for Parkinson disease (an evidence-based review): Report of the Quality Standards Subcommittee of the American Academy of Neurology. Neurology 66:976-982.
Takahashi H, Nogawa S, Tachibana H, Kawamura J, Abe T, Ogino Y, Kashihara K, Hamada T and Kowa H (2008) Pramipexole safely replaces ergot dopamine agonists with either rapid or slow switching. J Int Med Res 36:106-114.
Tanner CM and Goldman SM (1996) Epidemiology of Parkinson's disease. Neurol Clin 14:317-335.
Thobois S (2006) Proposed dose equivalence for rapid switch between dopamine receptor agonists in Parkinson's disease: a review of the literature. Clin Ther 28:1-12.
Thobois S, Delamarre-Damier F and Derkinderen P (2005) Treatment of motor dysfunction in Parkinson's disease: an overview. Clin Neurol Neurosurg 107:269-281.
Van Camp G, Flamez A, Cosyns B, Weytjens C, Muyldermans L, Van Zandijcke M, De Sutter J, Santens P, Decoodt P, Moerman C and Schoors D (2004) Treatment of Parkinson's disease with pergolide and relation to restrictive valvular heart disease. Lancet 363:1179-1183.
Van Den Eeden SK, Tanner CM, Bernstein AL, Fross RD, Leimpeter A, Bloch DA and Nelson LM (2003) Incidence of Parkinson's disease: variation by age, gender, and race/ethnicity. Am J Epidemiol 157:1015-1022.
Visser M, Leentjens AF, Marinus J, Stiggelbout AM and van Hilten JJ (2006) Reliability and validity of the Beck depression inventory in patients with Parkinson's disease. Mov Disord 21:668-672.
WHO (2009) ATC/DDD Index 2009. http://www.whocc.no/atcddd/indexdatabase/index.php?query=N04BC.
Whone AL, Watts RL, Stoessl AJ, Davis M, Reske S, Nahmias C, Lang AE, Rascol O, Ribeiro MJ, Remy P, Poewe WH, Hauser RA and Brooks DJ (2003) Slower progression of Parkinson's disease with ropinirole versus levodopa: The REAL-PET study. Ann Neurol 54:93-101.
Wong KS, Lu CS, Shan DE, Yang CC, Tsoi TH and Mok V (2003) Efficacy, safety, and tolerability of pramipexole in untreated and levodopa-treated patients with Parkinson's disease. J Neurol Sci 216:81-87.
Wright CE, Sisson TL, Ichhpurani AK and Peters GR (1997) Steady-state pharmacokinetic properties of pramipexole in healthy volunteers. J Clin Pharmacol 37:520-525.
Yamamoto M and Uesugi T (2007) Dopamine agonists and valvular heart disease in patients with Parkinson's disease: Evidence and mystery. J Neurol 254:74-78.
Zanettini R, Antonini A, Gatto G, Gentile R, Tesei S and Pezzoli G (2007) Valvular heart disease and the use of dopamine agonists for Parkinson's disease. N Engl J Med 356:39-46.
中央健康保險局 (2009) 健保用藥品項查詢. http://www.nhi.gov.tw/inquire/query1.asp?menu=1&menu_id=8&WD_ID=8.
行政院衛生署 (2005) 全民健康保險醫療統計年報. http://www.doh.gov.tw/statistic/data/全民健康保險統計年報/94醫療統計年報/94全民健康保險醫療統計年報.rar.
行政院衛生署 (2006) 全民健康保險醫療統計年報. http://www.doh.gov.tw/statistic/data/全民健康保險統計年報/95醫療統計年報/pdf.pdf.

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系統識別號 U0007-0307200913545800
論文名稱(中文) 薑黃素及其結構類似物對於人類纖維肉瘤細胞 (HT-1080)抑制轉移活性之相關研究
論文名稱(英文) Studies on Curcumin and its analogs on anti-metastatic activities of human fibrosarcoma cells (HT-1080)
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 97
學期 2
出版年 98
研究生(中文) 李卓涵
學號 M303096001
學位類別 碩士
語文別 中文
口試日期 2009-06-05
論文頁數 83頁
口試委員 委員-林耀輝
委員-傅淑玲
委員-林時宜
委員-梁有志
指導教授-侯文琪
關鍵字(中) 薑黃素
金屬基質蛋白分解酶
抗癌症轉移
關鍵字(英) curcumin
matrix metalloproteinase
antimetastasis
學科別分類
中文摘要 薑黃素 (curcumin)為存在於植物薑黃中的黃色色素,已被研究出具有抗發炎、抗癌症侵襲以及抗轉移等多種活性。目前已有相當多研究試圖改變薑黃素的化學結構以合成生物活性優於薑黃素的類似物。根據文獻,金屬基質蛋白酵素 (Matrix metalloproteinases, MMPs)以及尿激酶型纖溶酶原激活劑 (urokinase-type plasminogen activator, uPA)在癌細胞進行轉移過程對於細胞外間質的水解扮演關鍵的角色。本篇研究使用薑黃素及其二十三個結構類似物對於具有高度轉移活性的人類纖維肉瘤細胞 (HT-1080)進行MMP-9活性、蛋白質及基因表現抑制活性篩選,並探討其可能機制。除了二十四號化合物和三十一號化合物以2.5 μM處理細胞之外,薑黃素及其餘結構類似物以濃度5 μM處理癌細胞並無顯著細胞毒性。由明膠酵素電泳法結果顯示,二號化合物 (兩側苯環含有六個甲氧基取代)、七號化合物 (兩側苯環含有三個氫氧基、一個甲氧基)與薑黃素在同濃度5 μM比較下具有較強抑制MMP-9活性的能力。本研究也同時以濃度2.5 μM的薑黃素、二號、七號、二十四號 (兩側苯環含有三個氫氧基取代)及三十一號化合物 (兩側苯環含有兩個氫氧基,兩個甲氧基取代)處理HT-1080細胞,比較以上結構類似物對於MMP-9表現的影響。經由西方墨點法、反轉錄聚合酶連鎖反應結果顯示,二號、七號、二十四號及三十一號化合物抑制MMP-9基因的表現;由明膠酵素電泳法、纖維蛋白酵素電泳法及MMP-9酵素活性試驗觀察到四個化合物能抑制HT-1080細胞的MMP-9活性,進而抑制了HT-1080細胞的遷移及侵襲能力。綜合以上,二號化合物和七號化合物比薑黃素、二十四號化合物和三十一號化合物具有更好的抗轉移活性。第一型血紅素加氧酶 (Heme oxygenase 1, HO-1),為一種遭遇壓力便會誘導產生的蛋白質,且HO-1的誘導與ROS產生具有關連性,有研究指出其可能為抑制MMPs的原因。以西方墨點法觀察薑黃素、二號、七號、二十四號及三十一號化合物對於HT-1080細胞內的HO-1蛋白表現量,結果顯示薑黃素,二號化合物及三十一號化合物可以顯著誘導HO-1蛋白質的表現,然而七號化合物及二十四號化合物卻無法誘導之。以上結果顯示,薑黃素、二號、七號、二十四號及三十一號化合物於體外試驗觀察到其對於癌細胞株遷移、侵襲能力的抑制機轉可能來自於抑制MMP-9基因表現和抑制uPA活性。另外,二號化合物抑制MMP-9活性的能力較佳推測有可能是其可顯著誘導細胞大量產生HO-1以對抗PMA所誘導產生的ROS。由以上結果得知二號、七號、二十四號及三十一號化合物分別呈現不同程度抑制MMP-9及uPA且活性皆優於薑黃素。
英文摘要 Curcumin, a yellowish pigment existed in turmeric plant has been shown to exhibit numerous biological properties, such as anti-inflammation, antioxidation, anti-invasion, and antimetastatic activity. Chemical modifications of curcumin have been studied intensively in an attempt to find an analog with similar but enhanced biological activities of curcumin. In literatures, matrix metalloproteinases (MMPs) and urokinase plasminogen activator (uPA) played important roles in cancer cell invasion by hydrolysing extracellular matrix (ECM). In this study, curcumin and 23 novel analogs were used to screen for inhibition of matrix metalloproteinase-9 (MMP-9) activities, protein and gene expression of highly metastatic HT-1080 human fibrosarcoma cells to explore the mechanisms of action. All tested compounds, except analog 24 and analog 31 at concentration of 2.5 μM showed no cytotoxic activities at 5 μM. The analog 2 (six methoxy substitutes in phenyl groups) and analog 7 (three hydroxyl and one methoxyl substitutes in phenyl groups) showed higher MMP-9 inhibitory activities than curcumin did at 5 μM in gelatin zymography analysis. We comparatively examined the influence of analog 2, 7, 24 (three hydroxyl substitutes in phenyl groups), and 31(two hydroxyl and two methoxyl substitutes in phenyl groups) on the expression of MMP-9 of HT-1080 cells at concentration of 2.5 μM. Analog 2, 7, 24 and 31 suppressed the gene expression of MMP-9 as the results of RT-PCR assay and Western blot assay, and decreased their corresponding activities in HT-1080 cells revealed by gelatin zymography assay, MMP-9 activity ELISA and fibrin zymography assay which resulted in inhibition of HT-1080 cell invasion and migration differentially. All in all, analog 2 and 7 showed higher anti-metastatic activities than analog 24 and analog 31 did. Heme oxygenase 1(HO-1), a stress-responsive enzyme, which was found to correlate with production of reactive oxygen species (ROS), suggesting a causative relationship on MMP inhibition. Therefore, the effect of curcumin and analog 2, 7, 24 and 31 on HO-1 protein expression were also studied. Interestingly, exposure to curcumin, analog 2 and 31 treatment maximally induce HO-1 protein expression in HT-1080 cells, however, analog 7 and 24 were less apparently. These data suggested that the inhibitory effects of curcumin and analog 2, 7, 24 and 31 on MMP-9 gene expression and uPA activity was closely related to tumor invasion and migration in vitro. Furthermore, analog 2 showed highly MMP-9 inhibitory activity which possibly involved mechanisms related to its ability to induce HO-1 to suppress PMA-induced ROS, which can activate MMP-9 gene expression. In summary, these data demonstrated that analog 2, 7, 24 and 31 show higher anti-metastasis potency than curcumin by the differentially down-regulation of MMP-9 and uPA.
論文目次 目錄
目錄 I
圖目錄 III
表目錄 VII
中文摘要 VIII
Abstract X
縮寫表 XII

第一章 緒論........ 1
第一節 前言........ 1
第二節 文獻回顧.... 2
I. 薑黃素 2
II. 癌症轉移........9
III. 血管新生 (Angiogenesis)..10
IV. 金屬基質蛋白酵素..........11
V. 纖維蛋白溶解酶系統 (Plasminogen activator/plasmin system) 19
VI. 血紅素加氧酶 (Heme oxygenase)...... 21
第二章 材料與方法.. 23
第一節 實驗材料.....23
I. 材料與儀器...... 23
II. 細胞株 (HT-1080)之處理... 27
第二節 實驗方法.....30
第三章 結果........ 37
I. 薑黃素及其二十三個結構類似物對於人類纖維肉瘤細胞 (HT-1080) 的MMP-9活性及其對細胞毒性的影響...37
II. 薑黃素及其結構類似物(二號、七號、二十四號及三十一號化合物)對於HT-1080細胞株之MMP-9活性的抑制實驗........ 40
III. 利用酵素連結免疫吸附法 (ELISA)判定薑黃素、薑黃素結構類似物(二號、七號、二十四號及三十一號)對於HT-1080之MMP-9酵素活性抑制實驗..........44
IV. 薑黃素及其薑黃素類似物結構類似物(二號、七號、二十四號及三十一號化合物)對於MMP-9酵素活性抑制實驗......... 46
V. 薑黃素及其薑黃素類似物結構類似物(二號、七號、二十四號及三十一號化合物)對於HT-1080之MMP-9蛋白表現抑制實驗...48
VI. 薑黃素及其結構類似物(二號、七號、二十四號及三十一號化合物)對於HT-1080之MMP-9基因表現的抑制實驗 51
VII. 薑黃素及其結構類似物(二號、七號、二十四號及三十一號化合物)對於HT-1080 uPA之活性抑制試驗....... 54
VIII. 薑黃素、薑黃素類似物(二號、七號、二十四號及三十一號化合物)對於HT-1080遷移與侵襲的抑制試驗....57
IX. 薑黃素及其結構類似物(二號、七號、二十四號及三十一號化合物)對於HO-1蛋白質表現量的影響 63
第四章 討論........ 66
第五章 結論........ 73
第六章 參考文獻.....75

圖目錄
附圖 一、類薑黃素的化學結構式 3
附圖 二、薑黃素在人體內的代謝產物.......3
附圖 三、薑黃素影響癌症生成的階段及相關因子.......5
附圖 四、薑黃素影響的細胞內訊息傳遞路徑 6
附圖 五、薑黃素具有潛力預防及治療的癌症 6
附圖 六、薑黃素可改變的化學基團........ 7
附圖 七、癌症轉移過程圖.......10
附圖 八、調控MMPs表現與活性的階層.......13
附圖 九、MMPs的結構圖........ 14
附圖 十、pro-MMPs的活化...... 15
附圖 十一、纖維蛋白溶解酶系統相關機轉...20
附圖 十二、誘導HO-1表現的因子 21
附圖 十三、血基質分解途徑.... 22

圖 一、薑黃素及二十三個結構類似物在濃度5 μM下處理HT-1080 細胞株,24小時之後細胞存活情形 (其中二十四號及三十一號化合物為 2.5 μM)。....... 38
圖 二、薑黃素及二十三個結構類似物在濃度5 μM下處理HT-1080 細胞株24小時,利用明膠-SDS PAGE膠體判定其對MMP-9活性的影響(其中二十四號及三十一號化合物為2.5 μM)。...39
圖 三、 不同濃度的薑黃素、二號及七號化合物處理HT-1080細胞株, 24小時後之細胞存活率。..........41
圖 四、薑黃素及二號、七號化合物處理HT-1080 細胞株24小時,利用明膠-SDS PAGE膠體判定其對MMP-9活性的影響。......42
圖 五、薑黃素及結構類似物(二號、七號、二十四號及三十一號化合物)處理HT-1080 細胞株24小時,利用明膠-SDS PAGE膠體判定其對MMP-9活性的影響。...43
圖 六、利用酵素連結免疫吸附法以不同濃度的薑黃素及結構類似物 (二號、七號、二十四號及三十一號化合物)處理HT-1080細胞株24小時,對其MMP-9酵素活性的影響。 45
圖 七、薑黃素及其薑黃素類似物結構類似物(二號、七號、二十四號及三十一號化合物)對於MMP-9酵素活性的影響。....... 47
圖 八、 薑黃素、二號、七號化合物處理HT-1080細胞株24小時,對其細胞分泌MMP-9蛋白質量的影響。.........49
圖 九、薑黃素、二號、七號、二十四號及三十一號化合物處理HT-1080細胞株24小時,對其細胞分泌MMP-9蛋白質量的影響。.....50
圖 十、薑黃素、二號、七號化合物處理HT-1080細胞株三小時後,其對MMP-9 mRNA表現的影響....... 52
圖 十一、 薑黃素、二號、七號、二十四號及三十一號化合物處理HT-1080細胞株三小時後,其對MMP-9 mRNA表現的影響.. 53
圖 十二、薑黃素及二號、七號化合物處理HT-1080 細胞株24小時,利用纖維蛋白-SDS PAGE膠體判定其對uPA活性的影響。..55
圖 十三、薑黃素及二號、七號、二十四號及三十一號化合物處理HT-1080 細胞株24小時,利用纖維蛋白-SDS PAGE膠體判定其對uPA活性的影響。..56
圖 十四、濃度5 μM的薑黃素、二號及七號化合物對於HT-1080細胞株遷移的影響。..... 59
圖 十五、薑黃素,結構類似物二號、七號、二十四號及三十一號化合物對於HT-1080細胞株遷移的影響。...... 60
圖 十六、薑黃素、二號化合物以及七號化合物對於HT-1080細胞侵襲的影響。..61
圖 十七、薑黃素、二號、七號、二十四號及三十一號化合物對於HT-1080細胞侵襲的影響。..........62
圖 十八、薑黃素、二號、七號化合物對於細胞株HT-1080的HO-1蛋白質表現量的影響。....64
圖 十九、薑黃素、二號、七號、二十四號及三十一號化合物對於HT-1080細胞HO-1蛋白質表現量的影響。........65

參考文獻 Aggarwal BB, Kumar A, Bharti AC (2003) Anticancer potential of curcumin: preclinical and clinical studies. Anticancer Res 23: 363-98

Aggarwal BB, Sundaram C, Malani N, Ichikawa H (2007) Curcumin: the Indian solid gold. Adv Exp Med Biol 595: 1-75

Ahmad A, Hart IR (1997) Mechanisms of metastasis. Crit Rev Oncol Hematol 26: 163-73

Albini A, Iwamoto Y, Kleinman HK, Martin GR, Aaronson SA, Kozlowski JM, McEwan RN (1987) A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res 47: 3239-45

Anand P, Sundaram C, Jhurani S, Kunnumakkara AB, Aggarwal BB (2008a) Curcumin and cancer: an "old-age" disease with an "age-old" solution. Cancer Lett 267: 133-64

Anand P, Thomas SG, Kunnumakkara AB, Sundaram C, Harikumar KB, Sung B, Tharakan ST, Misra K, Priyadarsini IK, Rajasekharan KN, Aggarwal BB (2008b) Biological activities of curcumin and its analogues (Congeners) made by man and Mother Nature. Biochem Pharmacol

Anand P, Thomas SG, Kunnumakkara AB, Sundaram C, Harikumar KB, Sung B, Tharakan ST, Misra KP, Priyadarsini IK, Rajasekharan KN, Aggarwal BB (2008c) Biological activities of curcumin and its analogues (Congeners) made by man and Mother Nature. Biochem Pharmacol

Andreasen PA, Kjoller L, Christensen L, Duffy MJ (1997) The urokinase-type plasminogen activator system in cancer metastasis: a review. Int J Cancer 72: 1-22

Bernhard EJ, Muschel RJ, Hughes EN (1990) Mr 92,000 gelatinase release correlates with the metastatic phenotype in transformed rat embryo cells. Cancer Res 50: 3872-7

Bjorklund M, Koivunen E (2005) Gelatinase-mediated migration and invasion of cancer cells. Biochim Biophys Acta 1755: 37-69

Brooks SA, Lomax-Browne HJ, Carter TM, Kinch CE, Hall DM (2009) Molecular interactions in cancer cell metastasis. Acta Histochem

Carmeliet P, Jain RK (2000) Angiogenesis in cancer and other diseases. Nature 407: 249-57

Carmeliet P, Moons L, Lijnen R, Baes M, Lemaitre V, Tipping P, Drew A, Eeckhout Y, Shapiro S, Lupu F, Collen D (1997) Urokinase-generated plasmin activates matrix metalloproteinases during aneurysm formation. Nat Genet 17: 439-44

Chang C, Werb Z (2001) The many faces of metalloproteases: cell growth, invasion, angiogenesis and metastasis. Trends Cell Biol 11: S37-43

Chen WF, Deng SL, Zhou B, Yang L, Liu ZL (2006) Curcumin and its analogues as potent inhibitors of low density lipoprotein oxidation: H-atom abstraction from the phenolic groups and possible involvement of the 4-hydroxy-3-methoxyphenyl groups. Free Radic Biol Med 40: 526-35

Coussens LM, Fingleton B, Matrisian LM (2002) Matrix metalloproteinase inhibitors and cancer: trials and tribulations. Science 295: 2387-92

Dai F, Chen WF, Zhou B, Yang L, Liu ZL (2009) Antioxidative effects of curcumin and its analogues against the free-radical-induced peroxidation of linoleic acid in micelles. Phytother Res

Deryugina EI, Quigley JP (2006) Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev 25: 9-34

Duffy MJ (2004) The urokinase plasminogen activator system: role in malignancy. Curr Pharm Des 10: 39-49

Duffy MJ, Duggan C (2004) The urokinase plasminogen activator system: a rich source of tumour markers for the individualised management of patients with cancer. Clin Biochem 37: 541-8

Egeblad M, Werb Z (2002) New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer 2: 161-74

Engelse MA, Hanemaaijer R, Koolwijk P, van Hinsbergh VW (2004a) The fibrinolytic system and matrix metalloproteinases in angiogenesis and tumor progression. Semin Thromb Hemost 30: 71-82

Engelse MA, Hanemaaijer R, Koolwijk P, van Hinsbergh VWM (2004b) The fibrinolytic system and matrix metalloproteinases in angiogenesis and tumor progression. Seminars in Thrombosis and Hemostasis 30: 71-82

Farombi EO, Surh YJ (2006) Heme oxygenase-1 as a potential therapeutic target for hepatoprotection. J Biochem Mol Biol 39: 479-91

Folgueras AR, Pendas AM, Sanchez LM, Lopez-Otin C (2004) Matrix metalloproteinases in cancer: from new functions to improved inhibition strategies. International Journal of Developmental Biology 48: 411-424

Fu X, Parks WC, Heinecke JW (2008) Activation and silencing of matrix metalloproteinases. Semin Cell Dev Biol 19: 2-13

Goel A, Kunnumakkara AB, Aggarwal BB (2008) Curcumin as "Curecumin": from kitchen to clinic. Biochem Pharmacol 75: 787-809

Gross J, Lapiere CM (1962) Collagenolytic activity in amphibian tissues: a tissue culture assay. Proc Natl Acad Sci U S A 48: 1014-22

Hahm ER, Gho YS, Park S, Park C, Kim KW, Yang CH (2004) Synthetic curcumin analogs inhibit activator protein-1 transcription and tumor-induced angiogenesis. Biochem Biophys Res Commun 321: 337-44

Handsley MM, Edwards DR (2005) Metalloproteinases and their inhibitors in tumor angiogenesis. Int J Cancer 115: 849-60

Hatcher H, Planalp R, Cho J, Torti FM, Torti SV (2008) Curcumin: from ancient medicine to current clinical trials. Cell Mol Life Sci 65: 1631-52

Huang MT, Ma W, Lu YP, Chang RL, Fisher C, Manchand PS, Newmark HL, Conney AH (1995) Effects of curcumin, demethoxycurcumin, bisdemethoxycurcumin and tetrahydrocurcumin on 12-O-tetradecanoylphorbol-13-acetate-induced tumor promotion. Carcinogenesis 16: 2493-7

Itokawa H, Shi Q, Akiyama T, Morris-Natschke SL, Lee KH (2008) Recent advances in the investigation of curcuminoids. Chin Med 3: 11

Jeong GS, Oh GS, Pae HO, Jeong SO, Kim YC, Shin MK, Seo BY, Han SY, Lee HS, Jeong JG, Koh JS, Chung HT (2006) Comparative effects of curcuminoids on endothelial heme oxygenase-1 expression: ortho-methoxy groups are essential to enhance heme oxygenase activity and protection. Exp Mol Med 38: 393-400

Jiang Y, Goldberg ID, Shi YE (2002) Complex roles of tissue inhibitors of metalloproteinases in cancer. Oncogene 21: 2245-52

John A, Tuszynski G (2001) The role of matrix metalloproteinases in tumor angiogenesis and tumor metastasis. Pathol Oncol Res 7: 14-23

Johnson LL, Dyer R, Hupe DJ (1998) Matrix metalloproteinases. Curr Opin Chem Biol 2: 466-71

Khanna C, Hunter K (2005a) Modeling metastasis in vivo. Carcinogenesis 26: 513-523

Khanna C, Hunter K (2005b) Modeling metastasis in vivo. Carcinogenesis 26: 513-23

Khasigov PZ, Podobed OV, Gracheva TS, Salbiev KD, Grachev SV, Berezov TT (2003) Role of matrix metalloproteinases and their inhibitors in tumor invasion and metastasis. Biochemistry (Mosc) 68: 711-7

Kikuchi G, Yoshida T, Noguchi M (2005) Heme oxygenase and heme degradation. Biochemical and Biophysical Research Communications 338: 558-567

Kleiner DE, Stetler-Stevenson WG (1994) Quantitative zymography: detection of picogram quantities of gelatinases. Anal Biochem 218: 325-9

Konstantinopoulos PA, Karamouzis MV, Papatsoris AG, Papavassiliou AG (2008) Matrix metalloproteinase inhibitors as anticancer agents. Int J Biochem Cell Biol 40: 1156-68

Kunnumakkara AB, Anand P, Aggarwal BB (2008) Curcumin inhibits proliferation, invasion, angiogenesis and metastasis of different cancers through interaction with multiple cell signaling proteins. Cancer Lett

Lee M, Fridman R, Mobashery S (2004) Extracellular proteases as targets for treatment of cancer metastases. Chemical Society Reviews 33: 401-409

Lee SO, Jeong YJ, Kim M, Kim CH, Lee IS (2008) Suppression of PMA-induced tumor cell invasion by capillarisin via the inhibition of NF-kappaB-dependent MMP-9 expression. Biochem Biophys Res Commun 366: 1019-24

Lee SO, Jeong YJ, Yu MH, Lee JW, Hwangbo MH, Kim CH, Lee IS (2006) Wogonin suppresses TNF-alpha-induced MMP-9 expression by blocking the NF-kappaB activation via MAPK signaling pathways in human aortic smooth muscle cells. Biochem Biophys Res Commun 351: 118-25

Li WY, Chong SS, Huang EY, Tuan TL (2003) Plasminogen activator/plasmin system: a major player in wound healing? Wound Repair Regen 11: 239-47

Lin CW, Shen SC, Hou WC, Yang LY, Chen YC (2008) Heme oxygenase-1 inhibits breast cancer invasion via suppressing the expression of matrix metalloproteinase-9. Mol Cancer Ther 7: 1195-206

Lin CW, Yang LY, Shen SC, Chen YC (2007) IGF-I plus E2 induces proliferation via activation of ROS-dependent ERKs and JNKs in human breast carcinoma cells. J Cell Physiol 212: 666-74

Liotta LA, Kleinerman J, Catanzaro P, Rynbrandt D (1977) Degradation of basement membrane by murine tumor cells. J Natl Cancer Inst 58: 1427-31

Liotta LA, Tryggvason K, Garbisa S, Hart I, Foltz CM, Shafie S (1980) Metastatic potential correlates with enzymatic degradation of basement membrane collagen. Nature 284: 67-8

Lopez-Lazaro M (2008) Anticancer and carcinogenic properties of curcumin: Considerations for its clinical development as a cancer chemopreventive and chemotherapeutic agent. Mol Nutr Food Res

Mazumder A, Neamati N, Sunder S, Schulz J, Pertz H, Eich E, Pommier Y (1997) Curcumin analogs with altered potencies against HIV-1 integrase as probes for biochemical mechanisms of drug action. J Med Chem 40: 3057-63

McNally SJ, Harrison EM, Ross JA, Garden OJ, Wigmore SJ (2006) Curcumin induces heme oxygenase-1 in hepatocytes and is protective in simulated cold preservation and warm reperfusion injury. Transplantation 81: 623-6

Menon LG, Kuttan R, Kuttan G (1995) Inhibition of lung metastasis in mice induced by B16F10 melanoma cells by polyphenolic compounds. Cancer Lett 95: 221-5

Menon LG, Kuttan R, Kuttan G (1999) Anti-metastatic activity of curcumin and catechin. Cancer Lett 141: 159-65

Mohan R, Sivak J, Ashton P, Russo LA, Pham BQ, Kasahara N, Raizman MB, Fini ME (2000) Curcuminoids inhibit the angiogenic response stimulated by fibroblast growth factor-2, including expression of matrix metalloproteinase gelatinase B. J Biol Chem 275: 10405-12

Morse D, Choi AMK (2005) Heme oxygenase-1 - From bench to bedside. American Journal of Respiratory and Critical Care Medicine 172: 660-670

Motterlini R, Foresti R, Bassi R, Green CJ (2000) Curcumin, an antioxidant and anti-inflammatory agent, induces heme oxygenase-1 and protects endothelial cells against oxidative stress. Free Radic Biol Med 28: 1303-12

Nack-Shick Choi K-SY, Jin-Young Lee, Kyoung-Yoen Han and Seung-Ho Kim (2001) Comparison of Three Substrates (Casein, Fibrin, and Gelatin) in Zymographic Gel. Journal of Biochemistry and Molecular Biology 34: 531-536

Nagase H (1997) Activation mechanisms of matrix metalloproteinases. Biol Chem 378: 151-60

Nociari MM, Shalev A, Benias P, Russo C (1998) A novel one-step, highly sensitive fluorometric assay to evaluate cell-mediated cytotoxicity. J Immunol Methods 213: 157-67

Osawa T, Sugiyama Y, Inayoshi M, Kawakishi S (1995) Antioxidative activity of tetrahydrocurcuminoids. Biosci Biotechnol Biochem 59: 1609-12

Otterbein LE, Soares MP, Yamashita K, Bach FH (2003) Heme oxygenase-1: unleashing the protective properties of heme. Trends in Immunology 24: 449-455

Overall CM, Lopez-Otin C (2002) Strategies for MMP inhibition in cancer: innovations for the post-trial era. Nat Rev Cancer 2: 657-72

Polette M, Nawrocki-Raby B, Gilles C, Clavel C, Birembaut P (2004) Tumour invasion and matrix metalloproteinases. Crit Rev Oncol Hematol 49: 179-86

Ra HJ, Parks WC (2007) Control of matrix metalloproteinase catalytic activity. Matrix Biol 26: 587-96

Ruby AJ, Kuttan G, Babu KD, Rajasekharan KN, Kuttan R (1995) Anti-tumour and antioxidant activity of natural curcuminoids. Cancer Lett 94: 79-83

Rundhaug JE (2005) Matrix metalloproteinases and angiogenesis. J Cell Mol Med 9: 267-85

Salo T, Liotta LA, Keski-Oja J, Turpeenniemi-Hujanen T, Tryggvason K (1982) Secretion of basement membrane collagen degrading enzyme and plasminogen activator by transformed cells--role in metastasis. Int J Cancer 30: 669-73

Sandur SK, Pandey MK, Sung B, Ahn KS, Murakami A, Sethi G, Limtrakul P, Badmaev V, Aggarwal BB (2007) Curcumin, demethoxycurcumin, bisdemethoxycurcumin, tetrahydrocurcumin and turmerones differentially regulate anti-inflammatory and anti-proliferative responses through a ROS-independent mechanism. Carcinogenesis 28: 1765-73

Sharma RA, Gescher AJ, Steward WP (2005) Curcumin: the story so far. Eur J Cancer 41: 1955-68

Simon A, Allais DP, Duroux JL, Basly JP, Durand-Fontanier S, Delage C (1998) Inhibitory effect of curcuminoids on MCF-7 cell proliferation and structure-activity relationships. Cancer Lett 129: 111-6

Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150: 76-85

Stetler-Stevenson WG (1999) Matrix metalloproteinases in angiogenesis: a moving target for therapeutic intervention. J Clin Invest 103: 1237-41

Stetler-Stevenson WG, Hewitt R, Corcoran M (1996) Matrix metalloproteinases and tumor invasion: from correlation and causality to the clinic. Semin Cancer Biol 7: 147-54

Steward WP, Gescher AJ (2008) Curcumin in cancer management: Recent results of analogue design and clinical studies and desirable future research. Mol Nutr Food Res 52: 1005-1009

Vassalli JD, Sappino AP, Belin D (1991) The plasminogen activator/plasmin system. J Clin Invest 88: 1067-72

Venkateswarlu S, Ramachandra MS, Subbaraju GV (2005) Synthesis and biological evaluation of polyhydroxycurcuminoids. Bioorg Med Chem 13: 6374-80

Visse R, Nagase H (2003) Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry. Circ Res 92: 827-39

Wei QY, Chen WF, Zhou B, Yang L, Liu ZL (2006) Inhibition of lipid peroxidation and protein oxidation in rat liver mitochondria by curcumin and its analogues. Biochim Biophys Acta 1760: 70-7

Woessner JF, Jr. (1991) Matrix metalloproteinases and their inhibitors in connective tissue remodeling. FASEB J 5: 2145-54

Wu WS, Tsai RK, Chang CH, Wang S, Wu JR, Chang YX (2006) Reactive oxygen species mediated sustained activation of protein kinase C alpha and extracellular signal-regulated kinase for migration of human hepatoma cell Hepg2. Mol Cancer Res 4: 747-58

Yodkeeree S, Chaiwangyen W, Garbisa S, Limtrakul P (2008a) Curcumin, demethoxycurcumin and bisdemethoxycurcumin differentially inhibit cancer cell invasion through the down-regulation of MMPs and uPA. J Nutr Biochem

Yodkeeree S, Garbisa S, Limtrakul P (2008b) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA. Acta Pharmacol Sin 29: 853-60

Yoon SO, Park SJ, Yun CH, Chung AS (2003) Roles of matrix metalloproteinases in tumor metastasis and angiogenesis. J Biochem Mol Biol 36: 128-37

Zhao Y, Lyons CE, Jr., Xiao A, Templeton DJ, Sang QA, Brew K, Hussaini IM (2008) Urokinase directly activates matrix metalloproteinases-9: a potential role in glioblastoma invasion. Biochem Biophys Res Commun 369: 1215-20




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系統識別號 U0007-0608200921174400
論文名稱(中文) 評估乳癌病患使用G-CSF預防化學治療期間嗜中性白血球低下及併發性發燒之研究
論文名稱(英文) Study on the Use of G-CSF in Breast Cancer Patients for Managing Chemotherapy-induced Febrile Neutropenia
校院名稱 臺北醫學大學
系所名稱(中) 藥學研究所
系所名稱(英) Graduate Institute of Pharmacy
學年度 97
學期 2
出版年 98
研究生(中文) 劉承忠
學號 M301096034
學位類別 碩士
語文別 英文
口試日期 2009-06-24
論文頁數 61頁
口試委員 指導教授-許秀蘊
共同指導教授-闕壯卿
共同指導教授-姜紹青
委員-陳國棟
委員-陳寶年
委員-吳姿樺
關鍵字(中) 嗜中性白血球低下合併發燒(FN);顆粒性白血球群落刺激因子(G-CSF);乳癌;初級預防;次級預防;輔助性化學治療;Adriamycin
Paclitaxel
Cyclophosphamide
5-fluorouracil.
關鍵字(英) Febrile neutropenia (FN)
Granulocyte colony-stimulating factor (G-CSF)
Breast cancer
Primary prophylaxis
Secondary prophylaxis
Adjuvant chemotherapy
Adriamycin
Paclitaxel
Cyclophosphamide
5-fluorouracil.
學科別分類
中文摘要 動機 嗜中性白血球低下合併發燒 (FN) 是病患在接受癌症化療時產生的嚴重血液副作用,臨床上可使用抗生素或顆粒性白血球群落刺激因子(G-CSF)來預防。G-CSF為細胞激素 (造血生長刺激因子),它能調節造血幹細胞的增殖,分化和功能,並呈現劑量依賴性的增加血液循環中嗜中性白血球之數量。在早期的臨床試驗中顯示,於小細胞肺癌病患中,以G-CSF來做初級預防性治療可縮短化療引起嗜中性白血球低下的時間,並進一步降低FN、感染、住院及使用抗生素約50 %之發生率。在此研究中,G-CSF自化療後第四天開始給予直到第十七天,FN之發生率從77%降到40%。而另一個針對乳癌病患的研究中,在經過化療後第四天至第十天的G-CSF給予後,對於接受TAC (adriamycin, paclitaxel, 及cyclophosphamide)化療的病患,可使FN的發生率從27.5%降至7.5%。根據目前臨床準則的建議,G-CSF只適用於FN風險大於20%的病患 。

目的 G-CSF屬於高價位的藥品,目前在辜公亮和信治癌中心,G-CSF由於健保局的規範下通常開予三天的處方。本研究的主要目標為研究在乳癌病患在接受G-CSF初級及次級預防後其FN的發生率;次要目標為探討G-CSF的處方型態和評估在目前健保的給付規範下G-CSF使用的成本效益分析。

研究設計 本研究為病歷回溯性研究,納入2007年於和信治癌中心接受化學治療或lenograstim的乳癌病患。收集的資料包括了病人的年紀、性別、腫瘤類型、化療處方及劑量、G-CSF使用劑量及期間、住院時間及成本等。接著,依照病患接受的化療處方其FN之風險分成三組,分別為高(>20%)、中(10-20%)、低(<10%),以找出在不同組之FN風險及其G-CSF使用的型態。此外,我們進一步著重在研究乳癌病患接受化療處方TAC, ATC 和 CAF其使用G-CSF的型態及FN之發生率,並和目前文獻研究的結果加以比較以評估臨床療效及延長或增加G-CSF使用時間所相對的成本效益分析。最後,以健保給付者的立場來建構決策分析模型並進行成本效益分析,成本僅考量直接醫療成本,而降低FN風險所相關的結果及參數則是參考文獻及和信的資料而得。

結果 自2007年1月至2007年12月, 共有511位於和信治癌中心接受化療和lenograstim之乳癌病患納入本研究。 在這些患者中,接受高FN風險化療之病人在經過三天之G-CSF給予後(5μg/kg/day)並沒有因為接受G-CSF預防性治療而明顯降低FN之發生率(33.33%)。此外,經過資料分析後,我們發現在乳癌病患最常給予的化療處方為(1) adriamycin, paclitaxel, 及cyclophosphamide序貫療法(ATC); (2) adriamycin, paclitaxel, 及 cyclophosphamide合併療法(TAC); 和(3) cyclophosphamide, adriamycin, 及 5-fluorouracil (CAF)。在這三組接受化療及G-CSF之病患中,TAC發生FN之風險(33.33 %,初級預防)最為顯著,明顯的比ATC (6.66%,初級預防)及CAF(5.97%,次級預防)二組高出許多。而G-CSF使用的期間則分別為:CAF組-3.19天、ATC組-6.08天、TAC組-2.95天。最後,在接受不同化療處方的三組中,由於健保局的規範,大部分病人僅接受三天的G-CSF治療。經過成本效益的分析後,將三天之G-CSF療程比上七天之G-CSF療程(文獻)其增加成本效益比(ICER)明顯為高(每避免一次FN需多支出120,265元)。
討論 在乳癌病患中,接受較高FN風險之化療及G-CSF的病患,仍有很高的比率會發生FN,這可能和G-CSF使用的時間有關,我們發現接受TAC化療的病患即使給予G-CSF的預防性治療,其發生低嗜中性白血球合併發燒之機率仍遠比臨床試驗所得到之數據大約高出五倍(33.33% vs.7.5%)。此外,在健保的規範之下,臨床上使用G-CSF的天數也遠低於準則中之建議(3天vs.10~14天)。在ATC組中(6.66%),和Citron等人之研究(2%)相比,FN之發生率約高出三倍。而CAF組所發生FN之比率在早期的研究中(2~5%)及本研究(5.97%)則是相似的。在TAC組中FN發生率明顯較高,其確切原因仍是未知,但可能和使用較短期間(3天)之G-CSF有密切的關係。
結論 根據本研究的結果,在接受高風險化療處方的病患,即使接受了G-CSF的預防性治療,但仍有很高的比例會發生低嗜中性球合併發燒。這可能是由於在目前健保給付規範下,G-CSF只能使用3天的療程。此外,許多病患在接受完3天的療程後,仍需再接收另一次的3天療程以治療嗜中性白血球低下。在經過成本效益分析後,初步的結果顯示七天的G-CSF療程不只是有好的成本效益,在臨床使用上也可達到相當程度的成本節省。因此,進一步進行前瞻性研究來找出G-CSF最適當的使用天數為本研究之未來方向,並希望以本研究的結果來說服健保局以評估修正G-CSF給付規範之可能性。
英文摘要 Rationale Febrile neutropenia (FN) is a serious hematological toxicity of cancer chemotherapy and could be prevented with the prophylactic use of antibiotics and/or granulocyte colony-stimulating factor (G-CSF). G-CSF is hematopoietic growth-stimulating factor, which regulates the proliferation, differentiation and function of hematopoietic cells. G-CSF dose-dependently increases the cell number of circulating neutrophils. Early clinical trials indicated that primary prophylaxis with G-CSF reduces the duration of chemotherapy-induced neutropenia resulting in a 50% reduction in FN, infections, hospitalization, and the use of antibiotics in small cell lung cancer patients. In the study, the rate of FN was reduced from 77% to 40%, with long-term G-CSF treatment started on day 4 and continuing through day 17. Regarding to breast cancer patients, G-CSF as primary prophylaxis administered on days 4 to10 after TAC (adriamycin, paclitaxel, and cyclophosphamide) chemotherapy regimen lead to a reduction of FN risk from 27.5% to 7.5%. Based on the practice guidelines, primary prophylaxis of G-CSF treatment protocol can only be used for high risk patients with FN.

Objective G-CSF is relatively expensive medicine. Currently, at the Sun Yat-Sen Cancer Center (SYSCC), G-CSF was prescribed to patients with prior severe neutropenia or FN for only three days due to the regulation of national health insurance (NHI). The main objective of this study was to investigate the rate of FN after primary and secondary prophylaxis of G-CSF in breast cancer patients (n=511); the secondary objectives were to investigate the prescribing pattern of G-CSF at SYSCC and to evaluate the cost-effectiveness of G-CSF use under the regulation of NHI.

Study Design We performed a retrospective chart review of breast cancer patients who received chemotherapy or lenograstim at SYSCC during year 2007. The information included in this review were patients’ age, sex, tumor type, dosage of chemotherapy, the dose and duration of G-CSF, history of FN and neutropenia, and the length and cost of hospitalization due to FN. Then, we divided patients into three groups according to the risk of FN on received chemotherapy regimen, high (>20%), intermediate (10-20%), low (<10%), to find out the FN risk in different groups and also the regimen of lenograstim use. Furthermore, we focus on the FN risk and regimen of G-CSF use in patients with breast cancer who received chemotherapy regimen of TAC, ATC, and CAF. All these data will be analyzed and compared with the current literature to evaluate the efficacy and cost-effectiveness of G-CSF. Finally, a decision-analysis model was constructed from a health insurer’s perspective to evaluate the cost-effectiveness by considering direct medical costs only. The data required for the decision-analysis model were obtained from the medical literature and data from SYSCC.

Results Between January 2007 and December 2007, 511 breast patients who received both chemotherapy and lenograstim at SYSCC were enrolled in the study. Patients in the high risk group (TAC) did not have significantly reduced rate of FN (33.33%) after receiving lenograstim treatment for 3 days. Furthermore, the three common chemotherapy regimens for the treatment of breast cancer were (i) sequential treatment with doxorubicin, paclitaxel, and cyclophosphamide (ATC), (ii) combined treatment with doxorubicin, docetaxel, and cyclophosphamide (TAC), and (iii) treatment with cyclophosphamide, doxorubicin, and 5-fluorouracil (CAF). Comparison of three treatment groups, group TAC (33.33%, with primary prophylaxis G-CSF) was associated with higher FN incidence than ATC (6.66%, with primary prophylaxis G-CSF) and CAF (5.97%, with secondary prophylaxis G-CSF) groups after 3 days lenograstim treatment. And the duration of lenograstim were 3.19 days in CAF group, 6.08 days in ATC group, and 2.95 days in TAC group, respectively. Finally, most of patients in the TAC and CAF groups received only 3 days of G-CSF prophylaxis due to the regulation of national health insurance (NHI). After the cost-effectiveness analysis, the incremental cost-effectiveness ratio (ICER) in comparison of 7-day treatment regimen (according to clinical trials) versus 3-day treatment regimen was unacceptably high (N.T.120,265 per FN episode avoided).

Discussion The higher FN incidence in breast cancer patients of high risk chemotherapy regimen after G-CSF prophylaxis may be due to the short duration of G-CSF use. The risks of FN in TAC group of SYSCC and the prior study were 33.33% and 7.5%, respectively. This 5-fold higher risk in the TAC group may be due to the difference in period of G-CSF prophylaxis reducing from 7 days to 3 days in the SYSCC. In ATC group, the risk of FN in the present study (6.66%) was 3 times higher than the report of Citron et al. (2%). Earlier studies indicate the risk of developing FN in patients received CAF chemotherapy are ranging from 2 to 5 % while in the present study the risk was similar (5.97%). The reason of unexpected higher incidence of FN in TAC regimen is unclear, but is highly likely that the decreased efficacy was due to short time period (3 days) of G-CSF treatment.

Concluding Remark Based on present results, patients who received chemotherapy of high risk in FN and G-CSF for primary prophylaxis still had the rate of FN 33.33%. The result may be due to the 3-day G-CSF use under the regulation of NHI. Moreover, many patients under 3-day G-CSF regimen may further require another 3-day G-CSF regimen to support the developed neutropenia. Additionally, the cost-effectiveness model provides evidence that 7-day G-CSF regimen is not only cost-effective but also cost-saving in clinical settings. There appeared to be both clinical and economic benefits from prophylaxis with standard administration (7-day) of G-CSF. Therefore, further prospective study should be performed to find out the most appropriate duration of G-SCF use. The result could be provided as an evidence to persuade and modify the reimbursement policy of NHI.
論文目次 Acknowledgements I
Abbreviations II
Contents III
List of Tables V
List of Figures VI
List of Attachments VII
中文摘要 VIII
Abstract XI
Chapter 1 Preface 1
1.1 Rationale 1
1.2 Background 2
1.3 Purpose of the Study 2
Chapter 2 Review of Literature 5
2.1 Introduction 5
2.2 Chemotherapy-Induced Neutropenia and Febrile Neutropenia 6
2.2.1 Incidence and Risk of Febrile Neutropenia 6
2.2.2 Risk Factors of Febrile Neutropenia 10
2.3 G-CSF in Clinical Use 14
2.3.1 Clinical Efficacy of G-CSF 17
2.3.2 G-CSF Use in Chemotherapy-induced Neutropenia 19
2.3.3 Recommendations of the EORTC (2006) / ASCO (2006) / NCCN (2008) for G-CSF Use 20
2.3.4 Prophylactic G-CSF Increase the Dose Intensity and Dose-Dense Therapy of Chemotherapy 24
2.3.5 G-CSF Use in Elder Patients 24
2.4 Rational Use of G-CSF in Breast Cancer 25
2.4.1 The Importance of Taxanes in the Adjuvant Chemotherapy in Breast Cancer 25
2.5 Dose Dense Chemotherapy with G-CSF Support in the Adjuvant and Neoadjuvant Treatment for Breast Cancer Patients 29
2.5.1 Norton-Simon Hypothesis:Improving Effectiveness by Shortened Intervals 30
2.5.2 G-CSF Allows Dose-Dense Chemotherapy Regimen 32
2.6 Summary 34
Chapter 3 Experimental Design 35
3.1 Method 35
3.2 Database 36
3.3 Data Extraction: 36
3.4 Pharmacoeconomics of G-CSF 37
Chapter 4 Results 39
4.1 G-CSF Use in Most Three Common Chemotherapy Regimens of Breast Cancer 39
4.1.1 FN Risk 39
4.1.2 The Regimen of G-CSF Use in Three Chemotherapy Regimens 39
4.2 Cost-Effectiveness Analysis of G-CSF 45
Chapter 5 Discussion 47
5.1 High Risk Group have Higher FN Rate Even after Primary Prophylaxis G-CSF 47
5.2 Cost-Effectiveness of G-CSF 53
Chapter 6 Conclusion 54
Reference 55
Attachments 60

參考文獻 Aapro MS, Cameron DA, Pettengell R, Bohlius J, Crawford J, Ellis M, Kearney N, Lyman GH, Tjan-Heijnen VC, Walewski J, Weber DC and Zielinski C (2006) EORTC guidelines for the use of granulocyte-colony stimulating factor to reduce the incidence of chemotherapy-induced febrile neutropenia in adult patients with lymphomas and solid tumours. Eur J Cancer 42:2433-2453.
Avalos BR GJ, Hedvat C (1990) Human granulocyte colony-stimulating factor: biologic activities and receptor characterization on hematopoietic cells and small cell lung cancer cell lines. Blood 75:851-857.
Berghmans T, Paesmans M, Lafitte JJ, Mascaux C, Meert AP, Jacquy C, Burniat A, Steels E, Vallot F and Sculier JP (2002) Therapeutic use of granulocyte and granulocyte-macrophage colony-stimulating factors in febrile neutropenic cancer patients. A systematic review of the literature with meta-analysis. Support Care Cancer 10:181-188.
Berliner N HA, Graubert T (1995) Granulocyte colony-stimulating factor induction of normal human bone marrow progenitors results in neutrophil-specific gene expression. Blood 85:799-803.
Bethesda M (1999) National cancer Institute (NCI). http://www.cancer.gov/.
Chia S, Bryce C and Gelmon K (2005) The 2000 EBCTCG overview: a widening gap. Lancet 365:1665-1666.
Chrischilles E, Delgado DJ, Stolshek BS, Lawless G, Fridman M and Carter WB (2002) Impact of age and colony-stimulating factor use on hospital length of stay for febrile neutropenia in CHOP-treated non-Hodgkin's lymphoma. Cancer Control 9:203-211.
Citron ML, Berry DA, Cirrincione C, Hudis C, Winer EP, Gradishar WJ, Davidson NE, Martino S, Livingston R, Ingle JN, Perez EA, Carpenter J, Hurd D, Holland JF, Smith BL, Sartor CI, Leung EH, Abrams J, Schilsky RL, Muss HB and Norton L (2003) Randomized trial of dose-dense versus conventionally scheduled and sequential versus concurrent combination chemotherapy as postoperative adjuvant treatment of node-positive primary breast cancer: first report of Intergroup Trial C9741/Cancer and Leukemia Group B Trial 9741. J Clin Oncol 21:1431-1439.
Crawford J (2008) Myeloid Growth Factors. NCCN;Practice Guidelinesin Oncology v.1.
Crawford J, Dale DC and Lyman GH (2004) Chemotherapy-induced neutropenia: risks, consequences, and new directions for its management. Cancer 100:228-237.
Crawford J, Ozer H, Stoller R, Johnson D, Lyman G, Tabbara I, Kris M, Grous J, Picozzi V, Rausch G and et al. (1991) Reduction by granulocyte colony-stimulating factor of fever and neutropenia induced by chemotherapy in patients with small-cell lung cancer. N Engl J Med 325:164-170.
Dale DC (2002) Colony-stimulating factors for the management of neutropenia in cancer patients. Drugs 62 Suppl 1:1-15.
Dale DC BM, Davis MW (1993) A randomized controlled phase III trial of recombinant human granulocyte colony-stimulating factor (filgrastim) for treatment of severe chronic neutropenia. Blood 81:2496-2502.
Demetri GD GJ (1991) Granulocyte colony-stimulating factor and its receptor. Blood 78:2791-2808.
EBCTCG (1998) Polychemotherapy for early breast cancer: an overview of the randomised trials. . Lancet 352:930-942.
Garcia-Carbonero R, Mayordomo JI, Tornamira MV, Lopez-Brea M, Rueda A, Guillem V, Arcediano A, Yubero A, Ribera F, Gomez C, Tres A, Perez-Gracia JL, Lumbreras C, Hornedo J, Cortes-Funes H and Paz-Ares L (2001) Granulocyte colony-stimulating factor in the treatment of high-risk febrile neutropenia: a multicenter randomized trial. J Natl Cancer Inst 93:31-38.
GJ L (1997) CSF-deficient mice - what have they taught us? Ciba Found Symp 204:60-74.
Glaspy JA, Bleecker G, Crawford J, Stoller R and Strauss M (1993) The impact of therapy with filgrastim (recombinant granulocyte colony-stimulating factor) on the health care costs associated with cancer chemotherapy. Eur J Cancer 29A Suppl 7:S23-30.
Haim N, Shulman K, Goldberg H and Tsalic M (2005) The safety of full-dose chemotherapy with secondary prophylactic granulocyte colony stimulating factor (G-CSF) following a prior cycle with febrile neutropenia. Med Oncol 22:229-232.
Hartmann LC, Tschetter LK, Habermann TM, Ebbert LP, Johnson PS, Mailliard JA, Levitt R, Suman VJ, Witzig TE, Wieand HS, Miller LL and Moertel CG (1997) Granulocyte colony-stimulating factor in severe chemotherapy-induced afebrile neutropenia. N Engl J Med 336:1776-1780.
Henderson IC, Berry DA, Demetri GD, Cirrincione CT, Goldstein LJ, Martino S, Ingle JN, Cooper MR, Hayes DF, Tkaczuk KH, Fleming G, Holland JF, Duggan DB, Carpenter JT, Frei E, 3rd, Schilsky RL, Wood WC, Muss HB and Norton L (2003) Improved outcomes from adding sequential Paclitaxel but not from escalating Doxorubicin dose in an adjuvant chemotherapy regimen for patients with node-positive primary breast cancer. J Clin Oncol 21:976-983.
Hsieh MM, Everhart JE, Byrd-Holt DD, Tisdale JF and Rodgers GP (2007) Prevalence of neutropenia in the U.S. population: age, sex, smoking status, and ethnic differences. Ann Intern Med 146:486-492.
Klastersky J and Paesmans M (2007) Risk-adapted strategy for the management of febrile neutropenia in cancer patients. Support Care Cancer 15:477-482.
Kuderer NM, Dale DC, Crawford J, Cosler LE and Lyman GH (2006) Mortality, morbidity, and cost associated with febrile neutropenia in adult cancer patients. Cancer 106:2258-2266.
Kuderer NM, Dale DC, Crawford J and Lyman GH (2007) Impact of primary prophylaxis with granulocyte colony-stimulating factor on febrile neutropenia and mortality in adult cancer patients receiving chemotherapy: a systematic review. J Clin Oncol 25:3158-3167.
Kummel S KJ, Kohls A. (2006) Randomized trial: survival benefit and safety of adjuvant dose-dense chemotherapy for node-positive breast cancer. British Journal of Cancer 94:1237-1244.
Lapidot T PI (2002) Current understanding of stem cell mobilization: the roles of chemo- kines, proteolytic enzymes, adhesion molecules, cytokines, and stromal cells. Exp Hematol 30:973-981.
Lewis JP (2007) An interpretation of the EBCTCG data. Oncologist 12:505-509.
Link H, Bohme A, Cornely OA, Hoffken K, Kellner O, Kern WV, Mahlberg R, Maschmeyer G, Nowrousian MR, Ostermann H, Ruhnke M, Sezer O, Schiel X, Wilhelm M and Auner HW (2003) Antimicrobial therapy of unexplained fever in neutropenic patients--guidelines of the Infectious Diseases Working Party (AGIHO) of the German Society of Hematology and Oncology (DGHO), Study Group Interventional Therapy of Unexplained Fever, Arbeitsgemeinschaft Supportivmassnahmen in der Onkologie (ASO) of the Deutsche Krebsgesellschaft (DKG-German Cancer Society). Ann Hematol 82 Suppl 2:S105-117.
Liu F WH, Wesselschmidt R, Kornaga T, Link DC (1996) Impaired production and increased apoptosis of neutrophils in granulocyte colony-stimulating factor receptor-deficient mice. Immunity 5:491-501.
Lowenberg B vPW, Theobald M (2003) Effect of Priming with Granulocyte Colony- Stimulating Factor on the Outcome of Chemotherapy for Acute Myeloid Leukemia. N Engl J Med 349:743-752.
Lyman GH, Lyman CH and Agboola O (2005) Risk models for predicting chemotherapy-induced neutropenia. Oncologist 10:427-437.
Mamounas EP, Bryant J, Lembersky B, Fehrenbacher L, Sedlacek SM, Fisher B, Wickerham DL, Yothers G, Soran A and Wolmark N (2005) Paclitaxel after doxorubicin plus cyclophosphamide as adjuvant chemotherapy for node-positive breast cancer: results from NSABP B-28. J Clin Oncol 23:3686-3696.
Martin M, Lluch A, Segui MA, Ruiz A, Ramos M, Adrover E, Rodriguez-Lescure A, Grosse R, Calvo L, Fernandez-Chacon C, Roset M, Anton A, Isla D, del Prado PM, Iglesias L, Zaluski J, Arcusa A, Lopez-Vega JM, Munoz M and Mel JR (2006) Toxicity and health-related quality of life in breast cancer patients receiving adjuvant docetaxel, doxorubicin, cyclophosphamide (TAC) or 5-fluorouracil, doxorubicin and cyclophosphamide (FAC): impact of adding primary prophylactic granulocyte-colony stimulating factor to the TAC regimen. Ann Oncol 17:1205-1212.
Martin M PT, Mackey J. (2005) Adjuvant docetaxel for node-positive breast cancer. N Engl J Med 352:2302-2313.
Molineux G PZ, Hampson IN, Lord BI, Dexter TM (1990) Transplantation potential of peri- pheral blood stem cells induced by granulocyte colony-stimulating factor. Blood 76:2153-2158.
Morishita K TM, Asano S, Kaziro Y, Nagata S (1987) Chromosomal gene structure of human myeloperoxidase and regulation of its expression by granulocyte colony-stimulating factor. J Biol Chem 262:15208-15213.
NM K (2005) Meta-analysis of prophylactic granulocyte colony-stimulating factor (G-CSF) in cancer patients receiving chemotherapy. ASCO:Abstract #8117.
Norton L (1997) Evolving concepts in the systemic drug therapy of breast cancer. Semin Oncol 24:S10-13-S10-10.
Osby E, Hagberg H, Kvaloy S, Teerenhovi L, Anderson H, Cavallin-Stahl E, Holte H, Myhre J, Pertovaara H and Bjorkholm M (2003) CHOP is superior to CNOP in elderly patients with aggressive lymphoma while outcome is unaffected by filgrastim treatment: results of a Nordic Lymphoma Group randomized trial. Blood 101:3840-3848.
Pfreundschuh M, Trumper L, Kloess M, Schmits R, Feller AC, Rube C, Rudolph C, Reiser M, Hossfeld DK, Eimermacher H, Hasenclever D, Schmitz N and Loeffler M (2004) Two-weekly or 3-weekly CHOP chemotherapy with or without etoposide for the treatment of elderly patients with aggressive lymphomas: results of the NHL-B2 trial of the DSHNHL. Blood 104:634-641.
Rigacci L, Nassi L, Alterini R, Carrai V, Longo G, Bernardi F, Martini V and Bosi A (2006) Dose-dense CHOP plus rituximab (R-CHOP14) for the treatment of elderly patients with high-risk diffuse large B cell lymphoma: a pilot study. Acta Haematol 115:22-27.
Rivera E, Erder MH, Moore TD, Shiftan TL, Knight CA, Fridman M, Brannan C, Danel-Moore L and Hortobagyi GN (2003a) Targeted filgrastim support in patients with early-stage breast carcinoma: toward the implementation of a risk model. Cancer 98:222-228.
Rivera E, Haim Erder M, Fridman M, Frye D and Hortobagyi GN (2003b) First-cycle absolute neutrophil count can be used to improve chemotherapy-dose delivery and reduce the risk of febrile neutropenia in patients receiving adjuvant therapy: a validation study. Breast Cancer Res 5:R114-120.
Saintigny P, Assouad S, Gligorov J, Selle F, Roche H, Breau JL, Morere JF and Lotz JP (2004) [Dose density and dose intensity in the treatment of breast cancer]. Bull Cancer 91 Suppl 4:S244-253.
Sato N AS, Koeffler HP, Yoshida S, Takaku F, Takatani O (1988) Identification of neutrophil alkaline phosphatase-inducing factor in cystic fluid of a human squamous cell carcinoma as granulocyte colony-stimulating factor. J Cell Physiol 137:272-276.
Semerad CL LF, Gregory AD, Stumpf K, Link DC (2002) G-CSF Is an Essential Regulator of Neutrophil Trafficking from the Bone Marrow to the Blood. Immunity 17:413.
Shayne M, Culakova E, Poniewierski MS, Wolff D, Dale DC, Crawford J and Lyman GH (2007) Dose intensity and hematologic toxicity in older cancer patients receiving systemic chemotherapy. Cancer 110:1611-1620.
Silber JH, Fridman M, DiPaola RS, Erder MH, Pauly MV and Fox KR (1998) First-cycle blood counts and subsequent neutropenia, dose reduction, or delay in early-stage breast cancer therapy. J Clin Oncol 16:2392-2400.
Smith TJ, Khatcheressian J, Lyman GH, Ozer H, Armitage JO, Balducci L, Bennett CL, Cantor SB, Crawford J, Cross SJ, Demetri G, Desch CE, Pizzo PA, Schiffer CA, Schwartzberg L, Somerfield MR, Somlo G, Wade JC, Wade JL, Winn RJ, Wozniak AJ and Wolff AC (2006) 2006 update of recommendations for the use of white blood cell growth factors: an evidence-based clinical practice guideline. J Clin Oncol 24:3187-3205.
Thatcher N, Girling DJ, Hopwood P, Sambrook RJ, Qian W and Stephens RJ (2000) Improving survival without reducing quality of life in small-cell lung cancer patients by increasing the dose-intensity of chemotherapy with granulocyte colony-stimulating factor support: results of a British Medical Research Council Multicenter Randomized Trial. Medical Research Council Lung Cancer Working Party. J Clin Oncol 18:395-404.
Thomas J LF, Link DC (2002) Mechanisms of mobilization of hematopoietic progenitors with granulocyte colony-stimulating factor. Curr Opin Hematol 9:183-189.
Trillet (1993) Pathol Biol (Paris):Jan;41(41):46.
Wells JA and de Vos AM (1996) Hematopoietic receptor complexes. Annu Rev Biochem 65:609-634.
Welte K BL (1997) Severe chronic neutropenia: pathophysiology and therapy. Semin Hematol 34:267-278.
Welte K ZC, Reiter A (1990) Differential effects of granulocyte-macrophage colony- stimulating factor and granulocyte colony-stimulating factor in children with severe congenital neutropenia. Blood 75:1056-1063.
Woll PJ, Hodgetts J, Lomax L, Bildet F, Cour-Chabernaud V and Thatcher N (1995) Can cytotoxic dose-intensity be increased by using granulocyte colony-stimulating factor? A randomized controlled trial of lenograstim in small-cell lung cancer. J Clin Oncol 13:652-659.
Zhan Y LG, Grail D, Dunn AR, Cheers C (1998) Essential roles for granulocyte-macrophage colony-stimulating factor (GM-CSF) and G-CSF in the sustained hematopoietic response of Listeria monocytogenes-infected mice. Blood 91:863-869.

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系統識別號 U0007-0707200915583600
論文名稱(中文) 分析抗焦慮劑/安眠劑之使用的影響因子在重度憂鬱症及廣泛性焦慮症病人和一般大眾的處方形態
論文名稱(英文) Analysis of Factors Associated with the Use of Anxiolyitcs/Hypnotics among MDD and GAD Patients and the Prescription Pattern among General Population
校院名稱 臺北醫學大學
系所名稱(中) 藥學研究所
系所名稱(英) Graduate Institute of Pharmacy
學年度 97
學期 2
出版年 98
研究生(中文) 江卓倫
學號 M301096019
學位類別 碩士
語文別 英文
口試日期 2009-06-15
論文頁數 76頁
口試委員 指導教授-沈武典
共同指導教授-闕壯卿
委員-李信謙
委員-陳楚杰
委員-吳姿樺
關鍵字(中) 苯二氮平
安眠鎮靜劑
處方形態
全民健康保險資料庫
萬芳醫學中心資料庫
苯二氮平之使用治療指引
關鍵字(英) benzodiazepine
hypnotic
anxiolytic
prescription pattern
National Health Insurance Research Database (NHIRD)
TMU-Wan Fang Medical Center database (T-WFMCD)
benzodiazepine guidelines
學科別分類
中文摘要 研究背景: 苯二氮平(benzodiazepine)和苯二氮平接受器致效劑(benzodiazepine receptor agonist,俗稱Z-drug)這類的抗焦慮劑和鎮靜安眠劑通常被用來在重度憂鬱症(major depressive disorder)及廣泛性焦慮症 (generalized anxiety disorder) 的病人中,緩解病人焦慮以及失眠的情形。使用鎮靜安眠劑所要考量到的是,它可能會有耐藥性(tolerance)、成癮(dependence)、車禍或是易造成老年人跌倒的危險;所以治療指引建議這類僅使用在病人處於急性期的狀態。過去研究指出,在某些的族群中(特別是老年人),哪些因子會影響這類藥物的使用。然而在重度憂鬱症及廣泛性焦慮症的病人族群中,這樣的研究則是很少。台灣在2005年發行BZD的使用治療指引,但沒有研究指出醫師是否有遵照此指引來開立BZD的處方。

研究目的: 本研究主要的目的是要去觀察抗焦慮劑/安眠劑的使用,在重度憂鬱症及廣泛性焦慮症的病人中;以及利用2004及2006年台灣全民健康保險資料庫來比較抗焦慮劑/安眠劑的使用狀況。

研究方法: 本研究的資料來源為全民健康保險資料庫(2004-2006年),以及臺北醫學大學•萬芳醫學中心的資料庫(2004年-2009年1月)。本研究分成兩個部份。第一個部份,研究的對象為有被診斷為重度憂鬱症(major depressive disorder),且或者是廣泛性焦慮症(generalized anxiety disorder)的病人;病人如果其它疾病像是癲癇(seizure),或者是中樞系統的疾病,則會被排除,因為這些疾病可能會影響病人使用鎮靜安眠劑的時間。而萬芳醫學中心組的部份,病人如能經由醫生指示完成抗憂鬱劑的治療療程,則納入。本研究使用單因子迴歸分析 (univariate regression model) 來選擇與抗焦慮劑/安眠藥使用有顯著相關之因子,然後使用多因子逐步分析迴歸模式(backward stepwise multivariate regression model)來找出對抗焦慮劑/安眠藥使用更具有影響力的相關因子。在本研究的第二部份,以2004及2006年全民健康保險資料庫為資料來源,每一年資料庫提供一百萬人的資料;將有拿慢性處方籤之病人資料收集,用來比較在治療指引發行前一年及後一年,藥品處方的情況有何不同。本研究使用卡方檢定(chi-square test) 來分析其統計上的差異。

研究結果: 在本研究的第一個部份,在2004 到2006年期間的全民健保資料庫中,有17,266位病人被診斷為重度憂鬱症或廣泛性焦慮症;其中有8,902位病人是重度憂鬱症,有9,334位病人是廣泛性焦慮症,而有1,010位病人同時被診斷有這種疾病。將這些病人符合本研究之納入、排除條件之後,共399位病人被納入本研究的健保資料庫組,而有63位病人被納入至萬芳醫學中心組。在兩組資料庫中,抗憂鬱劑的開立天數及抗憂鬱劑的使用個數是兩個具有顯著相關的因子。年齡及大於四個月的抗焦慮劑/安眠劑的使用在抗憂鬱劑的治療前,則有顯著相關在健保資料庫組。在本研究的第二部份中,在每年一百萬人的全民健康保險資料庫中,在2004年有11,039位病人有拿BZD的慢性處方籤,而在2006年則顯著上升至16,193位病人。而有拿開立BZD超過30天的慢性處方籤,在2004年有6,505位病人,在2006年則顯著上升至8,790位病人。另一方面,有拿開立BZD 30天以下之病人,在2004年有4,534位病人,在2006年則顯著上升至7,401位病人。

研究結論: 本研究的結果顯示,抗憂鬱劑的開立天數及抗憂鬱劑的使用個數對抗焦慮劑/安眠劑的使用有顯著的相關性。而年齡及大於四個月的抗焦慮劑/安眠劑的使用在抗憂鬱劑的治療前,有顯著的相關性在健保資料庫組中。在治療指引發行之後,2006年的BZD處方量比較在指引發行前的2004年,顯著的上升。明顯地,在台灣醫師開立BZD的處方行為上,並沒有因為2005年指引的發行而有所改變。
英文摘要 Background: Benzodiazepines (BZDs) or benzodiazepine receptor agonists, also called Z-drugs (e.g. zolpidem, zopiclone or zaloplon), have anxiolytic and hypnotic functions to relieve symptoms among major depressive disorder (MDD) and generalized anxiety disorder (GAD) patients. The concerns of using anxiolytic and hypnotic agents are tolerance, dependence, accident proneness, and increased risk of falls in elderly. Thus, these agents should be used only in the acute phase, indicated by treatment guidelines. Factors associated with the use of these agents have been studied within whole population, especially in the elderly. However, the factors have not been studied in MDD and GAD patients. Taiwan’s government authority implemented the guideline in 2005. Whether physicians would have followed this guideline to prescribe BZDs, is still unknown.
Objective: The goal was to investigate the use of anxiolytic/hypnotic agents in the MDD and GAD patients and to compare the prescription pattern of anxiolytic/hypnotic agents in National Health Insurance Research Database (NHIRD) 2004 and 2006 in Taiwan.
Methods: The data of this study were obtained from NHIRD (for 2004 to 2006; 1,000,000 people per year) and TMU-Wan Fang Medical Center database (T-WFMCD) during 2004 to 2009. In the first part of this study, patients who were diagnosed of either MDD or GAD at clinics were selected, and those who had other medical conditions (e.g. seizures or other CNS disorders) that affect the duration of hypnotic-use were excluded. Patients who had finished their antidepressant treatment with doctor’s guide were included in T-WFMCD group. Univariate regression models were performed to select the significant correlated factors with the use of anxiolytic/hypnotic agents. Then backward stepwise multivariate regression models were performed to analyze the factors with the more powerful effect on the use of anxiolytic/hypnotic agents among these significant correlated factors. In the second part of this study, the data were based on the 1,000,000 people in 2004 and 2006 NHIRD. The numbers of patients who received the BZD prescriptions in chronic use were collected for comparison of the prescription pattern in pre- and post-guideline year. Chi-square test was used for testing the statistic difference.
Results: In the first part of this study, 17,226 patients were diagnosed of MDD and/or GAD during 2004 to 2006. The sum of 8,902 and 9,334 patients were diagnosed as MDD and GAD, respectively in these three years. A total of 1,010 patients were comorbid with both MDD and GAD. After fulfilling the inclusion and exclusion criteria, 399 patients were included into NHIRD group. 63 patients were included into T-WFMCD group. The antidepressant-prescribing days, and the number of antidepressants used were found to be significantly correlated in both databases. Age and the four month or more of the use of anxiolytic/hypnotic agents before antidepressant therapy were found to be significantly correlated in NHIRD group., Based on the 1,000,000 patient population in the second part of this study, the overall numbers of patients who received the BZD prescriptions in chronic use were found to be increased significantly from 11,039 to 16,193 in 2004 to 2006. And the numbers of patients who received more than 30 days of BZDs were increased significantly from 6,505 to 8,790. While the numbers of patients with less than 30 days of BZDs were found to be increased significantly from 4,534 to 7,401.
Conclusion: The results of present study suggested that the antidepressant-prescribing days, and the numbers of antidepressants used are significant associated with the use of anxiolytic/hypnotic agents. And age and the four month or more of the use of anxiolytic/hypnotic agents have significant association in NHIRD group. After the publication of the guideline, the BZDs prescriptions in 2006 have been found to be significantly increased over the pre-guideline year of 2004. Apparently the prescription pattern of BZDs by Taiwanese physicians was not changed by the publication of the 2005 guideline of the management and use of BZDs.
論文目次 博碩士論文授權書 i
臺北醫學大學碩士班研究生論文口試委員審定書 ii
Abstract iii
Summary Chinese vi
Acknowledgements x
Contents xii
List of Tables xiv
List of Figures xvi
Abbreviation List xvii
Chapter 1 Preface 1
1.1. Rationale 1
1.2. Research Goal 3
Chapter 2 Introduction 4
2.1. Subject 4
2.1.1. Major Depressive Disorder 4
2.1.2. Generalized Anxiety Disorder 5
2.2. Epidemiology of Major Depressive Disorder and Generalized Anxiety Disorder 5
2.3. Pharmacotherapy for Major Depressive Disorder and Generalized Anxiety Disorder 6
2.4. Practice guidelines on using the benzodiazepines 8
2.5. Clinical Issues 14
2.6. New Study Direction 16
Chapter 3 Experimental Design 18
3.1. Study Subjects – The First Part of this Study 18
3.1.1. Inclusion criteria in NHIRD 18
3.1.2. Inclusion criteria at Wan Fang Medical Center 18
3.1.3. Exclusion criteria with patients in both databases 19
3.2. Study Subjects – the Second Part of this Study 19
3.3. Databank 20
3.4. Data retrieval – the First Part of this Study 21
3.4.1. Data retrieval from NHIRD 21
3.4.2. Data retrieval in the database of Wan Fang Medical Center 25
3.5. Data retrieval – the Second Part of this Study 25
3.6. The Definitions of the Variables in the First Part of this Study 26
3.6.1. Dependent Variables 26
3.6.2. Independent Variables 26
3.7. Statistic Methods 32
Chapter 4 Results 33
4.1. Result in the First Part of this Study 33
4.1.1. National Insurance Health Research Database 33
4.1.2. Post-hoc Analysis in the NIHRD 41
4.1.3. The Database of Wan Fang Medical Center 43
4.1.4. Post-hoc Analysis in the Database of Wan Fang Medical Center 48
4.2. The Results in the Second Part of this Study 53
Chapter 5 Discussion 58
5.1. Original Mistake in Experimental Design of this Study 58
5.2. Discussion of the Factors Associated with the Use of Anxiolytic/hypnotic Agents (the First Part of this Study) 59
5.3. Discussion of the Management and Use of hypnotics in Taiwan (The Second Part of this Study) 64
Chapter 6 Concluding Remarks 67
References 68
Appendix 74

參考文獻 Alvarenga JM, Loyola Filho AI, Firmo JO, Lima-Costa MF and Uchoa E (2008) Prevalence and sociodemographic characteristics associated with benzodiazepines use among community dwelling older adults: the Bambui Health and Aging Study (BHAS). Rev Bras Psiquiatr 30:7-11.
APA (2000a) American Psychiatric Association: Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision. Washington, DC, American Psychiatric Association, 2000.
APA (2000b) Practice guideline for the treatment of patients with major depressive disorder (revision). American Psychiatric Association, in Am J Psychiatry pp 1-45.
Ashton H (1994) Guidelines for the rational use of benzodiazepines. When and what to use. Drugs 48:25-40.
Beck CA, Williams JV, Wang JL, Kassam A, El-Guebaly N, Currie SR, Maxwell CJ and Patten SB (2005) Psychotropic medication use in Canada. Can J Psychiatry 50:605-613.
Benitez CI, Smith K, Vasile RG, Rende R, Edelen MO and Keller MB (2008) Use of benzodiazepines and selective serotonin reuptake inhibitors in middle-aged and older adults with anxiety disorders: a longitudinal and prospective study. Am J Geriatr Psychiatry 16:5-13.
Berman RM, Marcus RN, Swanink R, McQuade RD, Carson WH, Corey-Lisle PK and Khan A (2007) The efficacy and safety of aripiprazole as adjunctive therapy in major depressive disorder: a multicenter, randomized, double-blind, placebo-controlled study. J Clin Psychiatry 68:843-853.
Bose A, Korotzer A, Gommoll C and Li D (2008) Randomized placebo-controlled trial of escitalopram and venlafaxine XR in the treatment of generalized anxiety disorder. Depress Anxiety 25:854-861.
Chen TJ, Chou LF and Hwang SJ (2006) Patterns of ambulatory care utilization in Taiwan. BMC Health Serv Res 6:54.
Cheng JS, Huang WF, Lin KM and Shih YT (2008) Characteristics associated with benzodiazepine usage in elderly outpatients in Taiwan. Int J Geriatr Psychiatry 23:618-624.
Chien IC, Kuo CC, Bih SH, Chou YJ, Lin CH, Lee CH and Chou P (2007) The prevalence and incidence of treated major depressive disorder among National Health Insurance enrollees in Taiwan, 1996 to 2003. Can J Psychiatry 52:28-36.
Cimolai N (2007) Zopiclone: is it a pharmacologic agent for abuse? Can Fam Physician 53:2124-2129.
Davidson J, Allgulander C, Pollack MH, Hartford J, Erickson JS, Russell JM, Perahia D, Wohlreich MM, Carlson J and Raskin J (2008) Efficacy and tolerability of duloxetine in elderly patients with generalized anxiety disorder: a pooled analysis of four randomized, double-blind, placebo-controlled studies. Hum Psychopharmacol 23:519-526.
de las Cuevas C, Sanz E and de la Fuente J (2003) Benzodiazepines: more "behavioural" addiction than dependence. Psychopharmacology (Berl) 167:297-303.
Demyttenaere K, Bonnewyn A, Bruffaerts R, De Girolamo G, Gasquet I, Kovess V, Haro JM and Alonso J (2008) Clinical factors influencing the prescription of antidepressants and benzodiazepines: results from the European study of the epidemiology of mental disorders (ESEMeD). J Affect Disord 110:84-93.
Feltner DE, Crockatt JG, Dubovsky SJ, Cohn CK, Shrivastava RK, Targum SD, Liu-Dumaw M, Carter CM and Pande AC (2003) A randomized, double-blind, placebo-controlled, fixed-dose, multicenter study of pregabalin in patients with generalized anxiety disorder. J Clin Psychopharmacol 23:240-249.
Fortin D, Preville M, Ducharme C, Hebert R, Trottier L, Gregoire JP, Allard J and Berard A (2007) Factors associated with long-term benzodiazepine use among elderly women and men in Quebec. J Women Aging 19:37-52.
Furukawa TA, Streiner DL and Young LT (2001) Is antidepressant-benzodiazepine combination therapy clinically more useful? A meta-analytic study. J Affect Disord 65:173-177.
Furukawa TA, Streiner DL and Young LT (2002) Antidepressant and benzodiazepine for major depression. Cochrane Database Syst Rev:CD001026.
Garakani A, Martinez JM, Marcus S, Weaver J, Rickels K, Fava M and Hirschowitz J (2008) A randomized, double-blind, and placebo-controlled trial of quetiapine augmentation of fluoxetine in major depressive disorder. Int Clin Psychopharmacol 23:269-275.
Haw C and Stubbs J (2007) Benzodiazepines--a necessary evil? A survey of prescribing at a specialist UK psychiatric hospital. J Psychopharmacol 21:645-649.
Hermos JA, Young MM, Lawler EV, Stedman MR, Gagnon DR and Fiore LD (2005) Characterizations of long-term anxiolytic benzodiazepine prescriptions in veteran patients. J Clin Psychopharmacol 25:600-604.
Holbrook A, Crowther R, Lotter A and Endeshaw Y (2001) The role of benzodiazepines in the treatment of insomnia: meta-analysis of benzodiazepine use in the treatment of insomnia. J Am Geriatr Soc 49:824-826.
Huang WF and Lai IC (2005) Patterns of sleep-related medications prescribed to elderly outpatients with insomnia in Taiwan. Drugs Aging 22:957-965.
Huang WF and Lai IC (2006) Potentially inappropriate prescribing for insomnia in elderly outpatients in Taiwan. Int J Clin Pharmacol Ther 44:335-342.
Jefferson JW, Rush AJ, Nelson JC, VanMeter SA, Krishen A, Hampton KD, Wightman DS and Modell JG (2006) Extended-release bupropion for patients with major depressive disorder presenting with symptoms of reduced energy, pleasure, and interest: findings from a randomized, double-blind, placebo-controlled study. J Clin Psychiatry 67:865-873.
Katon W, Cantrell CR, Sokol MC, Chiao E and Gdovin JM (2005) Impact of antidepressant drug adherence on comorbid medication use and resource utilization. Arch Intern Med 165:2497-2503.
Kornstein SG, Dunner DL, Meyers AL, Whitmyer VG, Mallinckrodt CH, Wohlreich MM, Detke MJ, Hollandbeck MS and Greist JH (2008) A randomized, double-blind study of increasing or maintaining duloxetine dose in patients without remission of major depressive disorder after initial duloxetine therapy. J Clin Psychiatry 69:1383-1392.
Lader M (1984) Short-term versus long-term benzodiazepine therapy. Curr Med Res Opin 8 Suppl 4:120-126.
Lader M and Russell J (1993) Guidelines for the prevention and treatment of benzodiazepine dependence: summary of a report from the Mental Health Foundation. Addiction 88:1707-1708.
Lee P, Shu L, Xu X, Wang CY, Lee MS, Liu CY, Hong JP, Ruschel S, Raskin J, Colman SA and Harrison GA (2007) Once-daily duloxetine 60 mg in the treatment of major depressive disorder: multicenter, double-blind, randomized, paroxetine-controlled, non-inferiority trial in China, Korea, Taiwan and Brazil. Psychiatry Clin Neurosci 61:295-307.
Luijendijk HJ, Tiemeier H, Hofman A, Heeringa J and Stricker BH (2008) Determinants of chronic benzodiazepine use in the elderly: a longitudinal study. Br J Clin Pharmacol 65:593-599.
Mahmoud RA, Pandina GJ, Turkoz I, Kosik-Gonzalez C, Canuso CM, Kujawa MJ and Gharabawi-Garibaldi GM (2007) Risperidone for treatment-refractory major depressive disorder: a randomized trial. Ann Intern Med 147:593-602.
Marcus RN, McQuade RD, Carson WH, Hennicken D, Fava M, Simon JS, Trivedi MH, Thase ME and Berman RM (2008) The efficacy and safety of aripiprazole as adjunctive therapy in major depressive disorder: a second multicenter, randomized, double-blind, placebo-controlled study. J Clin Psychopharmacol 28:156-165.
Moore N, Verdoux H and Fantino B (2005) Prospective, multicentre, randomized, double-blind study of the efficacy of escitalopram versus citalopram in outpatient treatment of major depressive disorder. Int Clin Psychopharmacol 20:131-137.
Morin CM, Belanger L and Bernier F (2004) Correlates of benzodiazepine use in individuals with insomnia. Sleep Med 5:457-462.
Munizza C, Olivieri L, Di Loreto G and Dionisio P (2006) A comparative, randomized, double-blind study of trazodone prolonged-release and sertraline in the treatment of major depressive disorder. Curr Med Res Opin 22:1703-1713.
Murphy JM, Laird NM, Monson RR, Sobol AM and Leighton AH (2000) A 40-year perspective on the prevalence of depression: the Stirling County Study. Arch Gen Psychiatry 57:209-215.
NICE (2004) National Institute for Clinical Excellence. Guidance on the use of zaleplon, zolpidem and zopiclone for the short-term management of insomnia
Nimatoudis I, Zissis NP, Kogeorgos J, Theodoropoulou S, Vidalis A and Kaprinis G (2004) Remission rates with venlafaxine extended release in Greek outpatients with generalized anxiety disorder. A double-blind, randomized, placebo controlled study. Int Clin Psychopharmacol 19:331-336.
Pandina GJ, Canuso CM, Turkoz I, Kujawa M and Mahmoud RA (2007) Adjunctive risperidone in the treatment of generalized anxiety disorder: a double-blind, prospective, placebo-controlled, randomized trial. Psychopharmacol Bull 40:41-57.
Pollack MH, Simon NM, Zalta AK, Worthington JJ, Hoge EA, Mick E, Kinrys G and Oppenheimer J (2006) Olanzapine augmentation of fluoxetine for refractory generalized anxiety disorder: a placebo controlled study. Biol Psychiatry 59:211-215.
Reeves H, Batra S, May RS, Zhang R, Dahl DC and Li X (2008) Efficacy of risperidone augmentation to antidepressants in the management of suicidality in major depressive disorder: a randomized, double-blind, placebo-controlled pilot study. J Clin Psychiatry 69:1228-1336.
Rudolph RL, Fabre LF, Feighner JP, Rickels K, Entsuah R and Derivan AT (1998) A randomized, placebo-controlled, dose-response trial of venlafaxine hydrochloride in the treatment of major depression. J Clin Psychiatry 59:116-122.
Ruhe HG, Booij J, v Weert HC, Reitsma JB, Fransen EJ, Michel MC and Schene AH (2009) Evidence why paroxetine dose escalation is not effective in major depressive disorder: a randomized controlled trial with assessment of serotonin transporter occupancy. Neuropsychopharmacology 34:999-1010.
Salzman C (1991) The APA Task Force report on benzodiazepine dependence, toxicity, and abuse. Am J Psychiatry 148:151-152.
Salzman C (1998) Addiction to benzodiazepines. Psychiatr Q 69:251-261.
Shelton RC, Haman KL, Rapaport MH, Kiev A, Smith WT, Hirschfeld RM, Lydiard RB, Zajecka JM and Dunner DL (2006) A randomized, double-blind, active-control study of sertraline versus venlafaxine XR in major depressive disorder. J Clin Psychiatry 67:1674-1681.
Shen WW, Chang C, Hsieh WC, Yeh CJ, Chiu FY and Chuang YC (2002) The flunitrazepam abuse prevention program at a general hospital in Taiwan: a descriptive study. Psychiatry Clin Neurosci 56:425-430.
Stein DJ, Ahokas AA and de Bodinat C (2008) Efficacy of agomelatine in generalized anxiety disorder: a randomized, double-blind, placebo-controlled study. J Clin Psychopharmacol 28:561-566.
Steiner JF and Prochazka AV (1997) The assessment of refill compliance using pharmacy records: methods, validity, and applications. J Clin Epidemiol 50:105-116.
Taiwan-DOH (2005) Guidelines for Using Benzodiazepines as Anxiolytic or Sedative Drugs, in.
Taiwan: Department of Health EY (2005) Guidelines for Using Benzodiazepines as Anxiolytic or Sedative Drugs, in.
Taiwan: Department of Health EY (2007) Guidelines for Using Benzodiazepines as Anxiolytic or Sedative Drugs, in.
Valenstein M, Taylor KK, Austin K, Kales HC, McCarthy JF and Blow FC (2004) Benzodiazepine use among depressed patients treated in mental health settings. Am J Psychiatry 161:654-661.
van Rijswijk E, Zandstra SM, van de Lisdonk EH, Zitman FG and van Weel C (2005) Appropriateness of benzodiazepine prescribing in general practice. Int J Clin Pharmacol Ther 43:411-412.
von Knorring AL, Olsson GI, Thomsen PH, Lemming OM and Hulten A (2006) A randomized, double-blind, placebo-controlled study of citalopram in adolescents with major depressive disorder. J Clin Psychopharmacol 26:311-315.
Wade A, Crawford GM, Angus M, Wilson R and Hamilton L (2003) A randomized, double-blind, 24-week study comparing the efficacy and tolerability of mirtazapine and paroxetine in depressed patients in primary care. Int Clin Psychopharmacol 18:133-141.
WHO (2004) Global Burden of Disease 2004; Part III: Disease incidence, prevalence and disability, in pp 27-37, World Health Orgranization.
Zhan C, Sangl J, Bierman AS, Miller MR, Friedman B, Wickizer SW and Meyer GS (2001) Potentially inappropriate medication use in the community-dwelling elderly: findings from the 1996 Medical Expenditure Panel Survey. JAMA 286:2823-2829.
Zitman FG and Couvee JE (2001) Chronic benzodiazepine use in general practice patients with depression: an evaluation of controlled treatment and taper-off: report on behalf of the Dutch Chronic Benzodiazepine Working Group. Br J Psychiatry 178:317-324.

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系統識別號 U0007-0807200910204400
論文名稱(中文) 探討懷孕婦女服用抗精神病藥物對姙娠結果之影響
論文名稱(英文) Investigation of Pregnancy Outcomes of Women Using Antipsychotic Drugs
校院名稱 臺北醫學大學
系所名稱(中) 藥學研究所
系所名稱(英) Graduate Institute of Pharmacy
學年度 97
學期 2
出版年 98
研究生(中文) 吳芳仁
學號 M301096024
學位類別 碩士
語文別 中文
口試日期 2009-06-22
論文頁數 88頁
口試委員 指導教授-許秀蘊
共同指導教授-林恆慶
委員-陳楚杰
委員-陳欽賢
委員-李信謙
關鍵字(中) 非典型抗精神病藥物
典型抗精神病藥物

娠結果
精神分裂症
全民健康保險研究資料庫
關鍵字(英) aypical antipsychotic
pregnancy outcomes
schizophrenia
typical antipsychotic
National Health Insurance Research database
學科別分類
中文摘要 研究背景:精神疾病在女性患者好發於育齡時期。根據最近研究報導顯示,孕婦服用抗精神病藥物有可能造成不良姙娠結果,包括新生兒早產、低出生體重、出生體重小於或大於姙娠年齡、以及增加孕婦剖腹產比率等。然而,懷孕婦女服用抗精神病藥物對姙娠結果之影響,特別是針對患有精神分裂症者,國內至今缺乏大型資料庫研究,因此難以確切提供醫療專業人員和孕婦服用抗精神病藥物之參考。
研究目的:本研究使用全國人口資料(nationwide population-based study),探討懷孕婦女服用抗精神病藥物,特別是患有精神分裂症孕婦,對於姙娠結果之影響。不良姙娠結果包括新生兒低出生體重、早產、小於姙娠年齡、大於姙娠年齡與產婦剖腹產計五項。
研究方法:本研究以1996年到2003年國家衛生研究院所提供之台灣全民健保研究資料庫(Taiwan National Health Insurance Research Dataset, NHIRD),門診處方及治療明細檔(適用85∼92年資料)(CD檔)、門診處方醫令明細檔(OO檔)、住院檔(DD檔)和承保資料檔(ID檔),串連內政部「2001-2003年的出生登記檔」;研究對象為患有精神分裂症(ICD-9-CM code 295, 排除295.7- schizoaffective disorder)之單胞胎產婦。統計方法使用Statistics Analysis System (SAS)統計軟體,在校正產婦個人、產婦配偶和新生兒特質等潛在干擾因子後,利用多變項羅吉斯迴歸 (multiple logistic regression analysis)分析懷孕婦女服用抗精神病藥物對姙娠結果之影響,結果以勝算比(adjusted odds ratio)、95%信賴區間(confidence interval)和 p-value <0.05作為統計上顯著差異。
研究結果:本研究對象為收錄於全民健保資料庫中,患有精神分裂症(ICD-9-CM code 295) 696名單胞胎產婦,以及隨機選取同期間資料庫中3,480位無精神分裂症之一般懷孕婦女。結果發現,接受典型抗精神病藥物治療產婦,相較於姙娠期間未接受抗精神病藥物治療產婦,其新生兒早產勝算比高達2.48 (crude OR=2.48, p<0.001);但是,不論校正前後,產婦於姙娠期間接受非典型抗精神病藥物治療之不良姙娠結果勝算比,相較於產婦姙娠期間未接受抗精神病藥物治療,在統計上並無顯著差異。另外,無精神分裂症之正常懷孕婦女,新生兒低出生體重之勝算比和小於姙娠年齡之勝算比,皆較精神分裂症懷孕婦女偏低,且呈現統計上顯著差異。
結論:本研究為迄今第一篇以全民健保資料庫,探討2001年至2003年期間,懷孕婦女服用抗精神病藥物對姙娠結果之影響。本研究發現,相較於姙娠期間未接受抗精神病藥物治療產婦,精神分裂症孕婦接受接受非典型抗精神病藥物治療,其不良姙娠結果並無統計上差異,但是精神分裂症孕婦服用典型抗精神病藥物,其新生兒早產危險性,在統計上呈現倍數增加。本研究的發現期盼有助於提供醫療專業人員和孕婦服用抗精神病藥物之諮詢參考。
英文摘要 Background: The age of onset of psychotic disorders is usually before or during the childbearing years. Several studies reported that women using antipsychotics may increase adverse pregnancy outcomes, particularly infant low birthweight (LBW), preterm birth, small for gestational age (SGA), large for gestational age (LGA) and maternal cesarean delivery during the past few years. However, the safety of antipsychotics on pregnancy outcome, especially for women with schizophrenia in Taiwan, remains unclear.

Objectives: The purpose of this nationwide population-based study was to compare the risk of adverse pregnancy outcomes between mothers with schizophrenia receiving antipsychotics during pregnancy and comparison subjects. The adverse pregnancy outcomes including preterm birth, LBW, LGA, SGA and maternal cesarean delivery.

Methods: This study linked two nationwide population-based datasets. The first dataset was obtained from the Taiwan National Health Insurance Research Dataset (NHIRD) and the second from the 2001-2003 birth certificate registry published by the Ministry of Interior. The Statistics Analysis System (SAS) statistical package was used to perform the analyses in this study. Multivariate logistic regression analysis was performed to explore the risk of adverse pregnancy outcomes between schizophrenia mothers and nonschizophrenia mothers after adjusting for the characteristics of mother, father and infant. The odds ratios (OR) and 95% confidence intervals (CI) for the estimated ORs were calculated. A two-sided p-value of <0.05 was considered statistically significant for this study.

Results: A total of 696 mothers with schizophrenia and singleton live births between January 1, 2001 and December 31, 2003 were included as the study group and 3,480 randomly selected pregnant women as a comparison group. As compared with maternal schizophrenia who did not receive antipsychotics during pregnancy, schizophrenia mothers who received typical antipsychotics during pregnancy had higher odds of preterm births (OR=2.48, 95% CI=1.51-4.08, p<0.001). Nevertheless, schizophrenia mothers who received atypical antipsychotics during pregnancy did not have higher odds of LBW infants, preterm births, SGA or LGA babies. And mothers with schizophrenia, whether receiving antipsychotics or not during pregnancy, had increased risks for delivering babies with LBW and SGA than the comparison cohort.

Conclusions: To the best of our knowledge, this is the first nationwide population-based study to investigate the risk of adverse pregnancy outcomes among mothers with schizophrenia receiving typical, atypical, and no antipsychotics during pregnancy and comparison subjects from the NHIRD database in Taiwan. This study indicate that comparing to mothers with schizophrenia who received no antipsychotics during pregnancy, there were no higher risks of adverse pregnancy outcomes observed among those receiving atypical agents. The findings of the current study may help clinicians understand the possible impact of antipsychotics on pregnancy outcome and indicate a better clinical option for optimal control of schizophrenic symptoms during pregnancy.
論文目次 目錄
致謝 I
中文摘要 III
Abstract VI
目錄 IX
圖目錄 X
表目錄 XI
第一章 緒論 1
第一節 研究背景 1
第二節 研究目的 2
第二章 文獻探討 3
第一節 精神分裂症 3
第二節 精神分裂症的流行病學 6
第三節 精神分裂症的抗精神病藥物治療 8
第四節 精神分裂症相關不良懷孕結果 16
第五節 抗精神病藥物相關之不良懷孕結果 18
第三章 研究方法 36
第一節 研究設計 36
第二節 研究架構 37
第三節 研究假說 39
第四節 研究對象 40
第五節 資料來源 42
第六節 研究變項與操作型定義 45
第七節 統計分析 48
第四章 研究結果 49
第一節 描述性分析 49
第二節 雙變項分析 56
第三節 多變項分析 62
第五章 討論 70
第一節 研究方法討論 70
第二節 研究結果討論 71
第三節 研究限制 78
第六章 結論與建議 79
第一節 結論 79
第二節 建議 79
參考文獻 82
附錄-本論文相關名稱縮寫表 88

圖目錄
圖1 研究架構 38
圖2 研究對象 41
圖3 精神分裂症產婦抗精神病藥物使用狀況 50
圖4 比較精神分裂症產婦接受典型或非典型抗精神病藥物不良姙娠結果之校正後勝算比 67

表目錄
表1 精神分裂症正、負向症狀 4
表2 台灣地區2001-2003年精神疾病患者統計結果-按性別及年齡 7
表3 Dopamin神經細胞之主要路徑與功能 9
表4 目前常用抗精神病藥物 10
表5 FDA懷孕期間用藥分級原則 12
表6 常用抗精神病藥物藥動學(pharmacokinetic) 13
表7 常用抗精神病藥物接受體-鍵結相對親和力 14
表8 常用抗精神病藥物副作用相對發生率(relative incidence)15
表9 Chlorpromazine、thioridazine和perphenazine相關不良姙娠結果報告 20
表10 2001-2008年haloperidol相關不良姙娠結果研究 24
表11 2001-2008年pimozide相關不良姙娠結果研究 25
表12 2001-2008年clozapine相關不良姙娠結果研究 28
表13 2001-2008年olanzapine相關不良姙娠結果研究 30
表14 2001-2008年quetiapine相關不良姙娠結果研究 32
表15 2001-2008年risperidone相關不良姙娠結果研究 33
表16 Risperidone藥品之新生兒畸形報告 34
表17 2001-2008年aripiprazole相關不良姙娠結果研究 35
表18 變項操作型定義 47
表19 精神分裂症產婦於懷孕期間服用抗精神病藥物之使用狀況 51
表20 無精神分裂症與精神分裂症產婦(不同藥物治療方式)之人口學特質分析 54
表21 卡方檢定無精神分裂症與精神分裂症產婦人口學特質之關聯性 57
表22 無精神分裂症與精神分裂症產婦不良姙娠結果之單變項羅吉斯迴歸分析 61
表23 無精神分裂症與精神分裂症產婦不良姙娠結果之多變項羅吉斯迴歸分析 66
表24 校正前後無精神分裂症與精神分裂症產婦不良姙娠結果之比較 69
表25 本研究結果與國內外相關研究的比較 77
參考文獻 孔繁鐘編譯(2007):DSM-IV-TR 精神疾病診斷準則手冊。 臺北:合記圖書出版社
沈武典(2007):21世紀臨床精神藥物學(第2版)。 臺北:合記圖書出版社。
張明永(2003)。全民健保精神分裂症住院病患精神科藥品使用狀態及其經濟效益評估。行政院衛生署九十年度委託研究計畫,DOH90-NH-010。
中央健康保險局,http://www.nhi.gov.tw/webdata/webdata/藥品給付規定,2009/06。
行政院衛生署衛生統計資訊網,http://www.doh.gov.tw/cht2006/統計室,2009/06。
國家衛生研究院,http://www.nhri.org.tw/全民健康保險研究資料庫,2009/06。
ACOG Practice Bulletin: Clinical management guidelines for obstetrician-gynecologists number 92, April 2008 (replaces practice bulletin number 87, November 2007). Use of psychiatric medications during pregnancy and lactation (2008). Obstet Gynecol, 111(4), 1001-1020.
Aichhorn, W., Whitworth, A. B., Weiss, E. M., & Marksteiner, J. (2006). Second-generation antipsychotics: is there evidence for sex differences in pharmacokinetic and adverse effect profiles? Drug Saf, 29(7), 587-598.
Aichhorn, W., Yazdi, K., Kralovec, K., Steiner, H., Whitworth, S., & Stuppaeck, C. (2008). Olanzapine plasma concentration in a newborn. J Psychopharmacol, 22(8), 923-924.
Allison, S. K. (2004). Psychotropic medication in pregnancy: ethical aspects and clinical management. J Perinat Neonatal Nurs, 18(3), 194-205.
Altshuler, L. L., Cohen, L., Szuba, M. P., Burt, V. K., Gitlin, M., & Mintz, J. (1996). Pharmacologic management of psychiatric illness during pregnancy: dilemmas and guidelines. Am J Psychiatry, 153(5), 592-606.
American Academy of Pediatrics (2000). Use of psychoactive medication during pregnancy and possible effects on the fetus and newborn. Committee on Drugs. Pediatrics, 105(4 Pt 1), 880-887.
Antipsychotic Agents. (2007) In: Drug Facts and Comparisons Pocket Version, 11ed, A Wolters Kluwer. pp.640-41.
Arora, M., & Praharaj, S. K. (2006). Meningocele and ankyloblepharon following in utero exposure to olanzapine. Eur Psychiatry, 21(5), 345-346.
Bennedsen, B. E. (1998). Adverse pregnancy outcome in schizophrenic women: occurrence and risk factors. Schizophr Res, 33(1-2), 1-26.
Bennedsen, B. E., Mortensen, P. B., Olesen, A. V., & Henriksen, T. B. (1999). Preterm birth and intra-uterine growth retardation among children of women with schizophrenia. Br J Psychiatry, 175, 239-245.
Bennedsen, B. E., Mortensen, P. B., Olesen, A. V., Henriksen, T. B., & Frydenberg, M. (2001). Obstetric complications in women with schizophrenia. Schizophr Res, 47(2-3), 167-175.
Biswasl, P. N., Wilton, L. V., Pearcel, G. L., Freemantle, S., & Shakir, S. A. (2001). The pharmacovigilance of olanzapine: results of a post-marketing surveillance study on 8858 patients in England. J Psychopharmacol, 15(4), 265-271.
Bjarnason, N. H., Rode, L., & Dalhoff, K. (2006). Fetal exposure to pimozide: a case report. J Reprod Med, 51(5), 443-444.
Briggs, G.G., Freeman, R.K. & Yaffe S.J. (Eds), (2005). Drugs in Pregnancy and Lactation: A Reference Guide to Fetal and Neonatal Risk. (7th ed.) Philadelphia (PA): Lippincott Williams & Wilkins.
Chien, I. C., Chou, Y. J., Lin, C. H., Bih, S. H., Chou, P., & Chang, H. J. (2004). Prevalence and incidence of schizophrenia among national health insurance enrollees in Taiwan, 1996-2001. Psychiatry Clin Neurosci, 58(6), 611-618.
Chien, I. C., Hsu, J. H., Bih, S. H., Lin, C. H., Chou, Y. J., Lee, C. H., et al. (2008). Prevalence, correlates, and disease patterns of antipsychotic use in Taiwan. Psychiatry Clin Neurosci, 62(6), 677-684.
Coppola, D., Russo, L. J., Kwarta, R. F., Jr., Varughese, R., & Schmider, J. (2007). Evaluating the postmarketing experience of risperidone use during pregnancy: pregnancy and neonatal outcomes. Drug Saf, 30(3), 247-264.
Dabbert, D., & Heinze, M. (2006). Follow-up of a pregnancy with risperidone microspheres. Pharmacopsychiatry, 39(6), 235.
Dervaux, A., Ichou, P., Pierron, G., Devianne, F., & Bavoux, F. (2007). Olanzapine exposure during pregnancy. Aust N Z J Psychiatry, 41(8), 706.
Diav-Citrin, O., Shechtman, S., Ornoy, S., Arnon, J., Schaefer, C., Garbis, H., et al. (2005). Safety of haloperidol and penfluridol in pregnancy: a multicenter, prospective, controlled study. J Clin Psychiatry, 66(3), 317-322.
Duran, A., Ugur, M. M., Turan, S., & Emul, M. (2008). Clozapine use in two women with schizophrenia during pregnancy. J Psychopharmacol, 22(1), 111-113.
Endow-Eyer, R.A., Mitchell, M.M., & Lacro, J.P., (2008). Schizophrenia. In: Koda-Kimble, Mary Anne (eds). Applied Therapeutics: the clinical use of drugs (9th ed.): Philadelphia: Lippincott Williams & Wilkins, 78:1-33.
Essock, S. M. (2002). Editor's introduction: antipsychotic prescribing practices. Schizophr Bull, 28(1), 1-4.
Food and Drug Administration. Federal Register 1980;44:37434-67
Fowden AL. Growth and metabolism. In: Harding R, Bocking AD (eds). Fetal Growth and Development. Cambridge University Press: Cambridge, UK, 2001; 44-68.
Freedman, R. (2003). Schizophrenia. N Engl J Med, 349(18), 1738-1749.
Friedman, S. H., & Rosenthal, M. B. (2003). Treatment of perinatal delusional disorder: a case report. Int J Psychiatry Med, 33(4), 391-394.
Gentile, S. (2006). Quetiapine-fluvoxamine combination during pregnancy and while breastfeeding. Arch Womens Ment Health, 9(3), 158-159.
Gentile, S. (2008). Antipsychotic Therapy During Early and Late Pregnancy. A Systematic Review. Schizophr Bull. Sep 11
Godet, P. F., & Marie-Cardine, M. (1991). [Neuroleptics, schizophrenia and pregnancy. Epidemiological and teratologic study]. Encephale, 17(6), 543-547.
Goldstein, D. J., Corbin, L. A., & Fung, M. C. (2000). Olanzapine-exposed pregnancies and lactation: early experience. J Clin Psychopharmacol, 20(4), 399-403.
Grover, S., & Avasthi, A. (2004). Risperidone in pregnancy: a case of oligohydramnios [in German]. J Psychiatry (7), 56–57.
Gupta, N., & Grover, S. (2004). Safety of clozapine in 2 successive pregnancies. Can J Psychiatry, 49(12), 863.
Hall G., (1963). A case of phocomelia of the upper limbs. Med J Aust (1), 449-50
Hauser, L. A. (1985). Pregnancy and psychiatric drugs. Hosp Community Psychiatry, 36(8), 817-818.
Jablensky, A. V., Morgan, V., Zubrick, S. R., Bower, C., & Yellachich, L. A. (2005). Pregnancy, delivery, and neonatal complications in a population cohort of women with schizophrenia and major affective disorders. Am J Psychiatry, 162(1), 79-91.
Karakula, H., Szajer, K., Rpila, B., Grzywa, A., & Chuchra, M. (2004). Clozapine and pregnancy--a case history. Pharmacopsychiatry, 37(6), 303-304.
Kulkarni, J., McCauley-Elsom, K., Marston, N., Gilbert, H., Gurvich, C., de Castella, A., et al. (2008). Preliminary findings from the National Register of Antipsychotic Medication in Pregnancy. Aust N Z J Psychiatry, 42(1), 38-44.
Lang, H. C., & Su, T. P. (2004). The cost of schizophrenia treatment in Taiwan. Psychiatr Serv, 55(8), 928-930.
Lawrence, E. J. (2006). Part 1: a matter of size. Evaluating the growth-restricted neonate. Adv Neonatal Care, 6(6), 313-322.
Lawrence, E. J. (2007). A matter of size: Part 2. Evaluating the large-for-gestational-age neonate. Adv Neonatal Care, 7(4), 187-197.
Lee, A., Giesbrecht, E., Dunn, E., & Ho, S. (2004). Excretion of quetiapine in breast milk. Am J Psychiatry, 161(9), 1715-1716.
Lehman, A. F., Lieberman, J. A., Dixon, L. B., McGlashan, T. H., Miller, A. L., Perkins, D. O., et al. (2004). Practice guideline for the treatment of patients with schizophrenia, second edition. Am J Psychiatry, 161(2 Suppl), 1-56.
Lin, H. C., Tang, C. H., & Lee, H. C. (2008). Association Between Paternal Schizophrenia and Low Birthweight: A Nationwide Population-Based Study. Schizophr Bull, 1-7
Mauri, M. C., Bravin, S., Bitetto, A., Rudelli, R., & Invernizzi, G. (1996). A risk-benefit assessment of sulpiride in the treatment of schizophrenia. Drug Saf, 14(5), 288-298.
McCauley-Elsom, K., & Kulkarni, J. (2007). Managing psychosis in pregnancy. Aust N Z J Psychiatry, 41(3), 289-292.
McKenna, K., Koren, G., Tetelbaum, M., Wilton, L., Shakir, S., Diav-Citrin, O., et al. (2005). Pregnancy outcome of women using atypical antipsychotic drugs: a prospective comparative study. J Clin Psychiatry, 66(4), 444-449
Mendhekar, D. N., War, L., Sharma, J. B., & Jiloha, R. C. (2002). Olanzapine and pregnancy. Pharmacopsychiatry, 35(3), 122-123.
Mendhekar, D. N., Sharma, J. B., Srivastava, P. K., & War, L. (2003). Clozapine and pregnancy. J Clin Psychiatry, 64(7), 850.
Mendhekar, D. N., Sunder, K. R., & Andrade, C. (2006). Aripiprazole use in a pregnant schizoaffective woman. Bipolar Disord, 8(3), 299-300.
Mendhekar, D.N., Sharma, J.B., & Srilakshmi P. (2006). Use of aripiprazole during late pregnancy in a woman with psychotic illness [letter]. Ann Pharmacother. 40:575.
Mendhekar, D. N. (2007). Possible delayed speech acquisition with clozapine therapy during pregnancy and lactation. J Neuropsychiatry Clin Neurosci, 19(2), 196-197.
Mervak, B., Collins, J., & Valenstein, M. (2008). Case report of aripiprazole usage during pregnancy. Arch Womens Ment Health, 11(3), 249-250.
Miller, L. J. (1997). Sexuality, reproduction, and family planning in women with schizophrenia. Schizophr Bull, 23(4), 623-635.
Miyamoto, S., Duncan, G. E., Marx, C. E., & Lieberman, J. A. (2005). Treatments for schizophrenia: a critical review of pharmacology and mechanisms of action of antipsychotic drugs. Mol Psychiatry, 10(1), 79-104.
Moya, F., & Thorndike, V. (1962). Passage of drugs across the placenta. Am J Obstet Gynecol, 84, 1778-1798.
Nagy, A., Tenyi, T., Lenard, K., Herold, R., Wilhelm, F., & Trixler, M. (2001). [Olanzapine and pregnancy]. Orv Hetil, 142(3), 137-138.
Newham, J. J., Thomas, S. H., MacRitchie, K., McElhatton, P. R., & McAllister-Williams, R. H. (2008). Birth weight of infants after maternal exposure to typical and atypical antipsychotics: prospective comparison study. Br J Psychiatry, 192(5), 333-337.
Newport, D. J., Calamaras, M. R., DeVane, C. L., Donovan, J., Beach, A. J., Winn, S., et al. (2007). Atypical antipsychotic administration during late pregnancy: placental passage and obstetrical outcomes. Am J Psychiatry, 164(8), 1214-1220.
Nguyen, H. N., & Lalonde, P. (2003). [Clozapine and pregnancy]. Encephale, 29(2), 119-124.
Nilsson, E., Lichtenstein, P., Cnattingius, S., Murray, R. M., & Hultman, C. M. (2002). Women with schizophrenia: pregnancy outcome and infant death among their offspring. Schizophr Res, 58(2-3), 221-229.
Nilsson, E., Hultman, C. M., Cnattingius, S., Olausson, P. O., Bjork, C., & Lichtenstein, P. (2008). Schizophrenia and offspring's risk for adverse pregnancy outcomes and infant death. Br J Psychiatry, 193(4), 311-315.
Patton, S. W., Misri, S., Corral, M. R., Perry, K. F., & Kuan, A. J. (2002). Antipsychotic medication during pregnancy and lactation in women with schizophrenia: evaluating the risk. Can J Psychiatry, 47(10), 959-965.
Ratnayake, T., & Libretto, S. E. (2002). No complications with risperidone treatment before and throughout pregnancy and during the nursing period. J Clin Psychiatry, 63(1), 76-77.
Ratnayake, T., & Libretto, S. E. (2002). No complications with risperidone treatment before and throughout pregnancy and during the nursing period. J Clin Psychiatry, 63(1), 76-77.
Reis, M., & Kallen, B. (2008). Maternal use of antipsychotics in early pregnancy and delivery outcome. J Clin Psychopharmacol, 28(3), 279-288.
Rumeau-Rouquette, C., Goujard, J., & Huel, G. (1977). Possible teratogenic effect of phenothiazines in human beings. Teratology, 15(1), 57-64.
Sacker, A., Done, D.J. & Crow, T.J. (1996). Obstetrics complications in children born of parents with schizophrenia: A meta-analysis of case-control studies. Psychological Medicine, 26 (2), 279–287.
Saha, S., Chant, D., Welham, J., & McGrath, J. (2005). A systematic review of the prevalence of schizophrenia. PLoS Med, 2(5): 413-33.
Scanlan, F. J. (1972). The use of thioridazine (Melleril) during the first trimester. Med J Aust, 1(24), 1271-1272.
Schizophrenia and other psychotic disorders. (2000) In: Diagnostic and statistical manual of mental disorders, 4th edition, text revision. Washington, D.C., American Psychiatric Association, pp.297-319
Schneid-Kofman, N., Sheiner, E., & Levy, A. (2008). Psychiatric illness and adverse pregnancy outcome. Int J Gynaecol Obstet, 101(1), 53-56.
Seeman, M. V. (2004). Gender differences in the prescribing of antipsychotic drugs. Am J Psychiatry, 161(8), 1324-1333.
Shen, W.W. (1999). A history of antipsychotic drug development. Comprehensive Psychiatry, 40(6):407-414.
Slone, D., Siskind, V., Heinonen, O. P., Monson, R. R., Kaufman, D. W., & Shapiro, S. (1977). Antenatal exposure to the phenothiazines in relation to congenital malformations, perinatal mortality rate, birth weight, and intelligence quotient score. Am J Obstet Gynecol, 128(5), 486-488.
Spyropoulou, A. C., Zervas, I. M., & Soldatos, C. R. (2006). Hip dysplasia following a case of olanzapine exposed pregnancy: a questionable association. Arch Womens Ment Health, 9(4), 219-222.
Taylor, T. M., O'Toole, M. S., Ohlsen, R. I., Walters, J., & Pilowsky, L. S. (2003). Safety of quetiapine during pregnancy. Am J Psychiatry, 160(3), 588-589.
Tenyi, T., Trixler, M., & Keresztes, Z. (2002). Quetiapine and pregnancy. Am J Psychiatry, 159(4), 674.
Tucker, J., & McGuire, W. (2004). Epidemiology of preterm birth. BMJ, 329(7467), 675-678.
Twaites, B. R., Wilton, L. V., & Shakir, S. A. (2007). The safety of quetiapine: results of a post-marketing surveillance study on 1728 patients in England. J Psychopharmacol, 21(4), 392-399.
Vemuri, M. P., & Rasgon, N. L. (2007). A case of olanzapine-induced gestational diabetes mellitus in the absence of weight gain. J Clin Psychiatry, 68(12), 1989.
Vince, D. J. (1969). Congenital malformations following phenothiazine administration during pregnancy. Ca Med Assoc J, 100(4), 223.
Yeshayahu, Y. (2007). The use of olanzapine in pregnancy and congenital cardiac and musculoskeletal abnormalities. Am J Psychiatry, 164(11), 1759-1760.
Yogev, Y., Ben-Haroush, A., & Kaplan, B. (2002). Maternal clozapine treatment and decreased fetal heart rate variability. Int J Gynaecol Obstet, 79(3), 259-260.
Yonkers, K. & Little, B. (Eds.). (2001). Management of psychiatric disorders in pregnancy. London: Arnold; pp 188-212

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系統識別號 U0007-1108200912371200
論文名稱(中文) 應用全民健保資料庫研究懷孕期婦女使用抗憂鬱藥對胎兒的影響
論文名稱(英文) Neonatal Outcomes of Prenatal Antidepressant Exposure: A National Health Insurance Research Database Study
校院名稱 臺北醫學大學
系所名稱(中) 藥學研究所
系所名稱(英) Graduate Institute of Pharmacy
學年度 97
學期 2
出版年 98
研究生(中文) 林信成
學號 M301096026
學位類別 碩士
語文別 中文
口試日期 2009-07-15
論文頁數 69頁
口試委員 委員-曾啟瑞
委員-沈武典
委員-劉亞平
指導教授-吳姿樺
共同指導教授-林恆慶
關鍵字(中) 憂鬱症
孕婦
新生兒健康風險
健保資料庫
抗憂鬱劑
關鍵字(英) antidepressants
birth outcome
depression
neonate
pregnant women
學科別分類
中文摘要 女性憂鬱症的發生率是男性的兩倍。且好發的年齡層在25歲到44歲之間;又不論是憂鬱症本身或是用於治療憂鬱症藥物皆有報導可能與造成胎兒或新生兒健康指標不良的風險有關。為了能提出更有力的藥物流行病學研究實證以期能了解懷孕期服用抗憂鬱藥的用藥安全問題,本研究應用台灣全民健保資料庫探討懷孕期婦女使用抗憂鬱藥對胎兒的影響。首先利用民國2001-2003年健保資料庫並串聯出生檔,剔除多胞胎新生兒後及懷孕期間使用抗憂鬱藥累計未達14天之病患,取得樣本數468,471人;之後比較懷孕期使用抗憂鬱藥(n=1,713)孕婦分娩胎兒及懷孕期間未使用抗憂鬱藥孕婦分娩新生兒(466,758)健康指標。並設定依變項為胎兒之早產與否(出生周數<37週)、出生體重(<2,500公克)及出生體重小於妊娠年齡(SGA),自變項為孕婦於懷孕期間使用抗憂鬱。繼而以頻率(百分比)描述樣本特性,其次再透過卡方檢定(Chi-square)檢定關聯性最後利用羅吉氏迴歸(logistic regression)分析不同成分抗憂鬱藥的勝算比。研究結果發現懷孕期間有使用trazodone抗憂鬱藥所分娩胎兒相較於孕婦於懷孕期間沒有使用抗憂鬱藥所分娩胎兒三項新生兒出生指標:早產(odds ratio=1.243;95% CI=1.151-1.34)、出生體重過輕(odds ratio=1.157; 95% CI=1.059-1.263)及出生體重小於小於妊娠年齡) (odds ratio=1.106; 95%CI=1.041-1.175)的風險;而使用paroxetine的孕婦對其所分娩的胎兒具出生體重過輕及出生體重小於妊娠年齡兩指標之關聯性,且均達到統計顯顯著之差異。本研究推論懷孕期間臨床上應避免抗憂鬱藥品投予懷孕婦女,若於病症無其他更適合控制病情之藥品選擇時,應提醒注意定期追蹤胎兒成長與健康情形。
英文摘要 Objectives: The incidence of depressive disorders in women is two times higher compared to that in men. The prevalence in women is the highest during their childbearing years. Maternal depression is associated with perinatal risk related to physiological sequelae or inadequate obstetrical care. Therefore, to clarify the drug-related problems for the maternal depression therapy with providing pharmacoepidemiological evidence, we conducted this study and used a nationwide database released by the Taiwa National Health Research Institute to link to the birth records. The study cohort consisted of the pregnant women who had delivered during 2001-2003 and met the research inclusion criteria (n=468,471). The risks of preterm birth, low birth weight, small for gestationa age of exposed pregnant women (n=1,713) were compared with those neonatal outcomes of non-exposed women (466,758). The statistical significance of the results showed that the neonate outcomes of maternal exposures to trazodone had high risks of preterm birth (odds ratio=1.243; 95% CI= 1.151-1.34), low birth weight (odds ratio=1.157; 95% CI=1.059-1.263), and small for gestational age (odds ratio=1.106; 95% CI=1.041-1.175). Exposures to paroxetine had increaesd risks of low birth weight (odds ratio=2.163; 95% CI=1.077-4.343), small for gestationa age (odds ratio=1.816; 95% CI=1.078-3.058). The result of this study suggest that the uses of antidepressants particularly trazodone and paroxetine during pregnant women need to be careful and long-term monitoing for those neonatal health is necessary.
論文目次 目 錄
第一章 緒論----------------------------------------------- 1
第一節 研究背景------------------------------------------ 2
第二節 研究目的------------------------------------------ 3
第三節 全民健康保險學術研究資料庫------------------------- 3第二章 文獻探討------------------------------------------- 5
第一節 憂鬱症症狀---------------------------------------- 6
第二節 憂鬱症的診斷評估及防治----------------------------- 6
第三節 憂鬱症的治療方法---------------------------------- 7
第四節 常用抗憂鬱藥物------------------------------------- 7
第三章 研究方法-------------------------------------------10
第一節 研究架構 -----------------------------------------11
第二節 全民健康保險學術研究資料庫與研究樣本來源----------11
第三節 研究變項與操作型定義------------------------------12
第三節 資料處理及分析方法--------------------------------13
第四章 研究結果------------------------------------------15
第一節 孕婦使用之抗憂鬱藥與新生兒早產關聯性及勝算比------17
第二節 孕婦使用之抗憂鬱藥與新生兒出生體重過輕關聯性------21
第三節 孕婦使用抗憂鬱藥與胎兒出生體重小於妊娠年齡關聯性及勝
算比----------------------------------------------27
第五章 討論----------------------------------------------32
第六章 結論----------------------------------------------35圖表-----------------------------------------------------57
參考文獻-------------------------------------------------65
參考文獻 1.Burden of Mental and Behavioural Disorders. 2001; http://www.who.int/whr2001/2001/main/en/chapter2/index.htm.
2.The world health report 2001 - Mental Health: New Understanding, New Hope. 2001; 2008-2010-2019. Available at: http://www.who.int/whr/2001/en/index.html.
3.Andrade L, Caraveo-Anduaga JJ, Berglund P, et al. The epidemiology of major depressive episodes: results from the International Consortium of Psychiatric Epidemiology (ICPE) Surveys. Int J Methods Psychiatr Res. 2003;12(1):3-21.
4.Kessler RC, Berglund P, Demler O, et al. The epidemiology of major depressive disorder: results from the National Comorbidity Survey Replication (NCS-R). JAMA. Jun 18 2003;289(23):3095-3105.
5.Kessler RC, Berglund P, Demler O, Jin R, Merikangas KR, Walters EE. Lifetime prevalence and age-of-onset distributions of DSM-IV disorders in the National Comorbidity Survey Replication. Arch Gen Psychiatry. Jun 2005;62(6):593-602.
6.Murphy JM, Laird NM, Monson RR, Sobol AM, Leighton AH. A 40-year perspective on the prevalence of depression: the Stirling County Study. Arch Gen Psychiatry. Mar 2000;57(3):209-215.
7.Kuehner C. Gender differences in unipolar depression: an update of epidemiological findings and possible explanations. Acta Psychiatr Scand. Sep 2003;108(3):163-174.
8.Bennett HA, Einarson A, Taddio A, Koren G, Einarson TR. Depression during Pregnancy : Overview of Clinical Factors. Clin Drug Investig. 2004;24(3):157-179.
9.Kessler RC, McGonagle KA, Zhao S, et al. Lifetime and 12-month prevalence of DSM-III-R psychiatric disorders in the United States. Results from the National Comorbidity Survey. Arch Gen Psychiatry. Jan 1994;51(1):8-19.
10.Weissman MM, Olfson M. Depression in women: implications for health care research. Science. Aug 11 1995;269(5225):799-801.
11.Public Health Advisory Paroxetine. http://www.fda.gov/Drugs/DrugSafety/PublicHealthAdvisories/ucm051731.htm. .
12.Chambers CD, Johnson KA, Dick LM, Felix RJ, Jones KL. Birth outcomes in pregnant women taking fluoxetine. N Engl J Med. Oct 3 1996;335(14):1010-1015.
13.Kulin NA, Pastuszak A, Sage SR, et al. Pregnancy outcome following maternal use of the new selective serotonin reuptake inhibitors: a prospective controlled multicenter study. JAMA. Feb 25 1998;279(8):609-610.
14.Oberlander TF WW, Misri S, Aghajanian J, Hertzman C. . Neonatal outcomes after prenatal exposure to selective serotonin reuptake inhibitor antidepressants and maternal depression using population-based linked health data. . Arch Gen Psychiatry.;63(8):898-908.
15.Depression. http://www.nimh.nih.gov/health/publications/depression/nimhdepression.pdf.
16.Hays RD, Wells KB, Sherbourne CD, Rogers W, Spritzer K. Functioning and well-being outcomes of patients with depression compared with chronic general medical illnesses. Arch Gen Psychiatry. Jan 1995;52(1):11-19.
17.邱南英. 使用抗憂鬱劑治療憂鬱症的新趨勢. 彰化市2003.
18.邱南英. 憂鬱症. 財團法人彰化基督教醫院. 彰化市2003.
19.漢氏憂鬱症量表. http://www.health.gov.tw/Portals/0/%E9%86%AB%E8%AD%B7%E7%AE%A1%E7%90%86%E8%99%95/blue_3.doc.
20.WW. S. Clinical Psychopharmacology for the 21 century (in Mandarin). Clinical Psychopharmacology for the 21 century (in Mandarin). Taipei: Ho-Chi Publishing Company; 2007:353-385.
21.New federal guidelines seek to help primary care providers recognize and treat depression. Agency for Health Care Policy and Research (AHCPR) of the Department of Human Services. Hosp Community Psychiatry. Jun 1993;44(6):598.
22.Practice guideline for the treatment of patients with major depressive disorder (revision). American Psychiatric Association. Am J Psychiatry. Apr 2000;157(4 Suppl):1-45.
23.Thorpe L, Whitney DK, Kutcher SP, Kennedy SH. Clinical guidelines for the treatment of depressive disorders. VI. Special populations. Can J Psychiatry. Jun 2001;46 Suppl 1:63S-76S.
24.Rivett B. Modern Problems of Pharmacopsychiatry. Vol 9. 76-86 ed. Montreal, Quebec1973.
25.Troyer WA, Pereira GR, Lannon RA, Belik J, Yoder MC. Association of maternal lithium exposure and premature delivery. J Perinatol. Mar-Apr 1993;13(2):123-127.
26.Ericson A KB, Wilholm B-E. Delivery outcome after the use of antidepressants in early pregnancy. Eur J Clin Pharmacol 1999;7:503-508.
27.Simon GE, Cunningham ML, Davis RL. Outcomes of prenatal antidepressant exposure. Am J Psychiatry. Dec 2002;159(12):2055-2061.
28.Casper RC, Fleisher BE, Lee-Ancajas JC, et al. Follow-up of children of depressed mothers exposed or not exposed to antidepressant drugs during pregnancy. J Pediatr. Apr 2003;142(4):402-408.
29.Hendrick V, Smith LM, Suri R, Hwang S, Haynes D, Altshuler L. Birth outcomes after prenatal exposure to antidepressant medication. Am J Obstet Gynecol. Mar 2003;188(3):812-815.
30.Sivojelezova A, Shuhaiber S, Sarkissian L, Einarson A, Koren G. Citalopram use in pregnancy: prospective comparative evaluation of pregnancy and fetal outcome. Am J Obstet Gynecol. Dec 2005;193(6):2004-2009.
31.Malm H, Klaukka T, Neuvonen PJ. Risks associated with selective serotonin reuptake inhibitors in pregnancy. Obstet Gynecol. Dec 2005;106(6):1289-1296.
32.Einarson A, Choi J, Einarson TR, Koren G. Incidence of major malformations in infants following antidepressant exposure in pregnancy: results of a large prospective cohort study. Can J Psychiatry. Apr 2009;54(4):242-246.
33.Kallen K, Olausson PO. Neonatal mortality for low-risk women by method of delivery. Birth. Mar 2007;34(1):99-100; author reply 101-102.
34.Heikkinen T, Ekblad U, Kero P, Ekblad S, Laine K. Citalopram in pregnancy and lactation. Clin Pharmacol Ther. Aug 2002;72(2):184-191.
35.Louik C, Lin AE, Werler MM, Hernandez-Diaz S, Mitchell AA. First-trimester use of selective serotonin-reuptake inhibitors and the risk of birth defects. N Engl J Med. Jun 28 2007;356(26):2675-2683.
36.Greene MF. Teratogenicity of SSRIs--serious concern or much ado about little? N Engl J Med. Jun 28 2007;356(26):2732-2733.
37.Potts AL, Young KL, Carter BS, Shenai JP. Necrotizing enterocolitis associated with in utero and breast milk exposure to the selective serotonin reuptake inhibitor, escitalopram. J Perinatol. Feb 2007;27(2):120-122.
38.Goldstein DJ, Sundell KL, Corbin LA. Birth outcomes in pregnant women taking fluoxetine. N Engl J Med. Mar 20 1997;336(12):872-873; author reply 873.
39.ACOG Practice Bulletin: Clinical management guidelines for obstetrician-gynecologists number 92, April 2008 (replaces practice bulletin number 87, November 2007). Use of psychiatric medications during pregnancy and lactation. Obstet Gynecol. Apr 2008;111(4):1001-1020.
40.Alwan S, Reefhuis J, Rasmussen SA, Olney RS, Friedman JM. Use of selective serotonin-reuptake inhibitors in pregnancy and the risk of birth defects. N Engl J Med. Jun 28 2007;356(26):2684-2692.
41.Einarson A, Bonari L, Voyer-Lavigne S, et al. A multicentre prospective controlled study to determine the safety of trazodone and nefazodone use during pregnancy. Can J Psychiatry. Mar 2003;48(2):106-110.
42.Anon. Pregnancy categories for prescription drugs. Food and Drug Administration Drug Bulletin. 1982;12:24-25.



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系統識別號 U0007-1202200913104300
論文名稱(中文) 紫草素促傷口癒合之離體細胞培養以及活體動物試驗:SMAD 蛋白訊息傳遞路徑在紫草素誘發前膠原蛋白生成中所扮演之角色
論文名稱(英文) Effect of shikonin on promoting wound healing in vitro and in vivo : the role of SMAD signaling in shikonin induced procollagen formation
校院名稱 臺北醫學大學
系所名稱(中) 藥學研究所
系所名稱(英) Graduate Institute of Pharmacy
學年度 97
學期 1
出版年 98
研究生(中文) 楊若珺
學號 M301095005
學位類別 碩士
語文別 中文
口試日期 2009-01-05
論文頁數 85頁
口試委員 委員-顏茂雄
委員-王靜瓊
指導教授-鄭幼文
關鍵字(中) 紫草素
皮膚纖維母細胞
SMAD路徑
創傷癒合
關鍵字(英) shikonin
skin fibroblast
SMAD pathway
wound healing
學科別分類
中文摘要 皮膚是人體最大的器官,由多層上皮組織構成,覆蓋於整個人
體,形成屏障保護人體免於外來致病原。是故皮膚對人體而言相當重
要。當人體受傷時,會引發一連串的表皮創傷癒合過程,以修復並治
癒傷口,進行真皮與上皮組織的再建構,為人體的自然修復過程。通
常此過程可分為三階段互相重疊的時期,包括發炎反應、組織增生和
組織重建期。
紫草素是由紫草(Lithospermum euchroma, Boraginaceae)之根萃
取出的主要成份。在古代中國與日本入藥為「紫雲膏」來治療刀傷與
燒燙傷。許多文獻指出,紫草素具有許多生理活性,包括抗發炎、抗
菌、細胞凋亡、抗血管新生、抗腫瘤及促肉芽組織增生等。
肉芽組織生成與膠原蛋白堆積,為創傷癒合過程的組織增生期
中,至為重要的關鍵步驟。而人類纖維母細胞為肉芽組織和生成膠原
蛋白的主要細胞。在本文裡,我們分別透過活體動物實驗及離體試
驗,探討紫草素在促進創傷癒合過程中,對於人類皮膚纖維母細胞的
影響。
在活體動物實驗中,我們利用皮膚切片法,在小鼠背部創造出直
徑6 mm 的傷口。每天局部給予紫草素、bFGF(basic Fibroblast growth
factor)當正向控制組,PEG(Polyethylene glycol)為對照組。給藥期間為14 天,每天觀察測量傷口並記錄面積。由實驗結果發現,給予bFGF正向控制劑的組別擁有最顯著的癒合效果,另外,給予0.1%紫草素
的組別相較於對照組,促進傷口癒合的能力亦明顯較高,代表紫草素
參與了傷口癒合的過程。
為了進一步研究紫草素促進創傷癒合的機轉,我們選用人類纖維
母細胞細胞株(CCD-966SK)作為實驗細胞。在離體細胞培養實驗裡,
紫草素能夠誘發人類纖維母細胞的細胞遷移現象及ERK1/2 蛋白質
(Extracellular signal-regulated kinase)磷酸化,且呈現濃度相關。ERK
磷酸化通常與細胞增生反應有關。此外,紫草素亦能誘發MMP-1
(Matrix Metalloprotease-1)的蛋白質表現及抑制PAI-1(Plasminogen
activator inhibitor type I)蛋白質表現,也呈現濃度相關性。MMP-1 與
PAI-1 同時在ECM(Extracellular matrix)降解、纖維蛋白溶解和細胞遷
移中扮演重要的角色。另一方面,近年來SMAD3 路徑被認為是調控
創傷癒合的路徑之一,在紫草素的作用下,TGF-beta /SMAD 路徑中
的分子,SMAD3(Small mother against decapentaplegic)和procollagen
也會在人類皮膚纖維母細胞中被誘發出來。
綜合以上所述,從實驗結果推測,紫草素透過ERK 的磷酸化、
procollagen、PAI-1 和SMAD 的蛋白質表現,能加速創傷癒合,而
MMP-1 亦參與其中。MMP-1 在創傷癒合過程中的定位仍有所爭議,但也有報導指出,MMP-1 的誘發與創傷癒合有關。因此我們認為,紫草素在活體動物實驗與離體細胞試驗中,會透過SMAD 訊息傳遞
路徑,促使傷口癒合。而這些結果也提供了我們更多資訊,助於發展藥理臨床上創傷治療的應用。
英文摘要 Skin is the largest organ of our bodies, and made up of multiple
layers of epithelial tissues that cover the bodies protect us from the
external pathogens, also considered is one of the most important parts in
human organs. While an individual injuries, a set of events called
cutaneous wound healing will take place to repair the damage. It is a
natural process of regenerating dermal and epidermal tissue, and
categorized into several overlapping steps: the inflammatory, proliferative,
and maturation phases.
Shikonin is a major components isolated from the root of
Lithospermum euchroma (Boraginaceae). It was used as an ointment
called SHIUNKO for treating carbuncles and burns in China and Japan
for a long time. Many studies showed that shikonin demonstrated many
biological effects such as anti-inflammatory, antimicrobial, apoptotic,
anti-angiogenesis, anti-tumor, and proliferation of granulation tissue.
Since the granulation tissue formation and collagen deposition are
critical events of the proliferation phase of wound healing. In this study,
we investigated the effects of shikonin on human skin fibroblast on
promoting wound healing in vitro and in vivo.
In in vivo study, mice were created a 6 mm diameter wound on back
by biopsy punch. Topical treating with shikonin、bFGF (basic Fibroblast
growth factor)(positive control) and PEG(Polyethylene glycol)(negative
control) for 14 days, then measured and observed the wound area and
recorded everyday. Our data suggested bFGF treated groups showed
largely healing promoted effect and taken as positive control, 0.1%
shikonin also significantly promoted wound healing compared with
control mice. This data indicated the involvement of shikonin in wound
healing.
In order to investigate the mechanism of shikonin in promoting the
wound healing, we use human skin fibroblast cell culture line
(CCD-966SK) in our study. In in vitro cell culture experiments, we found
shikonin could concentration-dependently induce cell migration and
ERK1/2(Extracellular signal-regulated kinase) phosphorylation in human
skin fibroblasts. ERK phosphorylation is general accepted by contributing
to cell proliferation. In addition, shikonin also concentratio-dependently
increased MMP-1(Matrix metalloproteinase-1) induction, and decrease
PAI-1(Plasminogen activator inhibitor type I) inhibition. Both MMP-1
and PAI-1 play an important role for ECM (Extracellular matrix)
degradation, fibrinolysis and cell migration. On the other hand, the
molecule in TGF-beta/SMAD pathway, SMAD3 (Small mother against
decapentaplegic), and procollagen were all increased in shikonin treated
skin fibroblast cells. SMAD3 pathway is recognized in controlling the
healing pathway in recent years.
Take together, our data indicated, shikonin could accelerate wound
healing by promoting ERK phosphorylation and procollagen、PAI-I and SMAD protein induction, MMP-1 also involve in this effect. Although the role of MMP-1 in wound healing is controversial, but still have reports suggested the increase of MMP-1 is involved in the pathway. We concluded, shikonin can promote wound healing both in vitro and in vivo through SMAD signaling pathway, and this finding can provide the information for pharmacological use in the clinical care of wound healing.
論文目次 目錄
目錄 I
中文摘要 IV
Abstract VII
縮寫(Abbreviation) X

第一章 緒論 1
第一節 皮膚的構造 1
1-1.表皮層 1
1-2.真皮層 2
1-3.皮下組織 3
第二節 皮膚纖維母細胞(human skin fibroblast) 3
第三節 膠原蛋白(Collagen) 4
第四節 表皮創傷癒合(Cutaneous wound healing) 5
4-1.第一期:發炎反應期(Inflammatory phase) 5
4-2.第二期:組織增生期(Proliferative phase) 8
4-3.第三期:組織重建期(Maturation and remodeling phase) 11
第五節 紫草素簡介 12
5-1.紫草素背景 12
5-2.紫草素的藥理活性 13
第六節 MAPK 路徑與其調節作用Mitogen-activated protein kinase pathway 15
第七節 MMP-1基質金屬蛋白酵素(Matrix metalloproteinases, MMPs) 16
第八節 PAI-1纖維蛋白溶酶原活化抑制劑第一型 (Plasminogen Activator Inhibitor type I) 17
8-1.PAI-1背景簡介 17
8-2.纖維蛋白溶解系統 18
8-3.PAI-1的生理活性 19
第九節 TGF-β/SMAD 路徑(TGF-β/SMAD pathway) 19
9-1.TGF-β(Transforming growth factor- beta)轉化生長因子beta 19
9-2.SMAD(Small Mothers Against Decapentaplegic) 20
9-3.TGF-β/SMAD pathway(TGF-β/SMAD 路徑) 21
第十節 研究動機 22
第二章 實驗材料與方法 23
第一節 實驗材料 23
1-1.實驗藥品 23
1-2.套組試劑(Kit) 25
1-3.抗體(antibody) 25
1-4.耗材 26
1-5.實驗用儀器 26
第二節 實驗方法 27
2-1.活體動物創傷癒合試驗(Wound healing in vivo assay) 27
2-2.人類皮膚纖維母細胞(Human skin fibroblast)細胞株的培養 28
2-3.人類皮膚纖維母細胞(Human skin fibroblast)細胞株的繼代培養 28
2-4.細胞存活率與生長測定(MTT assay) 28
2-5.離體創傷癒合試驗(Wound healing in vitro assay) 30
2-6.西方墨點法 30
2-7.統計分析 32
第三章 實驗結果 33
第一節 探討Shikonin加速小鼠創傷癒合之現象 33
第二節 探討Shikonin對人類皮膚纖維母細胞的存活率與增生實驗 34
第三節 探討Shikonin誘發人類皮膚纖維母細胞磷酸化ERK1/2蛋白質的表現 34
第四節 探討Shikonin誘發人類受傷皮膚纖維母細胞的細胞增生與遷移現象 35
第五節 探討Shikonin誘發受傷人類皮膚纖維母細胞磷酸化ERK1/2蛋白質的表現 38
第六節 探討Shikonin抑制人類皮膚纖維母細胞PAI-1蛋白質的表現 39
第七節 探討Shikonin誘發人類皮膚纖維母細胞Procollagen type I蛋白質的表現 40
第八節 探討Shikonin誘發受傷人類皮膚纖維母細胞Procollagen type I蛋白質的表現 41
第九節 探討Shikonin誘發人類皮膚纖維母細胞MMP-1蛋白質的表現 42
第十節 探討Shikonin誘發人類皮膚纖維母細胞(Skin fibroblasts)磷酸化Smad3蛋白質的表現 43
第十一節 探討Shikonin誘發人類皮膚纖維母細胞TGF-β蛋白質的表現 44
第四章 討論 46
第一節 活體動物創傷癒合試驗 46
第二節 離體細胞創傷癒合試驗 46
第五章 結論 55
參考文獻 75



圖表目錄
Figure.1-1 Human skin diagram 57
Figure.1-2 The sequence of events during normal wound healing 58
Figure.1-3 Shikonin之化學結構 59
Figure.1-4 The TGF-β/SMAD signaling pathway 60
Figure.3-1 In vivo wound healing assay 61
Figure.3-2 Effects of Shikonin on cell viability after treated for 24hr 62
Figure.3-3 The morphology of human skin fibroblasts 63
Figure.3-4 Shikonin induce ERK1/2 phosphorylation in skin fibroblast cells 64
Figure.3-5 Effects of Shikonin on cell viability after treated for 24hrs and 48hrs 65
Figure.3-6 Shikonin induces cell migration and wound healing in skin fibroblast cells 66
Figure.3-7 Shikonin induce ERK1/2 phosphorylation in skin fibroblast cells 67
Figure.3-8 Shikonin induce expression of PAI-1 in skin fibroblast cells 68
Figure.3-9 Shikonin induce procollagen type I in skin fibroblast cells 69
Figure.3-10 Shikonin induce expression of procollagen type I in skin fibroblast cells 70
Figure.3-11 Shikonin induce expression of MMP-1 in skin fibroblast cells 71
Figure.3-12 Shikonin induce expression of Smad3 phosphorylation in skin fibroblast cells 72
Figure.3-13 Shikonin induce expression of TGFβ in skin fibroblast cells 73
Figure.4-1 The possible pathway of shikonin on human skin fibroblast. 74
參考文獻 Arany, P.R., Flanders, K.C., DeGraff, W., Cook, J., Mitchell, J.B. and
Roberts, A.B. (2007)Absence of Smad3 confers radioprotection
through modulation of ERK-MAPK in primary dermal fibroblasts.
J Dermatol Sci 48, 35-42.
Ashcroft, G.S. and Roberts, A.B. (2000) Loss of Smad3 modulates wound
healing. Cytokine Growth Factor Rev 11, 125-31.
Ashcroft, G.S., Yang, X., Glick, A.B.,Weinstein, M., Letterio, J.L., Mizel,
D.E., Anzano, M., Greenwell-Wild, T.,Wahl, S.M., Deng, C. and
Roberts, A.B. (1999) Mice lacking Smad3 show accelerated wound
healing and an impaired local inflammatory response. Nat Cell
Biol 1, 260-6.
Barford, D., Jia, Z. and Tonks, N.K. (1995) Protein tyrosine phosphatases
take off. Nat Struct Biol 2, 1043-53.
Canty, E.G. and Kadler, K.E. (2005) Procollagen trafficking, processing
and fibrillogenesis. J Cell Sci 118, 1341-53.
Chan, J.C., Duszczyszyn, D.A., Castellino, F.J. and Ploplis, V.A. (2001)
Accelerated skin wound healing in plasminogen activator
inhibitor-1-deficient mice. Am J Pathol 159, 1681-8.
Chen, S.J., Yuan,W., Lo, S., Trojanowska, M. and Varga, J. (2000)
Interaction of smad3 with a proximal smad-binding element of the
human alpha2(I) procollagen gene promoter required for
transcriptional activation by TGF-beta. J Cell Physiol 183, 381-92.
Chen, S.J., Yuan,W., Mori, Y., Levenson, A., Trojanowska, M. and Varga,
J. (1999) Stimulation of type I collagen transcription in human skin
fibroblasts by TGF-beta: involvement of Smad 3. J Invest Dermatol
112, 49-57.
76
Chen, X., Yang, L., Oppenheim, J.J. and Howard, M.Z. (2002) Cellular
pharmacology studies of shikonin derivatives. Phytother Res 16,
199-209.
Cheng, Y.W., Chang, C.Y., Lin, K.L., Hu, C.M., Lin, C.H. and Kang, J.J.
(2008) Shikonin derivatives inhibited LPS-induced NOS in RAW
264.7 cells via downregulation of MAPK/NF-kappaB signaling. J
Ethnopharmacol 120, 264-71.
Cho, S., Kim, H.H., Lee, M.J., Lee, S., Park, C.S., Nam, S.J., Han, J.J.,
Kim, J.W. and Chung, J.H. (2008) Phosphatidylserine prevents
UV-induced decrease of type I procollagen and increase of MMP-1
in dermal fibroblasts and human skin in vivo. J Lipid Res 49,
1235-45.
Cutroneo, K.R. (2003) How is Type I procollagen synthesis regulated at
the gene level during tissue fibrosis. J Cell Biochem 90, 1-5.
Das, F., Ghosh-Choudhury, N., Venkatesan, B., Li, X., Mahimainathan, L.
and Choudhury, G.G. (2008) Akt kinase targets association of CBP
with SMAD 3 to regulate TGFbeta-induced expression of
plasminogen activator inhibitor-1. J Cell Physiol 214, 513-27.
Derynck, R. and Zhang, Y.E. (2003) Smad-dependent and
Smad-independent pathways in TGF-beta family signalling. Nature
425, 577-84.
Dieckgraefe, B.K.,Weems, D.M., Santoro, S.A. and Alpers, D.H. (1997)
ERK and p38 MAP kinase pathways are mediators of intestinal
epithelial wound-induced signal transduction. Biochem Biophys
Res Commun 233, 389-94.
Diegelmann, R.F. and Evans, M.C. (2004)Wound healing: an overview
of acute, fibrotic and delayed healing. Front Biosci 9, 283-9.
Fahey, T.J., 3rd, Sadaty, A., Jones,W.G., 2nd, Barber, A., Smoller, B. and
Shires, G.T. (1991) Diabetes impairs the late inflammatory
response to wound healing. J Surg Res 50, 308-13.
77
Faler, B.J., Macsata, R.A., Plummer, D., Mishra, L. and Sidawy, A.N.
(2006) Transforming growth factor-beta and wound healing.
Perspect Vasc Surg Endovasc Ther 18, 55-62.
Flanders, K.C. (2004) Smad3 as a mediator of the fibrotic response. Int J
Exp Pathol 85, 47-64.
Fujita, H., Omori, S., Ishikura, K., Hida, M. and Awazu, M. (2004) ERK
and p38 mediate high-glucose-induced hypertrophy and TGF-beta
expression in renal tubular cells. Am J Physiol Renal Physiol 286,
F120-6.
Fujita, N., Sakaguchi, I., Kobayashi, H., Ikeda, N., Kato, Y., Minamino,
M. and Ishii, M. (2003) An extract of the root of Lithospermun
erythrorhison accelerates wound healing in diabetic mice. Biol
Pharm Bull 26, 329-35.
Furukawa, F., Matsuzaki, K., Mori, S., Tahashi, Y., Yoshida, K., Sugano,
Y., Yamagata, H., Matsushita, M., Seki, T., Inagaki, Y., Nishizawa,
M., Fujisawa, J. and Inoue, K. (2003) p38 MAPK mediates
fibrogenic signal through Smad3 phosphorylation in rat
myofibroblasts. Hepatology 38, 879-89.
Gao, D., Kakuma, M., Oka, S., Sugino, K. and Sakurai, H. (2000)
Reaction of beta-alkannin (shikonin) with reactive oxygen species:
detection of beta-alkannin free radicals. Bioorg Med Chem 8,
2561-9.
Glading, A., Chang, P., Lauffenburger, D.A. andWells, A. (2000)
Epidermal growth factor receptor activation of calpain is required
for fibroblast motility and occurs via an ERK/MAP kinase
signaling pathway. J Biol Chem 275, 2390-8.
Guo, B., Inoki, K., Isono, M., Mori, H., Kanasaki, K., Sugimoto, T.,
Akiba, S., Sato, T., Yang, B., Kikkawa, R., Kashiwagi, A., Haneda,
M. and Koya, D. (2005) MAPK/AP-1-dependent regulation of
PAI-1 gene expression by TGF-beta in rat mesangial cells. Kidney
Int 68, 972-84.
78
Hayashida, T., Decaestecker, M. and Schnaper, H.W. (2003) Cross-talk
between ERK MAP kinase and Smad signaling pathways enhances
TGF-beta-dependent responses in human mesangial cells. Faseb J
17, 1576-8.
Heldin, C.H., Miyazono, K. and ten Dijke, P. (1997) TGF-beta signalling
from cell membrane to nucleus through SMAD proteins. Nature
390, 465-71.
Hertig, A. and Rondeau, E. (2004) Plasminogen activator inhibitor type 1:
the two faces of the same coin. Curr Opin Nephrol Hypertens 13,
39-44.
Hong, H.J., Jin, S.E., Park, J.S., Ahn,W.S. and Kim, C.K. (2008)
Accelerated wound healing by smad3 antisense
oligonucleotides-impregnated chitosan/alginate polyelectrolyte
complex. Biomaterials 29, 4831-7.
Huang, J.S.,Wang, Y.H., Ling, T.Y., Chuang, S.S., Johnson, F.E. and
Huang, S.S. (2002) Synthetic TGF-beta antagonist accelerates
wound healing and reduces scarring. Faseb J 16, 1269-70.
Ishida, T. and Sakaguchi, I. (2007) Protection of human keratinocytes
from UVB-induced inflammation using root extract of
Lithospermum erythrorhizon. Biol Pharm Bull 30, 928-34.
Itoh, S., Itoh, F., Goumans, M.J. and Ten Dijke, P. (2000) Signaling of
transforming growth factor-beta family members through Smad
proteins. Eur J Biochem 267, 6954-67.
Johnson, G.L. and Lapadat, R. (2002) Mitogen-activated protein kinase
pathways mediated by ERK, JNK, and p38 protein kinases.
Science 298, 1911-2.
Kavitha, O. and Thampan, R.V. (2008) Factors influencing collagen
biosynthesis. J Cell Biochem 104, 1150-60.
79
Kim, H.S., Shang, T., Chen, Z., Pflugfelder, S.C. and Li, D.Q. (2004)
TGF-beta1 stimulates production of gelatinase (MMP-9),
collagenases (MMP-1, -13) and stromelysins (MMP-3, -10, -11) by
human corneal epithelial cells. Exp Eye Res 79, 263-74.
Kim, M.S., Kim, Y.K., Cho, K.H. and Chung, J.H. (2006) Regulation of
type I procollagen and MMP-1 expression after single or repeated
exposure to infrared radiation in human skin. Mech Ageing Dev
127, 875-82.
Kutz, S.M., Hordines, J., McKeown-Longo, P.J. and Higgins, P.J. (2001)
TGF-beta1-induced PAI-1 gene expression requires MEK activity
and cell-to-substrate adhesion. J Cell Sci 114, 3905-14.
Kyriakis, J.M., Banerjee, P., Nikolakaki, E., Dai, T., Rubie, E.A., Ahmad,
M.F., Avruch, J. andWoodgett, J.R. (1994) The stress-activated
protein kinase subfamily of c-Jun kinases. Nature 369, 156-60.
Leask, A. and Abraham, D.J. (2004) TGF-beta signaling and the fibrotic
response. Faseb J 18, 816-27.
Lee, H.J., Lee, H.J., Magesh, V., Nam, D., Lee, E.O., Ahn, K.S., Jung,
M.H., Ahn, K.S., Kim, D.K., Kim, J.Y. and Kim, S.H. (2008)
Shikonin, Acetylshikonin, and Isobutyroylshikonin Inhibit
VEGF-induced Angiogenesis and Suppress Tumor Growth in
Lewis Lung Carcinoma-bearing Mice. Yakugaku Zasshi 128,
1681-8.
Liu, X., Sun, Y., Constantinescu, S.N., Karam, E., Weinberg, R.A. and
Lodish, H.F. (1997) Transforming growth factor beta-induced
phosphorylation of Smad3 is required for growth inhibition and
transcriptional induction in epithelial cells. Proc Natl Acad Sci U S
A 94, 10669-74.
Lobmann, R., Ambrosch, A., Schultz, G.,Waldmann, K., Schiweck, S.
and Lehnert, H. (2002) Expression of matrix-metalloproteinases
and their inhibitors in the wounds of diabetic and non-diabetic
patients. Diabetologia 45, 1011-6.
80
Loots, M.A., Lamme, E.N., Zeegelaar, J., Mekkes, J.R., Bos, J.D. and
Middelkoop, E. (1998) Differences in cellular infiltrate and
extracellular matrix of chronic diabetic and venous ulcers versus
acute wounds. J Invest Dermatol 111, 850-7.
Lund, L.R., Green, K.A., Stoop, A.A., Ploug, M., Almholt, K., Lilla, J.,
Nielsen, B.S., Christensen, I.J., Craik, C.S.,Werb, Z., Dano, K. and
Romer, J. (2006) Plasminogen activation independent of uPA and
tPAmaintains wound healing in gene-deficient mice. Embo J 25,
2686-97.
Lynch, S.E., Colvin, R.B. and Antoniades, H.N. (1989) Growth factors in
wound healing. Single and synergistic effects on partial thickness
porcine skin wounds. J Clin Invest 84, 640-6.
Mani, H., Sidhu, G.S., Singh, A.K., Gaddipati, J., Banaudha, K.K., Raj, K.
and Maheshwari, R.K. (2004) Enhancement of wound healing by
shikonin analogue 93/637 in normal and impaired healing. Skin
Pharmacol Physiol 17, 49-56.
Martin, P. (1997)Wound healing--aiming for perfect skin regeneration.
Science 276, 75-81.
Massague, J. and Chen, Y.G. (2000) Controlling TGF-beta signaling.
Genes Dev 14, 627-44.
Massague, J. andWotton, D. (2000) Transcriptional control by the
TGF-beta/Smad signaling system. Embo J 19, 1745-54.
Matsubayashi, Y., Ebisuya, M., Honjoh, S. and Nishida, E. (2004) ERK
activation propagates in epithelial cell sheets and regulates their
migration during wound healing. Curr Biol 14, 731-5.
McAnulty, R.J., Chambers, R.C. and Laurent, G.J. (1995) Regulation of
fibroblast procollagen production. Transforming growth factor-beta
1 induces prostaglandin E2 but not procollagen synthesis via a
pertussis toxin-sensitive G-protein. Biochem J 307 ( Pt 1), 63-8.
81
Midwood, K.S.,Williams, L.V. and Schwarzbauer, J.E. (2004) Tissue
repair and the dynamics of the extracellular matrix. Int J Biochem
Cell Biol 36, 1031-7.
Mimuro, J. (1991) [Type 1 plasminogen activator inhibitor: its role in
biological reactions]. Rinsho Ketsueki 32, 487-9.
Miyazono, K., ten Dijke, P. and Heldin, C.H. (2000) TGF-beta signaling
by Smad proteins. Adv Immunol 75, 115-57.
Montesano, R. and Orci, L. (1988) Transforming growth factor beta
stimulates collagen-matrix contraction by fibroblasts: implications
for wound healing. Proc Natl Acad Sci U S A 85, 4894-7.
Mori, Y., Chen, S.J. and Varga, J. (2000) Modulation of endogenous
Smad expression in normal skin fibroblasts by transforming growth
factor-beta. Exp Cell Res 258, 374-83.
Mulder, K.M. (2000) Role of Ras and Mapks in TGFbeta signaling.
Cytokine Growth Factor Rev 11, 23-35.
Nikkari, S.T., Geary, R.L., Hatsukami, T., Ferguson, M., Forough, R.,
Alpers, C.E. and Clowes, A.W. (1996) Expression of collagen,
interstitial collagenase, and tissue inhibitor of metalloproteinases-1
in restenosis after carotid endarterectomy. Am J Pathol 148,
777-83.
O'Kane, S. and Ferguson, M.W. (1997) Transforming growth factor beta s
and wound healing. Int J Biochem Cell Biol 29, 63-78.
Ozaki, Y., Ohno, A., Saito, Y. and Satake, M. (1994) Accelerative effect
of shikonin, alkannin and acetylshikonin on the proliferation of
granulation tissue in rats. Biol Pharm Bull 17, 1075-7.
82
Ozgen, U., Ikbal, M., Hacimuftuoglu, A., Houghton, P.J., Gocer, F.,
Dogan, H. and Coskun, M. (2006) Fibroblast growth stimulation by
extracts and compounds of Onosma argentatum roots. J
Ethnopharmacol 104, 100-3.
Parks,W.C. (1999) Matrix metalloproteinases in repair.Wound Repair
Regen 7, 423-32.
Providence, K.M. and Higgins, P.J. (2004) PAI-1 expression is required
for epithelial cell migration in two distinct phases of in vitro wound
repair. J Cell Physiol 200, 297-308.
Quan, T., He, T., Kang, S., Voorhees, J.J. and Fisher, G.J. (2002)
Ultraviolet irradiation alters transforming growth factor beta/smad
pathway in human skin in vivo. J Invest Dermatol 119, 499-506.
Raghow, R., Postlethwaite, A.E., Keski-Oja, J., Moses, H.L. and Kang,
A.H. (1987) Transforming growth factor-beta increases steady state
levels of type I procollagen and fibronectin messenger RNAs
posttranscriptionally in cultured human dermal fibroblasts. J Clin
Invest 79, 1285-8.
Ravanti, L. and Kahari, V.M. (2000) Matrix metalloproteinases in wound
repair (review). Int J Mol Med 6, 391-407.
Ravanti, L., Toriseva, M., Penttinen, R., Crombleholme, T., Foschi, M.,
Han, J. and Kahari, V.M. (2001) Expression of human
collagenase-3 (MMP-13) by fetal skin fibroblasts is induced by
transforming growth factor beta via p38 mitogen-activated protein
kinase. Faseb J 15, 1098-100.
Sakaguchi, I., Tsujimura, M., Ikeda, N., Minamino, M., Kato, Y.,Watabe,
K., Yano, I. and Kaneda, K. (2001) Granulomatous tissue
formation of shikon and shikonin by air pouch method. Biol Pharm
Bull 24, 650-5.
83
Schiller, M., Javelaud, D. and Mauviel, A. (2004) TGF-beta-induced
SMAD signaling and gene regulation: consequences for
extracellular matrix remodeling and wound healing. J Dermatol Sci
35, 83-92.
Sharma, G.D., He, J. and Bazan, H.E. (2003) p38 and ERK1/2 coordinate
cellular migration and proliferation in epithelial wound healing:
evidence of cross-talk activation between MAP kinase cascades. J
Biol Chem 278, 21989-97.
Singer, A.J. and Clark, R.A. (1999) Cutaneous wound healing. N Engl J
Med 341, 738-46.
Singh, F., Gao, D., Lebwohl, M.G. andWei, H. (2003) Shikonin
modulates cell proliferation by inhibiting epidermal growth factor
receptor signaling in human epidermoid carcinoma cells. Cancer
Lett 200, 115-21.
Soo, C., Shaw, W.W., Zhang, X., Longaker, M.T., Howard, E.W. and Ting,
K. (2000) Differential expression of matrix metalloproteinases and
their tissue-derived inhibitors in cutaneous wound repair. Plast
Reconstr Surg 105, 638-47.
Southern, S.J. and Browning, F.S. (1995) A skin model for wound closure
tuition. Injury 26, 213-4.
Takehara, K. (2000) Growth regulation of skin fibroblasts. J Dermatol Sci
24 Suppl 1, S70-7.
Ten Dijke, P., Goumans, M.J., Itoh, F. and Itoh, S. (2002) Regulation of
cell proliferation by Smad proteins. J Cell Physiol 191, 1-16.
Vassalli, J.D., Sappino, A.P. and Belin, D. (1991) The plasminogen
activator/plasmin system. J Clin Invest 88, 1067-72.
84
Vayalil, P.K., Iles, K.E., Choi, J., Yi, A.K., Postlethwait, E.M. and Liu,
R.M. (2007) Glutathione suppresses TGF-beta-induced PAI-1
expression by inhibiting p38 and JNK MAPK and the binding of
AP-1, SP-1, and Smad to the PAI-1 promoter. Am J Physiol Lung
Cell Mol Physiol 293, L1281-92.
Wang, J.F., Olson, M.E., Reno, C.R., Kulyk,W., Wright, J.B. and Hart,
D.A. (2000) Molecular and cell biology of skin wound healing in a
pig model. Connect Tissue Res 41, 195-211.
Wayne, J., Sielski, J., Rizvi, A., Georges, K. and Hutter, D. (2006) ERK
regulation upon contact inhibition in fibroblasts. Mol Cell Biochem
286, 181-9.
Werner, S. and Grose, R. (2003) Regulation of wound healing by growth
factors and cytokines. Physiol Rev 83, 835-70.
Wrana, J.L. and Attisano, L. (2000) The Smad pathway. Cytokine Growth
Factor Rev 11, 5-13.
Wu, J.J. and Bennett, A.M. (2005) Essential role for mitogen-activated
protein (MAP) kinase phosphatase-1 in stress-responsive MAP
kinase and cell survival signaling. J Biol Chem 280, 16461-6.
Yang, C., Patel, K., Harding, P., Sorokin, A. and Glass,W.F., 2nd (2007)
Regulation of TGF-beta1/MAPK-mediated PAI-1 gene expression
by the actin cytoskeleton in human mesangial cells. Exp Cell Res
313, 1240-50.
Yang, F., Chen, Y., Duan,W., Zhang, C., Zhu, H. and Ding, J. (2006)
SH-7, a new synthesized shikonin derivative, exerting its potent
antitumor activities as a topoisomerase inhibitor. Int J Cancer 119,
1184-93.
Yang, L., Cranson, D. and Trinkaus-Randall, V. (2004) Cellular injury
induces activation of MAPK via P2Y receptors. J Cell Biochem 91,
938-50.
85
Yuan,W. and Varga, J. (2001) Transforming growth factor-beta repression
of matrix metalloproteinase-1 in dermal fibroblasts involves Smad3.
J Biol Chem 276, 38502-10.
Yue, J. and Mulder, K.M. (2000) Requirement of Ras/MAPK pathway
activation by transforming growth factor beta for transforming
growth factor beta 1 production in a Smad-dependent pathway. J
Biol Chem 275, 30765-73.
Zhang, Y. and Derynck, R. (1999) Regulation of Smad signalling by
protein associations and signalling crosstalk. Trends Cell Biol 9,
274-9.
Zhou, A., Huntington, J.A., Pannu, N.S., Carrell, R.W. and Read, R.J.
(2003) How vitronectin binds PAI-1 to modulate fibrinolysis and
cell migration. Nat Struct Biol 10, 541-4.

------------------------------------------------------------------------ 第 8 筆 ---------------------------------------------------------------------
系統識別號 U0007-1208200915104300
論文名稱(中文) 閉鞘薑抗乳癌之活性成分研究
論文名稱(英文) The Antitumor Constituents of Costus speciosus in Human Breast Adenocarcinoma
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 97
學期 2
出版年 98
研究生(中文) 胡雪吟
學號 M303096003
學位類別 碩士
語文別 中文
口試日期 2009-07-13
論文頁數 95頁
口試委員 指導教授-王靜瓊
委員-李哲夫
委員-李連滋
委員-許秀蘊
委員-鄭幼文
關鍵字(中) 閉鞘薑
乳癌
癌細胞毒性
P-388D1擔癌鼠
關鍵字(英) Costus speciosus.
breast adenocarcinoma
antitumor effect
P-388D1 bearing mice
學科別分類
中文摘要   乳癌為台灣女性癌症發生率第一位,如何抑制癌細胞生長是重要的課題。本研究取二十種台灣植物之50%甲醇迴流萃物,以兩株乳癌細胞(MDA-MB-231和 MCF-7細胞株)作為靶細胞,進行體外細胞毒性檢測,並配合管柱分離,預期得到具有抑制癌細胞生長之起始天然物,結果顯示薑科植物閉鞘薑(Costus speciosus (Koenig) Smith)之根莖部抑制癌細胞生長作用較為明顯,繼而以水和乙酸乙酯作分部分析,經由Diaion HP-20管柱,以不同比例的水和甲醇沖提,分離得到5個劃分部,其中D80M和D100M具有較強的細胞毒性,且D100M具有意義的延長P-388D1之CDF1擔癌鼠存活時間。因此再利用VersaPak® Flash、Silica gel等不同分離管柱,純化D80M和D100M得到6個主要成分,分別為(25R)-neospirost-4en-3-one (1)、25-hydroxyspirost-4-en-3-one (2)、β-D-Glucopyranoside, (3β, 25R)-17-hydroxyspirost-en-3-yl 2-Ο-(6-deoxy-α-L-mannopyranosyl) (3)、Prosaponin A of dioscin (4)、dioscin (5)和gracillin (6)。其中又以Prosaponin A of dioscin、dioscin、gracillin對人類乳癌細胞具有細胞毒性,IC50分別對MDA-MB-231細胞株分別為4.88、3.43、5.37 μM,對MCF-7細胞株分別為4.6、3.39、3.36 μM,且會使乳癌細胞PARP降解,引起凋亡。另外,dioscin亦能有意義的延長P-388D1之CDF1擔癌鼠存活時間。綜合上述,閉鞘薑之薯蕷皂苷類化合物於體內、外實驗皆有活性,能引起乳癌細胞凋亡,而dioscin可作為閉鞘薑抗癌活性之指標成分或做為開發抗癌藥物的起始結構,為一個值得發展的抗癌經濟作物。
論文目次 考試委員名錄
誌謝
目錄………………………………………………………………………I
圖目錄…………………………………………………………………III
表目錄…………………………………………………………………V
縮寫表…………………………………………………………………VII
中文摘要…………………………………………………………………1
英文摘要…………………………………………………………………3
緒論………………………………………………………………………5
實驗目的…………………………………………………………………26
實驗部份…………………………………………………………………27
1.材料……………………………………………………………………27
2.儀器……………………………………………………………………30
3.實驗方法………………………………………………………………31
3-1. 二十種台灣原生種植物成分萃取………………………………31
3-2. 細胞毒性篩選方法………………………………………………31
3-3. 閉鞘薑根莖部成分萃取、分離與純化…………………………34
3-4. 閉鞘薑成分之結構鑑定…………………………………………38
3-5. P-388D1誘導擔癌鼠之體內抗癌模式…………………………38
3-6. 西方點墨法分析相關蛋白的表現………………………………39
3-7.閉鞘薑萃取物指紋圖譜之分析…………………………………41
4.實驗結果……………………………………………………………42
4-1.二十種植物對乳癌細胞株之抑制活性…………………………42
4-2.閉鞘薑各部位之細胞毒性試驗…………………………………43
4-3.閉鞘薑根莖部之結構解析………………………………………46
4-4.閉鞘薑萃取物指紋圖譜之分析…………………………………51
4-5.閉鞘薑誘導之細胞凋亡…………………………………………62
4-6.閉鞘薑對P-388D1誘導擔癌鼠之體內抗癌活性試驗……………68
討論……………………………………………………………………74
參考文獻………………………………………………………………78
附錄……………………………………………………………………84
參考文獻 1. 行政院衛生署, 96年公告. (2007).
2. 黃俊升, 成佳憲, 王正旭, & 俞志誠, 希望之路面對乳癌. 中華癌症希望協會 (2006).
3. 陳訓徹, 先化學治療,再開刀-談乳癌先期性化療的進展. 癌症新探 30 (2009).
4. 李明陽, 乳癌化學治療的最新發展. 癌症新探 34 (2009).
5. 廖繼鼎. 臨床腫瘤學 (2003).
6. Lehne, R.A., Linda, A.M., Leanna, J.C., & Diane, B.H., Pharmacology for nursing care. 4th edition. Saunders. (2001).
7. Chisholm, M.A. et al., pharmacotherapy principles & practice. 1310-1322 (2008).
8. Koda-Kimble, M.A. & Young, L.Y., Handbook of Applied Therapeutics, 合記. (2002).
9. 萬騁 et al., 抗腫瘤血管生成藥bevacizumab對VEGF促人肝癌細胞株HepG2增殖的阻斷作用 第二軍醫大學學報 29 (9) (2008)
10. 邵波, 癌癥的中醫理論溯源. 環球中醫藥 01 (2008).
11. 曹榮穎, 中醫調理乳癌. (2007).
12. 陳俊明. 台灣新生報 (2007).
13. 王曉 et al., 中醫在婦科惡性腫瘤治療中的作用和地位. 台灣中醫科學雜誌 3 (1) (2008).
14. Cragg, G.M., Paclitaxel (Taxol): a success story with valuable lessons for natural product drug discovery and development. . Med Res Rev. 18 (5), 315-331 (1998).
15. 楊遠波, 劉和義, & 林讚標, 台灣維管束植物簡誌. 206 (2001).
16. 張憲昌. 台灣藥用植物圖鑑, 87 (2007).
17. 台北植物園資訊網 http://tpbg.tfri.gov.tw/plants/plants_info.aspx?rid=230.
18. Tewari, P.V., Chaturvedi, C., Pandey, V.B. , Antifertility activity of Costus speciosus Sm. . Indian Journal of Pharmacy 35 (4), 114-115 (1973).
19. Sharma, B.B., Varshney, M.D., Gupta, D.N., & Prakash, A.O., Antifertility screening of plants. Part I. Effect of ten indigenous plants on early pregnancy in albino rats. International Journal of Crude Drug Research 21 (4), 183-187 (1983).
20. Bora, U., Sahu, A., Saikia, A.P., Ryakala, V.K., Goswami, P. , Medicinal plants used by the people of Northeast India for curing malaria. Phytotherapy Research 21 (8), 800-804 (2007).
21. 陳昌祥 & 銀慧新, 閉鞘薑根中的甾體皂苷. 天然產物研究與開發 7 (4), 18-22 (1995).
22. 李忠瓊 et al., 閉鞘薑屬植物化學成分及藥理研究概況. 中藥材 24 (2), 148-150 (2001).
23. Ratnayake Bandara, B.M., Hewage, C.M., Karunaratne, V., & Adikaram, N.K.B., Methyl ester of para-coumaric acid: Antifungal principle of the rhizome of Costus speciosus. Planta Medica 54 (5), 477-478 (1988).
24. Mosihuzzaman, M. et al., Hypoglycemic effects of three plants from eastern Himalayan belt. Diabetes Research 26 (3), 127-138 (1994).
25. Bavarva, J.H. & Narasimhacharya, A.V.R.L., Antihyperglycemic and hypolipidemic effects of Costus speciosus in alloxan induced diabetic rats. Phytotherapy Research 22 (5), 620-626 (2008).
26. Eliza, J., Daisy, P., Ignacimuthu, S., & Duraipandiyan, V., Normo-glycemic and hypolipidemic effect of costunolide isolated from Costus speciosus (Koen ex. Retz.)Sm. in streptozotocin-induced diabetic rats. Chem Biol Interact (2008).
27. Vijayalakshmi, M.A. & Sarada, N.C., Screening of Costus speciosus extracts for antioxidant activity. Fitoterapia 79 (3), 197-198 (2008).
28. Singh, S., Sanyal, A.K., Bhattacharya, S.K., & Pandey, V.B., Oestrogenic activity of saponins from Costus speciosus (Koen) Sm. The Indian journal of medical research 60 (2), 287-290 (1972).
29. HUO, R. et al., Diosgenin induces apoptosis in HeLa cells via activation of caspase pathway. Acta Pharmacologica Sinica 25 (8), 1077-1082 (2004).
30. Liu, M.-J., Wang, Z., Ju, Y., Wong, R.N.-S., & Wu, Q.-Y., Diosgenin induces cell cycle arrest and apoptosis in human leukemia K562 cells with the disruption of Ca2+ homeostasis. Cancer Chemother Pharmacol 55, 79-90 (2005).
31. Wang, Y., Che, C.-M., Chiu, J.-F., & He, Q.-Y., Dioscin (Saponin)-Induced Generation of Reactive Oxygen Species through Mitochondria Dysfunction: A Proteomic-Based Study. Journal of Proteome Research 6, 4703-4710 (2007).
32. Jing, C., Mingjie, L., Zhao, W., & Yong, J., Apoptosis Induced by Dioscin in Hela Cells. Biol. Pharm. Bull. 25 (2), 193-196 (2002).
33. Hu, K., Dong, A., Yao, X., Kobayashi, H., & Iwasaki, S., Antineoplastic agents; I. Three spirostanol glycosides from rhizomes of Dioscorea collettii var. hypoglauca. Planta Medica 62 (6), 573-575 (1996).
34. Gupta, M.M., Farooqui, S.U., Lal, R.N., Distribution and variation of diosgenin in different parts of Costus speciosus. . Journal of Natural Products 44 (4), 486-489 (1981).
35. 魏金婷 & 劉文, 植物藥活性成分β-谷甾醇研究概況. 莆田學院學報 14 (2) (2007).
36. Chiang, H.C., Tseng, T.H., Wang, C.J., Chen, C.F., & Kan, W.S., Experimental antitumor agents from Solanum indicum L. Anticancer Research 11 (5), 1911-1917 (1991).
37. Chiang, H.C., Wang, J.J., & Wu, R.T., Immunomodulating effects of the hydrolysis products of formosanin C and β-Ecdysone from Paris formosana Hayata. Anticancer Research 12 (5), 1475-1478 (1992).
38. Hu, K., Dong, A., Yao, X., Kobayashi, H., & Iwasaki, S., Antineoplastic agents; II. Four furostanol glycosides from rhizomes of Dioscorea collettii var. hypoglauca. Planta Medica 63 (2), 161-165 (1997).
39. Hu, K. & Yao, X., The cytotoxicity of methyl protoneogracillin (NSC-698793) and gracillin (NSC-698787), two steroidal saponins from the rhizomes of Dioscorea collettii var. hypoglauca, against human cancer cells in vitro. Phytotherapy Research 17 (6), 620-626 (2003).
40. Julin, L. & Van, L., Apoptosis, genomic integrity, and cancer. . Jones and Barlett publishers 61-65, 133-140, 175-179 (2006).
41. Pistilli, E.E., Jackson, J.R., & Alway, S.E., Death receptor-associated pro-apoptotic signaling in aged skeletal muscle. . Apoptosis 11, 2115-2126 (2006).
42. Yang, P. et al., Caspase-8-mediated apoptosis in human RPE cells. . Invest Ophthalmol Vis Sci 48, 3341-3349 (2007).
43. Knudson, C.M.a.B., N.M. , Mitochondria potential, bax "activation," and programmed cell death. . Methods Mol Biol 414, 95-108 (2008).
44. Cregan, S.P., Dawson, V.L. and Slack, R.S. , Role of AIF in caspase-dependent and caspase-independent cell death. . Oncogene 23, 2785-2796 (2004).
45. Krantic, S., Mechawar, N., Reix, S. and Quirion, R. , Apoptosis-inducing factor: a matter of neuron life and death. . Prog. Neurobiol 81 179-196 (2007).
46. Wang, X., The expanding role of mitochondria in apoptosis. . Genes and Development 15, 2922-2933 ( 2001).
47. Niles, A.L., Moravec, R.A. and Riss, T.L. , Multiplex caspase activity and cytotoxicity assays. . Methods Mol Biol 414, 151-162 (2008).
48. Ha, H., Park, J., Kim, Y.S. and Endou, H. , Oxidative stress and chronic allograft nephropathy. . Yonsei Med J 45, 1049-1052 (2004).
49. Fleury, C., Mignotte, B. and Vayssiere, J.L. , Mitochondrial reactive oxygen species in cell death signaling. . Biochimie 84, 131-141 ( 2002).
50. Cejas, P., Casado, E., Belda-Iniesta, C., De Castro, J., Espinosa, E., Redondo, A., Sereno, M., Garcia-Cabezas, M.A., Vara, J.A., Dominguez-Caceres, A., Perona, R. and Gonzalez-Baron, M. , Implications of oxidative stress and cell membrane lipid peroxidation in human cancer (Spain). . Cancer Causes Control 15, 707-719 (2004).
51. AGNES GÖRLACH, P.K., and THOMAS KIETZMANN, The Endoplasmic Reticulum: Folding, Calcium Homeostasis, Signaling, and Redox Control. ENDOPLASMIC RETICULUM 8 (9,10), 1391-1418 (2006).
52. Shoei-Sheng Lee, J.-L.Y., Gong-Yin Lin, and K. C. Wang, Microbial Transformation of Dihydrosarsasapogenin with Mycobacterium sp. Journal of Natural Products 61, 275-278 (1998).
53. Hans Achenbach, H.H., Melchior Reiter, Cholestane- and Pregnane-type Glycosides from the Roots of Tribulus Cistoides Phytochemistry 41 (3), 907-917 (1996).
54. Z.-H. Cheng, T.W., B.-Y. Yu, Steroidal glycosides from tubers of Ophiopogon japonicus. Journal of Asian Natural Products Research 8 (6), 555-559 (2006).
55. 馬朋、曹同濤、劉棟、姜濤、王慧林, 薯蕷皂苷類化合物體外抗腫瘤作用的研究. 濱州醫學院學報 31 (5) (2008).
56. Amigo, L., Mendoza, H., Zanlungo, S., Miquel, J. F., Rigotti, A., Gonzalz, S., Nervi, F., Enrichment of canalicular membrane with cholesterol and sphingomylin prevents bile salt-induced hepatic damage. J. Lipid Res. 40, 533-542 (1999).
57. Cayen, M.N.D., D. , Effect of diosgenin on lipid metabolism in rats. . J. Lipid Res. 20 (12), 162-174 (1979).
58. Yamada, T., Hosshino, M., Hayakawa, T., Ohhara, H., Yamada, H., Nakazawa, T., Inagaki, T., Iida, M., Miyaji, M., Hirata, A., Takeuchi, T. , Dietary diogenin attenuates subacute intestinal inflammation associatted with indomethacin in rats. . Am. J. Physiol. 213, 355-364 (1997).
59. Chao-Chin Hu, J.-T.L., Shih-Chuan Liu, Deng-Jye Yang, A Spirostanol Glycoside from Wild Yam (Dioscorea villosa) Extract and Its Cytostatic Activity on Three Cancer Cells. Journal of Food and Drug Analysis 15 (3), 310-315 (2007).
60. Kawasaki, T., Yamauchi, T., Structures of prosapogenin-B and -A of dioscin and cooccurrence of B with dioscin in the rhizoma of Dioscorea tokoro Makino. Chemical and Pharmaceutical Bulletin 16 (6), 1070-1075 (1968).
61. Singh, S.B., Thakur, R.S. , Saponins From the Seeds of Costus speciosus. Journal of Natural Products 45 (6), 667-671 (1982).
62. Inoue, K., Kobayashi, S., Noguchi, H., Sankawa, U., Ebizuka, Y. , Spirostanol and furostanol glycosides of Costus speciosus (Koenig.) SM. . Natural Medicines 49 (3), 336-339 (1995).

------------------------------------------------------------------------ 第 9 筆 ---------------------------------------------------------------------
系統識別號 U0007-1307200909265400
論文名稱(中文) 5,6,7-trimethoxyindoles 和 5,6,7-trimethoxy-2-oxoindoles 抗癌化合物之合成和結構活性關係研究
論文名稱(英文) Synthesis and Structure Activity Relationship of 5,6,7-trimethoxyindoles and 5,6,7-trimethoxy-2-oxoindoles as anticancer agents
校院名稱 臺北醫學大學
系所名稱(中) 藥學研究所
系所名稱(英) Graduate Institute of Pharmacy
學年度 97
學期 2
出版年 98
研究生(中文) 張齡尹
學號 M301096008
學位類別 碩士
語文別 中文
口試日期 2009-06-26
論文頁數 132頁
口試委員 委員-陳繼明
委員-李慶國
指導教授-劉景平
關鍵字(中)
關鍵字(英) Cancer
學科別分類
中文摘要 目前具有發展性的抗癌藥物是antimitotic agents;例如 taxanes 和 vinca alkaloids。但是以上化合物具有生理毒性、複雜合成、抗藥性以及繁複的分離方式問題,所以科學家想發展新的 antimitotic agents 來克服以上問題。根據文獻,Combretastatin A-4 [ cis-1-(3,4,5-trimethoxyphenyl)-2-(3-hydroxy-4-methoxyphenyl) ethane; CA4] 是目前最有發展性的物質。它主要是結合至colchicine binding domain,它主要作用是可以快速使微管束快速depolymerization,因此改變血管內皮細胞形狀,封閉已存在的腫瘤血管,最後促使癌細胞缺乏氧和養份而死亡。
Combretastatin A-4P(CA4P, ZYBRESTATTM)是目前發展快速的VDA。因此很多藥化學家模仿它的形狀,創造許多的新的相似物。因此,在設計活性分子的結構,同樣基於 bioisosterism 的概念,將 A ring 改變,來進行研發具有活性的分子。我們以五環的方式,在空間上形成與 CA4 相似的結構,形成5,6,7-trimethoxyindoles和5,6,7-trimethoxy-2-oxoindoles系列,以期待具有生物活性。針對這兩個主結構,進行修飾。
在主結構5,6,7-trimethoxyindole上,我們以各種aryl, aroyl, arylthio, benzyl, arylsufone等修飾3位,形成化合物1-8;另外以各種substituted benzyl/heterocyclic group等修飾1位,形成化合物9-15。在主結構5,6,7-trimethoxy-2-oxoindole上,我們將之接上各種substituted benzyl/heterocyclic aldehyde等修飾主結構,形成化合物16-23。以上化合物等來進行人類口腔癌細胞的抑制活性測試(IC50)。
從活性結果從顯示在以5,6,7-trimethoxy-2-oxoindole為主體的化合物,不論接上何種基團活性都大於10 μM。而在5,6,7-trimethoxyindole骨架的3號位置做修飾,活性約在IC50 > 2,5 μM. 其indole環上1號位置以aryl 官能基做修飾,其活性是在這三個系列中,活性較佳的。例如以化合物11和13,其抑制口腔癌細胞生長之為31nM和17nM。

英文摘要 One of currently useful chemotherapy drugs in oncology is represented by antimitotic agents, for example, taxanes and vinca alkaloids. However, the issues of high systemic toxicity, complex syntheses, drug resistance, and isolation procedures have encouraged scientists to develop new antimitotic agents. Recent literatures reported that the antitubulin agents targeting at the colchicine-binding domain, rapidly depolymerize microtubules of vasculatures changing morphology in the endothelial cells of tumor’s vessels to block the blood supply to tumors, can act as vascular-disrupting agents, for example, compound combretastatin A-4P (CA4P) and ZD-6126. The encouraging antivascular and antitumor prolife of CA4P has stimulated pronounced interest in design and synthesis of variety of derivatives or analogues. Here we report our attempt to explore the 5,6,7-trimethoxyindoles or 5,6,7-trimethoxy-2-oxoindoles core coupled with the various aryl, arylthio, arylsulfone, and aroyl groups as anticancer agents. The structure-activity relationship studies of series of 5,6,7-trimethoxyindoles led to discovery of N1-heteroaryl-5,6,7-trimethoxy-indoles as novel potent anticancer agents, for example, compound 11 and 13, showing antiproliferative activity with IC50 values of 11 and 17 nM, respectively.
論文目次 目錄………………………………………………………………Ⅰ
附圖目錄…………………………………………………………Ⅲ
附錄實驗部份目錄…………………..……………………………..Ⅳ
附錄附圖部份目錄……………………………………………………Ⅵ
中文摘要.......................................... 1
英文摘要...........................................2
研究背景........................................... 3
研究目的與設計.................................... 10
結果及討論.......................................... 13
一、5,6,7-trimethoxyindoles…………………………………………….13
1. 化學結構設計…………………………………………………13
2. 合成流程及方式……………………………..………………..13
3. 抗癌細胞活性試驗結果………………………………...17
4. 化學結構與抗癌活性的關係…………………………………18
二、5,6,7- trimethoxy-2-oxoindole………………………………19
1. 化學結構設計……………………………………………………..19
2. 合成流程及方式……………………………………………………19
3. 抗癌細胞活性試驗結果……………………………………………23
4. 化學結構與抗癌活性的關係………………………………………24
生物活性實驗方法.................................... 25
結論……………………………………………………26
實驗儀器........................................... 27
試藥與試劑......................................... 28
參考資料............................................... 31
實驗附份………………………………………………………33
附圖部份……………………………………………71
參考文獻 1.Attard, G.; Greystoke, A.; Kaye, S.; Bono, J. D. Update on tubulin-binding agents. Pathologie Biologie 2006, 54, 72-84.
2.Gaya, A. M.; Rustin, G. J. S. Vascular Disrupting Agents: A New Class of Drug in Cancer Therapy Clinical Oncology 2005, 17, 277-290.
3.Jordan, A.; Hadfield, J. A.; Lawrence, N. J.; McGown, A. T. Tubulin as a Target for Anticancer Drugs: Agents Which Interact with the Mitotic Spindle. Medicinal research reviews 1998, 18, 259-296.
4.Lippert, J. W. Vascular disrupting agents. Biological & Medicinal Chemistry 2007, 15, 605-615.
5.Tozer, G. M.; Kanthou, C.; Baguley, B. C. Distrupting Tumour Blood Vessels. Nature Reviews. Cancer. 2005, 5, 423-435.
6.Liou, J.-P.; Chang, Y.-L.; Kuo, F.-M.; Chang, C.-W.; Tseng, H.-Y.; Wang, C.-C.; Yang, Y.-N.; Chang, J.-Y.; Lee, S.-J.; Hsieh, H.-P. Concise Synthesis and Structure−Activity Relationships of Combretastatin A-4 Analogues, 1-Aroylindoles and 3-Aroylindoles, as Novel Classes of Potent Antitubulin Agents. Journal of Medicinal Chemistry 2004, 47, 4247-4257.
7.Magnus, P.; Gazzard, L.; Hobson, L.; Payne, A. H.; Rainey, T. J.; Westlund, N.; Lynch, V. Synthesis of the Kopsia alkaloids (±)-pauciflorine B, (±)-lahadinine B, (±)-kopsidasine, (±)-kopsidasine-N-oxide, (±)-kopsijasminilam and (±)-11-methoxykopsilongine. Tetrahedron 2002, 17, 3423-3443.
8.De Martino, G.; La Regina, G.; Coluccia, A.; Edler, M. C.; Barbera, M. C.; Brancale, A.; Wilcox, E.; Hamel, E.; Artico, M.; Silvestri, R. Arylthioindoles, potent inhibitors of tubulin polymerization. Journal of Medicinal Chemistry ,2004, 47, 6120-6123.
9.Benington, F.; Morin, R. D.; Clark, L. C.; Jr. Mesclin analogs. VIII. Subtituted 5-methoxy- and 5,6,7-timethoxyindoles. Journal of Medicinal Chemistry 1958, 23, 19-23.
10.N, K.; K, J.; S, D.; PW, B.; RT, C.; S, T.; DH, T.; Jr, K. J.; C, N.-N.; A, S.; R, Y.-S.; Q., W. Synthesis and biological evaluation of quinoline salicylic acids as P-selectin antagonists. Jounal of Medicinal Chemistry 2007 , 50, 21-39
11.Crestini, C.; Saladino, R. A new efficient and mild syntheisis of 2-oxindoles by one-pot Wolff-kishner like reduction of isatin derivatives. synthetic communications 1994, 24, 2835-2841.
12.OLgen, S.; GOTZ, C.; Jose, J. Synthesis and biological evaluation of 3-(substituted-benzylidene)-1,3-dihydro-indolin Derivatives as Human Protein Kinase CK2 and p60c-Sre tyrosine kinase inhibitors. Biological & Parmaceutical Bulletin, 2007, 30, 715-718.

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系統識別號 U0007-1307200917205300
論文名稱(中文) 微生物轉換ent-13-Hydroxy-kaur-15-en-19-oic acid
論文名稱(英文) Microbial Transformation of ent-13-Hydroxy-kaur-15-en-19-oic acid
校院名稱 臺北醫學大學
系所名稱(中) 藥學研究所
系所名稱(英) Graduate Institute of Pharmacy
學年度 97
學期 2
出版年 98
研究生(中文) 蔡亞恩
學號 M301096009
學位類別 碩士
語文別 中文
口試日期 2009-06-23
論文頁數 135頁
口試委員 指導教授-林淑娟
委員-陳國棟
委員-張淑芬
關鍵字(中) 微生物轉換
二萜
關鍵字(英) Microbial transformation
Diterpene
學科別分類
中文摘要 微生物轉換是利用微生物細胞中的酵素系統進行構造修飾化合物,因此也具有酵素反應的立體、部位專一特性,且反應發生於溫和環境。Stevioside (1)為天然的甜味劑,可自Stevia rebaudiana (Bertoni) Bertoni的葉子萃取得到,而stevioside及其相關化合物具有許多藥理活性,例如:降血糖、降血壓、抗發炎、抗癌及免疫調節的功能。以化學觀點來看,具bridged ring架構的diterpenes很適合做為研究微生物轉換的受質,因此許多由stevioside衍生的化合物已用於微生物轉換的研究。ent-13-Hydroxy-kaur-15-en-19-oic acid (3)屬於ent-kaurene diterpene,為stevioside以酸水解所得的其中一個產物,然而目前仍無針對此化合物進行微生物轉換及生物活性試驗之報導。為了產生新的化合物以用於活性試驗,因此利用微生物轉換技術構造修飾3。由菌種篩選發現, Aspergillus niger、Cunninghamella bainieri及Mortierella isabellina可產生各種化合物且具再現性,因此選擇這些菌種進行大量發酵培養,再經由抽取、分離及純化以得到代謝物。研究發現經由Asp. niger代謝得到ent-13,16beta-dihydroxy-kauran-19-oic acid (4)、ent-13-hydroxy-11alpha,16alpha-epoxy-19-oic acid (5)、steviol (6)及ent-7alpha,13-dihydroxy-kaur-15-en-19-oic acid (7);經由C. bainieri 代謝得到7、ent-7alpha,14beta-dihydroxy-16-oxo-beyeran-19-oic acid (8)、ent-14beta-hydroxy-16-oxo-beyeran-19-oic acid (9)、ent-9alpha,13-dihydroxy-kaur-15-en-19-oic acid (10)及ent-13,15beta-dihydroxy-kaur-16-en-19-oic acid (11);經由M. isabellina 得到 5、10、ent-3alpha,13-dihydroxy-kaur-15-en-19-oic acid (12)及ent-13,17-dihydroxy-kaur-15-en-19-oic acid (13),其中7 及9 曾以甲基化衍生物被分離出,而5、8、10及12為新的化合物,所得的新化合物皆經由一維、二維核磁共振光譜,低解析、高解析質譜及X-ray結晶繞射分析確定結構。此外,受質3及分離所得的化合物,其生物活性試驗,目前正在進行中。
英文摘要 Microbial transformation can modify the structure of compounds due to the presence of numerous enzymatic activities in whole cells that act as biocatalysts. The synthesis of optically active compounds by using microbial models has been successfully applied for regio- and stereoselective under mild conditions. Stevioside (1) is a natural sweetener extracted from leaves of Stevia rebaudiana (Bertoni) Bertoni. Stevioside and related compounds have many therapeutic benefits such as anti-hyperglycemic, anti-hypertensive, anti-inflammatory, anti-tumor, and immunomodulatory actions. From a chemical point of view, the bridged ring skeleton of diterpenes is a suitable substrate for the study of microbial transformation. Thus, a number of compounds derived from stevioside (1) have been studied on the microbial transformation. ent-13-Hydroxy-kaur-15-en-19-oic acid (3), an ent-kaurene diterpene, is one of compounds from acid hydrolysis of 1. However, no report on the microbial transformation and biological activity of 3 has been described. In an effort to produce new functionalized compounds for biological testings, the microbial transformation of 3 was carried out. By screening several microorganisms, Aspergillus niger, Cunninghamella bainieri, Mortierella isabellina were selected for preparative-scale transformation of 3 because they reproducibly produced metabolites. Incubation of 3 with Asp. niger produced ent-13,16beta-dihydroxy-kauran-19-oic acid (4), ent-13-hydroxy-11alpha,16alpha-epoxy-kauran-19-oic acid (5), steviol (6), and ent-7alpha,13-dihydroxy-kaur-15-en-19-oic acid (7). Incubation of 3 with C. bainieri produced 7, ent-7alpha,14beta-dihydroxy-16-oxo-beyeran-19-oic acid (8), ent-14beta-hydroxy-16-oxo-beyeran-19-oic acid (9), ent-9alpha,13-dihydroxy-kaur-15-en-19-oic acid (10), and ent-13,15beta-dihydroxy-kaur-16-en-19-oic acid (11). Incubation of 3 with M. isabellina produced 5, 10, ent-3alpha,13-dihydroxy-kaur-15-en-19-oic acid (12), and ent-13,17-dihydroxy-kaur-15-en-19-oic acid (13). Among them, 7 and 9 have been isolated as methylated derivatives. Compounds 5, 8, 10, and 12 are new compounds. The structures of the new compounds were established on the basis of HRESIMS, 1D and 2D NMR, and X-ray crystallographic analyses. The biological testing of these compounds is still in progress.
論文目次 目錄………………………………………………………………………i
附表目錄………………………………………………………………iii
附圖目錄………………………………………………………………iv
流程目錄………………………………………………………………viii
詞彙………………………………………………………………………1
中文摘要…………………………………………………………………2
英文摘要…………………………………………………………………4
第一章 緒論
研究背景…………………………………………………………………6
研究目的…………………………………………………………………18
第二章 結果與討論
受質ent-13-hydroxy-kaur-15-en-19-oic acid (3)之製備與結構判定…………………………………………………………………………24
ent-13-Hydroxy-kaur-15-en-19-oic acid (3)與Aspergillus niger之微生物轉換與代謝物結構判定………………………………………27
ent-13-Hydroxy-kaur-15-en-19-oic acid (3)與Cunninghamella
bainieri之微生物轉換與代謝物結構判定……………………………31
ent-13-Hydroxy-kaur-15-en-19-oic acid (3)與Mortierella
isabellina之微生物轉換與代謝物結構判定…………………………37
結論………………………………………………………………………40
第三章 實驗部分
實驗儀器…………………………………………………………………42
管柱………………………………………………………………………43
溶劑與試劑………………………………………………………………43
微生物……………………………………………………………………45
受質3之製備……………………………………………………………46
微生物轉換實驗
培養及發酵程序…………………………………………………………47
ent-13-Hydroxy-kaur-15-en-19-oic acid (3)之微生物轉換實
驗…………………………………………………………………………50
ent-13-Hydroxy-kaur-15-en-19-oic acid (3)與Aspergillus
niger之微生物轉換……………………………………………………51
ent-13-Hydroxy-kaur-15-en-19-oic acid (3)與
Cunninghamella bainieri之微生物轉換……………………………54
ent-13-Hydroxy-kaur-15-en-19-oic acid (3)與Mortierella
isabellina之微生物轉換………………………………………………58
附錄 產率附表…………………………………………………………64
參考文獻………………………………………………………………129
參考文獻 參考文獻
1. Liese, A.; Seelbach, K.; Wandrey, C. Industrial biotransformations. Wiley-VCH: Germany 2000.
2. 蘇遠志, 應用微生物學. 華香園出版社, 1999.
3. Wackett, L. P.; Hershberger, C. D. Biocatalysis and biodegradation. ASM Press: Washington, D.C. 2001.
4. Asha, S.; Vidyavathi, M. Cunninghamella ?{ A microbial model for drug metabolism studies ?{ A review. Biotechnol. Adv. 2009, 27, 16-29.
5. Faber, K. Biotransformations in organic chemistry, 4th Ed. Springer-Verlag: Germeny, 2000.
6. Keshetty, S.; Ciddi, V. Biotransformation of drugs by microbial cultures for predicting mammalian drug metabolism. Biotechnol. Adv. 2003, 21, 3-39.
7. Venisetty, R. K.; Ciddi, V. Application of microbial biotransformation for the new drug discovery using natural drugs as substrates. Curr. Pharm. Biotechnol. 2003, 4, 153-167.
8. Marsheck, W. J.; Karim, A. Preparation of metabolites of spironolactone by microbial oxygenation. Appl. Microbiol. 1972, 25, 647-649.
9. Smith, R. V.; Rosazza, J. P. Microbial models of mammalian metabolism. J. Nat. Prod. 1983, 46, 79-91.
10. Smith, R. V.; Rosazza, J. P. Microbial models of mammalian metabolism. Aromatic hydroxylatiom. Arch. Biochem. Biophysic. 1974, 161, 551-558.
11. Lacroix, I.; Biton, J.; Azerad, R. Microbial models of drug metabolism: microbial transformations of trimegestone (RU27987), a 3-keto-delta(4,9(10))-19-norsteroid drug. Bioorg. Med. Chem. 1999, 7, 2329-2341.
12. Teo, S. K.; Colburn, W. A.; Tracewell, W. G.; Kook, K. A.; Stirling, D. I.; Jaworsky, M. S.; Scheffler, M. A.; Thomas, S. D.; Laskin, O. L. Clinical pharmacokinetics of thalidomide. Clin. Pharmacokinet. 2004, 43, 311-327.
13. Kardinahl, S.; Rabelt, D.; Reschke, M. Biotransformation: from vision to technology. Chem. Ing. Tech. 2006, 78, 209-217.
14. Tao, J.; Xu, J. H. Biocatalysis in development of green pharmaceutical processes. Curr. Opin. Chem. Biol. 2009, 13, 1-8.
15. Lam, K. S. New aspects of natural products in drug discovery. Trends Microbiol. 2007, 15, 279-289.
16. Lefevre, F.; Robe, P.; Jarrin, C.; Ginolhac, A.; Zago, C.; Auriol, D.; Vogel, T. M.; Simonet, P.; Nalin, R. Drugs from hidden bugs: their discovery via untapped resources. Res. Microbiol. 2008, 159, 153-161.
17. Feher, M.; Schmidt, J. M. Property distributions: differences between drugs, natural products, and molecules from combinatorial chemistry. J. Chem. Inf. Comput. Sci. 2003, 43, 218-227.
18. Ortholand, J. Y.; Ganesan, A. Natural products and combinatorial chemistry: back to the future. Curr. Opin. Chem. Biol. 2004, 8, 271-280.
19. Geuns, J. M. Stevioside. Phytochemistry 2003, 64, 913-921.
20. Kinghorn, A. D.; Soejarto, D. D. Discovery of terpenoid and phenolic sweeteners from plants. Pure Appl. Chem. 2002, 74, 1169-1179.
21. Chatsudthipong, V.; Muanprasat, C. Stevioside and related compounds: Therapeutic benefits beyond sweetness. Pharmacol. Ther. 2009, 121, 41-54.
22. Mosettig, E.; Beglinger, U.; Dolder, F.; Lichiti H.; Quitt P.; Waters J. A. The absolute configuration of steviol and isosteviol. J. Am. Chem. Soc. 1963, 85, 2305-2309
23. Yamasaki, K; Kphada, H; Kobayashi, T.; Kasai, R.; Tanaka, O. Structures of stevia diterpene-glucosides: Application of 13C NMR. Tetrahedron Lett. 1976, 17, 1005-1008.
24. Ferreira, E. B.; de Assis Rocha Neves, F.; da Costa, M. A.; do Prado, W. A.; de Araujo Funari Ferri, L.; Bazotte, R. B. Comparative effects of Stevia rebaudiana leaves and stevioside on glycaemia and hepatic gluconeogenesis. Planta Med. 2006, 72, 691-696.
25. Toskulkao, C.; Sutheerawattananon, M.; Piyachaturawat, P. Inhibitory effect of steviol, a metabolite of stevioside, on glucose absorption in everted hamster intestine in vitro. Toxicol. Lett. 1995, 80, 153-159.
26. Chen, T. H.; Chen, S. C.; Chan, P.; Chu, Y. L.; Yang, H. Y.; Cheng, J. T. Mechanism of the hypoglycemic effect of stevioside, a glycoside of Stevia rebaudiana. Planta Med. 2005, 71, 108-113.
27. Jeppesen, P. B.; Gregersen, S.; Poulsen, C. R.; Hermansen, K. Stevioside acts directly on pancreatic beta cells to secrete insulin: actions independent of cyclic adenosine monophosphate and adenosine triphosphate-sensitive K+-channel activity. Metabolism 2000, 49, 208-214.
28. Melis, M. S.; Sainati, A. R.; Maciel, R. E. Effects of two concentrations of stevioside on renal function and mean arterial pressure in rats. IRCS Med. Sci. 1986, 14, 973.
29. Melis, M. S.; Sainati, A. R. Effect of calcium and verapamil on renal function of rats during treatment with stevioside. J. Ethnopharmacol. 1991, 33, 257-262.
30. Geuns, J. M.; Buyse, J.; Vankeirsbilck, A.; Temme, E. H. Metabolism of stevioside by healthy subjects. Exp. Biol. Med. (Maywood) 2007, 232, 164-173.
31. Boonkaewwan, C.; Toskulkao, C.; Vongsakul, M. Anti-Inflammatory and Immunomodulatory Activities of Stevioside and Its Metabolite Steviol on THP-1 Cells. J. Agric. Food Chem. 2006, 54, 785-789.
32. Sehar, I.; Kaul, A.; Bani, S.; Pal, H. C.; Saxena, A. K. Immune up regulatory response of a non-caloric natural sweetener, stevioside. Chem. Biol. Interact. 2008, 173, 115-121.
33. Yasukawa, K.; Kitanaka, S.; Seo, S. Inhibitory effect of stevioside on tumor promotion by 12-O-tetradecanoylphorbol-13-acetate in two-stage carcinogenesis in mouse skin. Biol. Pharm. Bull. 2002, 25, 1488-1490.
34. Mizushina, Y.; Akihisa, T.; Ukiya, M.; Hamasaki, Y.; Murakami-Nakai, C.; Kuriyama, I.; Takeuchi, T.; Sugawara, F.; Yoshida, H. Structural analysis of isosteviol and related compounds as DNA polymerase and DNA topoisomerase inhibitors. Life Sci. 2005, 77, 2127-2140.
35. Takasaki, M.; Konoshima, T.; Kozuka, M.; Tokuda, H.; Takayasu, J.; Nishino, H.; Miyakoshi, M.; Mizutani, K.; Lee, K.-H. Cancer preventive agents. Part 8: Chemopreventive effects of stevioside and related compounds. Bioorg. Med. Chem. 2009, 17, 600-605.
36. Tomita, T.; Sato, N.; Arai, T.; Shiraishi, H.; Sato, M.; Takeuchi, M.; Kamio, Y. Bactericidal activity of a fermented hot-water extract from Stevia rebaudiana Bertoni towards enterohemorrhagic Escherichia coli O157:H7 and other food-borne pathogenic bacteria. Microbiol. Immunol. 1997, 41, 1005-1009.
37. Takahashi, K.; Matsuda, M.; Ohashi, K.; Taniguchi, K.; Nakagomi, O.; Abe, Y.; Mori, S.; Sato, N.; Okutani, K.; Shigeta, S. Analysis of anti-rotavirus activity of extract from Stevia rebaudiana. Antiviral Res. 2001, 49, 15-24.
38. Shiozaki, K.; Fujii, A.; Nakano, T.; Yamaguchi, T.; Sato, M. Inhibitory effects of hot water extract of the Stevia stem on the contractile response of the smooth muscle of the guinea pig ileum. Biosci. Biotechnol. Biochem. 2006, 70, 489-494.
39. Pezzuto, J. M.; Compadre, C. M.; Swanson, S. M.; Nanayakkara, D.; Kinghorn, A. D. Metabolically activated steviol, the aglycone of stevioside, is mutagenic. Proc. Natl. Acad. Sci. U S A 1985, 82, 2478-2782.
40. Matsui, M.; Sofuni, T.; Nohmi, T. Regionally-targeted mutagenesis by metabolically-activated steviol: DNA sequence analysis of steviol-induced mutants of guanine phosphoribosyltransferase (gpt) gene of Salmonella typhimurium TM677. Mutagenesis 1996, 11, 565-572.
41. Klongpanichpak, S.; Temcharoen, P.; Toskulkao, C.; Apibal, S.; Glinsukon, T. Lack of mutagenicity of stevioside and steviol in Salmonella typhimurium TA 98 and TA 100. J. Med. Assoc. Thai. 1997, 80 Suppl 1, S121-128.
42. Terai, T.; Ren, H.; Mori, G.; Yamaguchi, Y.; Hayashi, T. Mutagenicity of steviol and its oxidative derivatives in Salmonella typhimurium TM677. Chem. Pharm. Bull. 2002, 50, 1007-1010.
43. Planas, G. M.; Kucacute, J. Contraceptive Properties of Stevia rebaudiana. Science 1968, 162, 1007.
44. Shiotsu, S. Fertility study of Stevia decoction in rats. Tech. J. Food Chem. Chemicals. 1996, 4, 108-113.
45. Melis, M. S. Effects of chronic administration of Stevia rebaudiana on fertility in rats. J. Ethnopharmacol. 1999, 67, 157-161.
46. Geuns, J. M.; Bruggeman, V.; Buyse, J. G. Effect of stevioside and steviol on the developing broiler embryos. J. Agric. Food Chem. 2003, 51, 5162-5167.
47. Koyama, E.; Sakai, N.; Ohori, Y.; Kitazawa, K.; Izawa, O.; Kakegawa, K.; Fujino, A.; Ui, M. Absorption and metabolism of glycosidic sweeteners of stevia mixture and their aglycone, steviol, in rats and humans. Food Chem. Toxicol. 2003, 41, 875-883.
48. Roberts, A.; Renwick, A. G. Comparative toxicokinetics and metabolism of rebaudioside A, stevioside, and steviol in rats. Food Chem. Toxicol. 2008, 46 Suppl 7, S31-39.
49. 張正豪, 微生物轉換Steviol及ent-16??-Hydroxybeyeran-19-oic acid. 台北醫學大學藥學院藥學研究所碩士論文, 2002.
50. Avent, A. G.; Hanson, J. R.; De Oliveira, B. H. Hydrolysis of the diterpenoid glycoside, stevioside. Phytochemistry 1990, 29, 2712-2715.
51. Hanson, J. R.; De Oliveira, B. H. The microbiological transformation of steviol derivatives by Rhizopus stolinifer and Gibberella fujikuroi. Phytochemistry 1990, 29, 3805-3807.
52. Hsu, F. L.; Hou, C. C.; Yang, L. M.; Cheng, J. T.; Chi, T. C.; Liu, P. C.; Lin, S. J. Microbial transformations of isosteviol. J. Nat. Prod. 2002, 65, 273-277.
53. Chang, S. F.; Yang, L. M.; Hsu, F. L.; Hsu, J. Y.; Liaw, J. H.; Lin, S. J. Transformation of steviol-16??,17-epoxide by Streptomyces griseus and Cunninghamella bainieri. J. Nat. Prod. 2006, 69, 1450- 1455.
54. Yang, L. M.; Hsu, F. L.; Chang, S. F.; Cheng, J. T.; Hsu, J. Y.; Hsu, C. Y.; Liu, P. C.; Lin, S. J. Microbial metabolism of steviol and steviol-16??,17-epoxide. Phytochemistry 2007, 68, 562-570.
55. Chang, S. F.; Yang, L. M.; Lo, C. H.; Liaw, J. H.; Wang, L. H.; Lin, S. J. Microbial transformation of isosteviol and bioactivities against the glucocorticoid/androgen response elements. J. Nat. Prod. 2008, 71, 87-92.
56. Chou, B. H.; Yang, L. M.; Chang, S. F.; Hsu, F. L.; Lo, C. H.; Liaw, J. H.; Liu, P. C.; Lin, S. J. Microbial transformation of isosteviol lactone and evaluation of the transformation products on androgen response element. J. Nat. Prod. 2008, 71, 602-607.
57. Avent, A. G.; Hanson, J. R.; Hitchcock, P. B.; De Oliveira, B. H. The influence of a 15-hydroxy group on the rearrangement reactions of steviol and its 16,17-epoxide. J. Chem. Soc., Perkin Trans. 1 1990, 2661-2665.
58. 許如瑩, 微生物轉換Steviol-16??,17-epoxide. 台北醫學大學藥學系碩士論文, 2005.
59. Ohtani, K.; Aikawa, Y.; Kasai, R.; Chou, W.-H.; Yamasaki, z.; Tanaka, O. Minor diterpene glycosides from sweet leaves of Rubus suavissinus. Phytochemistry 1992, 31, 1553-1559.
60. Hanson, J. R.; Hitchcock, P. B.; Takahashi, J. A. Biotransformation of ent-16??,19-dihydroxykaurane by Cephalosporium aphidicola. Phytochemistry 1995, 40, 797-800.
61. Fraga, B. M.; Gonzalez, P.; Hernandez, M. G.; Suarez, S. Biotransformation of 7-oxo-ent-kaur-16-ene derivatives by Gibberella fujikuroi. Tetrahedron 2005, 61, 5623-5632.
62. Fraga, B. M.; Gonzalez, P.; Guillermo, R.; Hernandez, M. G. A study of the microbiological reduction of ??,??-unsaturated carbonyl ent-kaurenes by Gibberella fujikuroi. Tetrahedron 1996, 52, 13767-13782.
63. Fraga, B. M.; Hernandez, M. G.; Garcia-Tellado, F.; Gonzalez, P.; Perales, A. The biotransformation of two ent-15??,16??- epoxy-
kaurane derivatives by Gibberella fujikuroi. Phytochemistry 1993, 34, 133-138.
64. Fraga, B. M.; Garcia-Tellado, F.; Gonzalez, P.; Hernandez, M. G. The microbiological transformation of 14??,19-dihydroxy-ent-kaur-15-ene by Gibberella fujikuroi. Phytochemistry 1993, 34, 1035-1040.
65. Fraga, B. M.; Hernandez, M. G.; Gonzalez, P. Biotransformation of two ent-15??-hydroxy-kaur-16-ene derivatives by Gibberella fujikuroi. Phytochemistry 1992, 31, 3845-3849.
66. Birladeanu, L. The story of the Wagner-Meerwein rearrangement. J. Chem. Educ. 2000, 77, 858-863.
67. Chou, B. H.; Yang, L. M.; Chang, S. F.; Hsu, F. L.; Lo, C. H.; Lin, W. K.; Wang, L. H.; Liu, P. C.; Lin, S. J. Fungal transformation of isosteviol lactone and its biological evaluation for inhibiting the AP-1 transcription factor. Phytochemistry 2009, 759-764.

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系統識別號 U0007-2007200900452400
論文名稱(中文) p53和MDM2基因多型性與EGFR和K-ras基因突變在肺癌之間的關聯性
論文名稱(英文) The Relationship Between p53 and MDM2 Polymorphisms With EGFR and K-ras Mutations in Lung Cancer
校院名稱 臺北醫學大學
系所名稱(中) 藥學研究所
系所名稱(英) Graduate Institute of Pharmacy
學年度 97
學期 2
出版年 98
研究生(中文) 林育弘
學號 M301096035
學位類別 碩士
語文別 中文
口試日期 2009-07-07
論文頁數 74頁
口試委員 指導教授-劉興璟
共同指導教授-陳香吟
委員-黃耀斌
委員-方嘉佑
委員-李仁愛
關鍵字(中) 基因多型性;肺癌;p53;MDM2;EGFR;K-ras
關鍵字(英) Polymorphisms;Lung Cancer;p53;MDM2;EGFR;K-ras
學科別分類
中文摘要 肺癌是世界各地造成癌症死因的最主要原因,在台灣亦同。目前已知肺癌的危險因子有很多,包括吸菸、環境煙草煙霧、荷爾蒙、病毒和基因變異。許多研究指出p53和Murine double minute 2(MDM2)的某些基因型和增加癌症風險有關係。同時,Epidermal Growth Factor Receptor(EGFR)和v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog(K-ras)基因突變的分布在肺癌特定族群中有著極大的差異性存在。台灣目前尚未有相關研究在p53和MDM2基因多型性與EGFR和K-ras基因突變和肺癌之間的關聯性。本研究目的是比較肺癌和對照組間之p53 codon 72和MDM2 SNP309基因型的分布,並探討p53和MDM2基因多型性與EGFR和K-ras基因突變在肺癌之間的關聯性。本研究共納入170名肺癌病患和340名正常對照組。以聚合酶連鎖反應-限制酶切割片段長度多型性(PCR-RFLP)的方式分析病患及對照組的週邊血液檢體,檢測p53 codon 72(包括Arg/Arg、Arg/Pro和Pro/Pro三種基因型)和MDM2 SNP309(包括T/T、T/G和G/G三種基因型)的基因變異,並回溯病患的病歷相關資料。研究結果顯示,p53 codon 72和MDM2 SNP309基因型分布在對照組和肺癌組之間無統計上顯著的差異(p值分別為0.35和0.72)。然而,我們發現在不吸菸之女性肺腺癌族群中,EGFR基因突變者較容易有MDM2 SNP309 G對偶基因(Odds Ratio = 2.71;95% CI = 1.10 – 6.69;p值為 0.028)。為了進一步了解肺癌中的基因因子,應該需要更進一步的研究來證實p53和MDM2基因多型性與EGFR和K-ras基因突變在肺癌之間的關聯性和機轉。
英文摘要 Lung cancer is the leading cause of cancer death worldwide. Similarly, it is the leading cause of cancer death in Taiwan. Risk factors for lung cancer include smoking, environmental tobacco smoke(ETS), hormonal, rival and genetic factors. Numbers of studies reported that some genotypes of p53 and Murine double minute 2(MDM2)were potentially associated with an increased risk of cancer. Furthermore, there exists a striking difference in the distribution of Epidermal growth factor receptor(EGFR)and v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog(K-ras) mutation in certain subgroups of lung cancer patients. However, there have been no study performed to explore the relationship between p53 and MDM2 polymorphisms, EGFR and K-ras mutations in lung cancer patients in Taiwan.
The purposes of this study were to compare the distribution of p53 codon 72 and MDM2 SNP309 genotypes between lung cancer patients and normal subjects in the Taiwanese population, and to investigate the relationship between p53 and MDM2 polymorphisms with EGFR and K-ras mutations in lung cancer.
This study included 170 lung cancer patients and 340 normal controls. Genomic DNA from peripheral blood samples was analyzed by polymerase chain reactions – restriction fragment length polymorphism(PCR-RFLP)for genetic variation in p53 codon 72 (Arg/Arg, Arg/Pro, Pro/Pro) and MDM2 SNP309 (T/T, T/G, G/G). The results showed that distribution of p53 codon 72 and MDM2 SNP309 genotypes were not significantly different between overall lung cancer and controls(p = 0.35 and p = 0.72, respectively). However, it was observed that the presence of EGFR mutation was associated with MDM2 SNP309 G allele in the non-smoker female with adenocarincoma subgroup (Odds Ratio = 2.71;95% CI = 1.10 – 6.69;p = 0.028). Further studies are warranted to investigate the relationship and mechanism between p53 and MDM2 polymorphisms with EGFR and K-ras mutations in lung cancer.
論文目次 誌謝 I
中文摘要 II
英文摘要 III
目次 IV
圖目錄 VI
表目錄 VII
第1章 緒論 1
第2章 文獻探討 2
2.1 肺癌概述 2
2.1.1 肺癌簡介 2
2.1.2 肺癌的治療 5
2.1.3 肺癌的危險因子 7
2.2 抑癌基因p53 12
2.2.1 p53基因多型性 13
2.2.2 p53 codon 72基因多型性 14
2.2.3 p53 codon 72基因多型性與癌症之關係 14
2.3 致癌基因MDM2 16
2.3.1 p53和MDM2之關係 16
2.3.2 MDM2 SNP309基因多型性與癌症之關係 17
2.4 肺癌中分子特徵的變異 19
2.4.1 肺癌中p53基因突變 19
2.4.2 肺癌中EGFR與K-ras基因突變 20
第3章 研究目的 24
第4章 研究方法 25
4.1 研究設計 25
4.1.1 肺癌組與對照組之描述 25
4.1.2 病人資料收集 26
4.2 基因型檢測 27
4.2.1 p53和MDM2基因相關之定義 28
4.2.2 DNA的萃取 29
4.2.3 聚合酶連鎖反應-限制酶切割片段長度多型性 30
4.2.4 基因型的判定 33
4.3 統計方法 36
第5章 研究結果 37
5.1 基本資料 37
5.2 基因型分布 39
5.3 次族群分析(Subgroup Analysis) 42
5.3.1 肺腺癌基因型分析 42
5.3.2 年齡分析(以65歲為分層) 45
5.3.3 肺癌嚴重度分析 46
5.4 MDM2 SNP309和p53 codon 72基因型之合併分析 46
5.5 肺癌與吸菸之間的關係 46
5.6 p53及MDM2基因多型性與EGFR基因突變之間的關係 46
5.7 p53及MDM2基因多型性與K-ras基因突變之間的關係 46
第6章 討論 46
6.1 MDM2基因多型性與肺癌之關係 46
6.2 p53基因多型性與肺癌之關係 46
6.3 MDM2基因多型性與EGFR基因突變之關係 46
6.4 MDM2和p53基因型之種族差異 46
6.4.1 對照組分析 46
6.4.2 肺癌病人組分析 46
6.5 研究限制及未來改進方向 46
第7章 結論 46
參考文獻 46

參考文獻 1. Society AC. Global Cancer Facts & Figures 2007. 2007.

2. Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, Thun MJ. Cancer statistics, 2008. CA Cancer J Clin 2008; 58(2): 71-96.

3. 行政院衛生署. 國人96年死因統計 http://www.doh.gov.tw/CHT2006/index_populace.aspx. 2007.

4. Herbst RS, Heymach JV, Lippman SM. Lung cancer. N Engl J Med 2008; 359(13): 1367-1380.

5. Ginsberg MS, Grewal RK, Heelan RT. Lung cancer. Radiol Clin North Am 2007; 45(1): 21-43.

6. Brambilla E, Travis WD, Colby TV, Corrin B, Shimosato Y. The new World Health Organization classification of lung tumours. Eur Respir J 2001; 18(6): 1059-1068.

7. 行政院衛生署國民健康局. 癌症登記報告 Cancer Registry Annual Report, 2006 Taiwan. 2009.

8. Uptodate. Uptodate http://www.uptodateonline.com/home/index.html. 2009.

9. Karnofsky D, Abelmann, W, Craver, L, Burchenal, J. The use of nitrogen mustard in the palliative treatment of cancer. Cancer 1948; 1:634 1948.

10. Buccheri G, Ferrigno D, Tamburini M. Karnofsky and ECOG performance status scoring in lung cancer: a prospective, longitudinal study of 536 patients from a single institution. Eur J Cancer 1996; 32A(7): 1135-1141.

11. Hecht SS. Cigarette smoking and lung cancer: chemical mechanisms and approaches to prevention. Lancet Oncol 2002; 3(8): 461-469.

12. Hecht SS. Tobacco carcinogens, their biomarkers and tobacco-induced cancer. Nat Rev Cancer 2003; 3(10): 733-744.

13. Godtfredsen NS, Prescott E, Osler M. Effect of smoking reduction on lung cancer risk. JAMA 2005; 294(12): 1505-1510.

14. Sun S, Schiller JH, Gazdar AF. Lung cancer in never smokers--a different disease. Nat Rev Cancer 2007; 7(10): 778-790.

15. Lam WK. Lung cancer in Asian women-the environment and genes. Respirology 2005; 10(4): 408-417.

16. Lam WK, White NW, Chan-Yeung MM. Lung cancer epidemiology and risk factors in Asia and Africa. Int J Tuberc Lung Dis 2004; 8(9): 1045-1057.

17. Chiang TA, Wu PF, Wang LF, Lee H, Lee CH, Ko YC. Mutagenicity and polycyclic aromatic hydrocarbon content of fumes from heated cooking oils produced in Taiwan. Mutat Res 1997; 381(2): 157-161.

18. Chiang TA, Wu PF, Ko YC. Identification of carcinogens in cooking oil fumes. Environ Res 1999; 81(1): 18-22.

19. Stayner L, Bena J, Sasco AJ, Smith R, Steenland K, Kreuzer M, Straif K. Lung cancer risk and workplace exposure to environmental tobacco smoke. Am J Public Health 2007; 97(3): 545-551.

20. Beutner KR, Tyring S. Human papillomavirus and human disease. Am J Med 1997; 102(5A): 9-15.

21. Tsuhako K, Nakazato I, Hirayasu T, Sunakawa H, Iwamasa T. Human papillomavirus DNA in adenosquamous carcinoma of the lung. J Clin Pathol 1998; 51(10): 741-749.

22. Cheng YW, Wu MF, Wang J, Yeh KT, Goan YG, Chiou HL, Chen CY, Lee H. Human papillomavirus 16/18 E6 oncoprotein is expressed in lung cancer and related with p53 inactivation. Cancer Res 2007; 67(22): 10686-10693.

23. Cheng YW, Chiou HL, Sheu GT, Hsieh LL, Chen JT, Chen CY, Su JM, Lee H. The association of human papillomavirus 16/18 infection with lung cancer among nonsmoking Taiwanese women. Cancer Res 2001; 61(7): 2799-2803.

24. Fei Y, Yang J, Hsieh WC, Wu JY, Wu TC, Goan YG, Lee H, Cheng YW. Different human papillomavirus 16/18 infection in Chinese non-small cell lung cancer patients living in Wuhan, China. Jpn J Clin Oncol 2006; 36(5): 274-279.

25. Wu CT, Chang YL, Shih JY, Lee YC. The significance of estrogen receptor beta in 301 surgically treated non-small cell lung cancers. J Thorac Cardiovasc Surg 2005; 130(4): 979-986.

26. Yager JD, Davidson NE. Estrogen carcinogenesis in breast cancer. N Engl J Med 2006; 354(3): 270-282.

27. Matakidou A, Eisen T, Houlston RS. Systematic review of the relationship between family history and lung cancer risk. Br J Cancer 2005; 93(7): 825-833.

28. Cote ML, Kardia SL, Wenzlaff AS, Ruckdeschel JC, Schwartz AG. Risk of lung cancer among white and black relatives of individuals with early-onset lung cancer. JAMA 2005; 293(24): 3036-3042.

29. Haiman CA, Stram DO, Wilkens LR, Pike MC, Kolonel LN, Henderson BE, Le Marchand L. Ethnic and racial differences in the smoking-related risk of lung cancer. N Engl J Med 2006; 354(4): 333-342.

30. Schwartz AG, Prysak GM, Bock CH, Cote ML. The molecular epidemiology of lung cancer. Carcinogenesis 2007; 28(3): 507-518.

31. Hung RJ, Boffetta P, Brockmoller J, Butkiewicz D, Cascorbi I, Clapper ML, Garte S, Haugen A, Hirvonen A, Anttila S, Kalina I, Le Marchand L, London SJ, Rannug A, Romkes M, Salagovic J, Schoket B, Gaspari L, Taioli E. CYP1A1 and GSTM1 genetic polymorphisms and lung cancer risk in Caucasian non-smokers: a pooled analysis. Carcinogenesis 2003; 24(5): 875-882.

32. Vogelstein B, Lane D, Levine AJ. Surfing the p53 network. Nature 2000; 408(6810): 307-310.

33. Vousden KH, Lane DP. p53 in health and disease. Nat Rev Mol Cell Biol 2007; 8(4): 275-283.

34. Jin S, Levine AJ. The p53 functional circuit. J Cell Sci 2001; 114(Pt 23): 4139-4140.

35. Shu KX, Li B, Wu LX. The p53 network: p53 and its downstream genes. Colloids Surf B Biointerfaces 2007; 55(1): 10-18.

36. Robles AI, Linke SP, Harris CC. The p53 network in lung carcinogenesis. Oncogene 2002; 21(45): 6898-6907.

37. Harris SL, Levine AJ. The p53 pathway: positive and negative feedback loops. Oncogene 2005; 24(17): 2899-2908.

38. Thomas M, Kalita A, Labrecque S, Pim D, Banks L, Matlashewski G. Two polymorphic variants of wild-type p53 differ biochemically and biologically. Mol Cell Biol 1999; 19(2): 1092-1100.

39. Matlashewski GJ, Tuck S, Pim D, Lamb P, Schneider J, Crawford LV. Primary structure polymorphism at amino acid residue 72 of human p53. Mol Cell Biol 1987; 7(2): 961-963.

40. Dumont P, Leu JI, Della Pietra AC, 3rd, George DL, Murphy M. The codon 72 polymorphic variants of p53 have markedly different apoptotic potential. Nat Genet 2003; 33(3): 357-365.

41. Storey A, Thomas M, Kalita A, Harwood C, Gardiol D, Mantovani F, Breuer J, Leigh IM, Matlashewski G, Banks L. Role of a p53 polymorphism in the development of human papillomavirus-associated cancer. Nature 1998; 393(6682): 229-234.

42. Howie HL, Katzenellenbogen RA, Galloway DA. Papillomavirus E6 proteins. Virology 2009; 384(2): 324-334.

43. Szymanowska A, Jassem E, Dziadziuszko R, Borg A, Limon J, Kobierska-Gulida G, Rzyman W, Jassem J. Increased risk of non-small cell lung cancer and frequency of somatic TP53 gene mutations in Pro72 carriers of TP53 Arg72Pro polymorphism. Lung Cancer 2006; 52(1): 9-14.

44. Shen H, Zheng Y, Sturgis EM, Spitz MR, Wei Q. P53 codon 72 polymorphism and risk of squamous cell carcinoma of the head and neck: a case-control study. Cancer Lett 2002; 183(2): 123-130.

45. Zhang X, Miao X, Guo Y, Tan W, Zhou Y, Sun T, Wang Y, Lin D. Genetic polymorphisms in cell cycle regulatory genes MDM2 and TP53 are associated with susceptibility to lung cancer. Hum Mutat 2006; 27(1): 110-117.

46. Phang BH, Linn YC, Li H, Sabapathy K. MDM2 SNP309 G allele decreases risk but does not affect onset age or survival of Chinese leukaemia patients. Eur J Cancer 2008; 44(5): 760-766.

47. Riley T, Sontag E, Chen P, Levine A. Transcriptional control of human p53-regulated genes. Nat Rev Mol Cell Biol 2008; 9(5): 402-412.

48. Iwakuma T, Lozano G. MDM2, an introduction. Mol Cancer Res 2003; 1(14): 993-1000.

49. Moll UM, Petrenko O. The MDM2-p53 interaction. Mol Cancer Res 2003; 1(14): 1001-1008.

50. Chene P. Inhibiting the p53-MDM2 interaction: an important target for cancer therapy. Nat Rev Cancer 2003; 3(2): 102-109.

51. Deb SP. Cell cycle regulatory functions of the human oncoprotein MDM2. Mol Cancer Res 2003; 1(14): 1009-1016.

52. Freedman DA, Wu L, Levine AJ. Functions of the MDM2 oncoprotein. Cell Mol Life Sci 1999; 55(1): 96-107.

53. Whibley C, Pharoah PD, Hollstein M. p53 polymorphisms: cancer implications. Nat Rev Cancer 2009; 9(2): 95-107.

54. Momand J, Jung D, Wilczynski S, Niland J. The MDM2 gene amplification database. Nucleic Acids Res 1998; 26(15): 3453-3459.

55. Bond GL, Hu W, Bond EE, Robins H, Lutzker SG, Arva NC, Bargonetti J, Bartel F, Taubert H, Wuerl P, Onel K, Yip L, Hwang SJ, Strong LC, Lozano G, Levine AJ. A single nucleotide polymorphism in the MDM2 promoter attenuates the p53 tumor suppressor pathway and accelerates tumor formation in humans. Cell 2004; 119(5): 591-602.

56. Hong Y, Miao X, Zhang X, Ding F, Luo A, Guo Y, Tan W, Liu Z, Lin D. The role of P53 and MDM2 polymorphisms in the risk of esophageal squamous cell carcinoma. Cancer Res 2005; 65(20): 9582-9587.

57. Cattelani S, Defferrari R, Marsilio S, Bussolari R, Candini O, Corradini F, Ferrari-Amorotti G, Guerzoni C, Pecorari L, Menin C, Bertorelle R, Altavista P, McDowell HP, Boldrini R, Dominici C, Tonini GP, Raschella G, Calabretta B. Impact of a single nucleotide polymorphism in the MDM2 gene on neuroblastoma development and aggressiveness: results of a pilot study on 239 patients. Clin Cancer Res 2008; 14(11): 3248-3253.

58. Ohmiya N, Taguchi A, Mabuchi N, Itoh A, Hirooka Y, Niwa Y, Goto H. MDM2 promoter polymorphism is associated with both an increased susceptibility to gastric carcinoma and poor prognosis. J Clin Oncol 2006; 24(27): 4434-4440.

59. Hirata H, Hinoda Y, Kikuno N, Kawamoto K, Suehiro Y, Tanaka Y, Dahiya R. MDM2 SNP309 polymorphism as risk factor for susceptibility and poor prognosis in renal cell carcinoma. Clin Cancer Res 2007; 13(14): 4123-4129.

60. Park SH, Choi JE, Kim EJ, Jang JS, Han HS, Lee WK, Kang YM, Park JY. MDM2 309T>G polymorphism and risk of lung cancer in a Korean population. Lung Cancer 2006; 54(1): 19-24.

61. Hu W, Feng Z, Ma L, Wagner J, Rice JJ, Stolovitzky G, Levine AJ. A single nucleotide polymorphism in the MDM2 gene disrupts the oscillation of p53 and MDM2 levels in cells. Cancer Res 2007; 67(6): 2757-2765.

62. Bond GL, Hirshfield KM, Kirchhoff T, Alexe G, Bond EE, Robins H, Bartel F, Taubert H, Wuerl P, Hait W, Toppmeyer D, Offit K, Levine AJ. MDM2 SNP309 accelerates tumor formation in a gender-specific and hormone-dependent manner. Cancer Res 2006; 66(10): 5104-5110.

63. Vahakangas KH, Bennett WP, Castren K, Welsh JA, Khan MA, Blomeke B, Alavanja MC, Harris CC. p53 and K-ras mutations in lung cancers from former and never-smoking women. Cancer Res 2001; 61(11): 4350-4356.

64. Toyooka S, Tsuda T, Gazdar AF. The TP53 gene, tobacco exposure, and lung cancer. Hum Mutat 2003; 21(3): 229-239.

65. Gealy R, Zhang L, Siegfried JM, Luketich JD, Keohavong P. Comparison of mutations in the p53 and K-ras genes in lung carcinomas from smoking and nonsmoking women. Cancer Epidemiol Biomarkers Prev 1999; 8(4 Pt 1): 297-302.

66. Shigematsu H, Lin L, Takahashi T, Nomura M, Suzuki M, Wistuba, II, Fong KM, Lee H, Toyooka S, Shimizu N, Fujisawa T, Feng Z, Roth JA, Herz J, Minna JD, Gazdar AF. Clinical and biological features associated with epidermal growth factor receptor gene mutations in lung cancers. J Natl Cancer Inst 2005; 97(5): 339-346.

67. Jang TW, Oak CH, Chang HK, Suo SJ, Jung MH. EGFR and KRAS mutations in patients with adenocarcinoma of the lung. Korean J Intern Med 2009; 24(1): 48-54.

68. Kosaka T, Yatabe Y, Endoh H, Kuwano H, Takahashi T, Mitsudomi T. Mutations of the epidermal growth factor receptor gene in lung cancer: biological and clinical implications. Cancer Res 2004; 64(24): 8919-8923.

69. Tam IY, Chung LP, Suen WS, Wang E, Wong MC, Ho KK, Lam WK, Chiu SW, Girard L, Minna JD, Gazdar AF, Wong MP. Distinct epidermal growth factor receptor and KRAS mutation patterns in non-small cell lung cancer patients with different tobacco exposure and clinicopathologic features. Clin Cancer Res 2006; 12(5): 1647-1653.

70. Ciardiello F, Tortora G. EGFR antagonists in cancer treatment. N Engl J Med 2008; 358(11): 1160-1174.

71. Yarden Y, Sliwkowski MX. Untangling the ErbB signalling network. Nat Rev Mol Cell Biol 2001; 2(2): 127-137.

72. Sharma SV, Bell DW, Settleman J, Haber DA. Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer 2007; 7(3): 169-181.

73. Pao W, Miller VA. Epidermal growth factor receptor mutations, small-molecule kinase inhibitors, and non-small-cell lung cancer: current knowledge and future directions. J Clin Oncol 2005; 23(11): 2556-2568.

74. Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW, Harris PL, Haserlat SM, Supko JG, Haluska FG, Louis DN, Christiani DC, Settleman J, Haber DA. Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med 2004; 350(21): 2129-2139.

75. Paez JG, Janne PA, Lee JC, Tracy S, Greulich H, Gabriel S, Herman P, Kaye FJ, Lindeman N, Boggon TJ, Naoki K, Sasaki H, Fujii Y, Eck MJ, Sellers WR, Johnson BE, Meyerson M. EGFR mutations in lung cancer: correlation with clinical response to gefitinib therapy. Science 2004; 304(5676): 1497-1500.

76. Pao W, Miller V, Zakowski M, Doherty J, Politi K, Sarkaria I, Singh B, Heelan R, Rusch V, Fulton L, Mardis E, Kupfer D, Wilson R, Kris M, Varmus H. EGF receptor gene mutations are common in lung cancers from "never smokers" and are associated with sensitivity of tumors to gefitinib and erlotinib. Proc Natl Acad Sci U S A 2004; 101(36): 13306-13311.

77. Shigematsu H, Gazdar AF. Somatic mutations of epidermal growth factor receptor signaling pathway in lung cancers. Int J Cancer 2006; 118(2): 257-262.

78. Downward J. Targeting RAS signalling pathways in cancer therapy. Nat Rev Cancer 2003; 3(1): 11-22.

79. Pao W, Wang TY, Riely GJ, Miller VA, Pan Q, Ladanyi M, Zakowski MF, Heelan RT, Kris MG, Varmus HE. KRAS mutations and primary resistance of lung adenocarcinomas to gefitinib or erlotinib. PLoS Med 2005; 2(1): e17.

80. Kosaka T, Yatabe Y, Onozato R, Kuwano H, Mitsudomi T. Prognostic implication of EGFR, KRAS, and TP53 gene mutations in a large cohort of Japanese patients with surgically treated lung adenocarcinoma. J Thorac Oncol 2009; 4(1): 22-29.

81. Hu Z, Ma H, Lu D, Qian J, Zhou J, Chen Y, Xu L, Wang X, Wei Q, Shen H. Genetic variants in the MDM2 promoter and lung cancer risk in a Chinese population. Int J Cancer 2006; 118(5): 1275-1278.

82. Wang YC, Chen CY, Chen SK, Chang YY, Lin P. p53 codon 72 polymorphism in Taiwanese lung cancer patients: association with lung cancer susceptibility and prognosis. Clin Cancer Res 1999; 5(1): 129-134.

83. Bond GL, Levine AJ. A single nucleotide polymorphism in the p53 pathway interacts with gender, environmental stresses and tumor genetics to influence cancer in humans. Oncogene 2007; 26(9): 1317-1323.

84. Liu G, Wheatley-Price P, Zhou W, Park S, Heist RS, Asomaning K, Wain JC, Lynch TJ, Su L, Christiani DC. Genetic polymorphisms of MDM2, cumulative cigarette smoking and nonsmall cell lung cancer risk. Int J Cancer 2008; 122(4): 915-918.

85. Kuroda Y, Nakao H, Ikemura K, Katoh T. Association between the TP53 codon72 polymorphism and oral cancer risk and prognosis. Oral Oncol 2007; 43(10): 1043-1048.

86. Sotamaa K, Liyanarachchi S, Mecklin JP, Jarvinen H, Aaltonen LA, Peltomaki P, de la Chapelle A. p53 codon 72 and MDM2 SNP309 polymorphisms and age of colorectal cancer onset in Lynch syndrome. Clin Cancer Res 2005; 11(19 Pt 1): 6840-6844.

87. Miller DP, Liu G, De Vivo I, Lynch TJ, Wain JC, Su L, Christiani DC. Combinations of the variant genotypes of GSTP1, GSTM1, and p53 are associated with an increased lung cancer risk. Cancer Res 2002; 62(10): 2819-2823.

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系統識別號 U0007-2007200912492900
論文名稱(中文) 尿苷雙磷酸葡萄糖醛酸基轉移酶1A7(UGT1A7)基因型與肺癌之關聯研究
論文名稱(英文) The Correlation between UDP-Glucuronosyltransferase 1A7 (UGT1A7) Genotypes and Lung Cancer
校院名稱 臺北醫學大學
系所名稱(中) 藥學研究所
系所名稱(英) Graduate Institute of Pharmacy
學年度 97
學期 2
出版年 98
研究生(中文) 黃薇伊
學號 M301096021
學位類別 碩士
語文別 中文
口試日期 2009-06-18
論文頁數 82頁
口試委員 委員-方嘉佑
委員-李仁愛
指導教授-陳香吟
關鍵字(中)

雙磷酸葡萄糖
醛酸基轉移酶
1A7
基因型
肺癌
關鍵字(英) UGT1A7
Genotypes
Lung cancer
學科別分類
中文摘要 肺癌是台灣地區癌症死亡原因之首。活化的致癌物質在體內使得某些重要致癌基因發生突變而失去正常調控, 最終形成肺癌。UGT1A7 (UDP-glucuronosyltransferases1A7) 分布於人體肝臟外的組織,負責許多肺癌致癌物質的代謝解毒。UGT1A7 具有基因多型性,許多研究顯示帶有基因變異的UGT1A7酵素代謝能力較野生型酵素差,因此被認為是造成癌症的可能危險因子。本研究的目的在於探討UGT1A7 基因多型性與台灣族群中肺癌的關係。此外,由於肺癌相關致癌基因KRAS 之突變與抽菸高度相關,我們也進一步假設UGT1A7 基因多
型會與發生此突變的風險有關。本研究收納210 位肺癌病人及210 位依性別年齡配對之健康對照組,以PCR-RFLP 方法分析其UGT1A7 基因多型性,另外可取得腫瘤組織DNA 的病人亦同時檢測KRAS 基因codon 12 與13 位置的突變。結果發現Intermediate-activity (UGT1A7*1/*2, UGT1A7*1/*3, UGT1A7*1/*4, UGT1A7*2/*2,
UGT1A7*2/*3)和Low-activity (UGT1A7*3/*3, UGT1A7*4/*4) 兩種代謝能力較低之UGT1A7 基因表現型為肺癌的可能危險因子,危險對比值分別為1.799, 95% CI:1.178-2.748, p=0.006 及 3.333, 95% CI: 1.192-9.322, p=0.032。此增加之罹癌風險在男性有顯著統計學上差異,但在女性則無此發現。另外,上述代謝能力較低之UGT1A7 基因表現型與EGFR 原型之肺癌相關,而與帶有EGFR 突變之肺癌無關。本研究中並未發現UGT1A7 基因多型性與KRAS 基因突變的發生具有顯著之相關性。本研究結果顯示代謝能力較低之UGT1A7 基因表現型為肺癌的可能危險因子。未來應進一步研究受此酵素能力缺損影響的易感族群及其腫瘤上的分子特徵,並且探索其他可能造成致癌基因突變的危險因子,以期對肺癌致病機轉能有更深的認識。
英文摘要 Lung cancer is the leading cause of cancer deaths in Taiwan. The carcinogenesis process involves activated carcinogens which lead to mutations of crucial oncogenes
resulting in tumor development. UGT1A7 (UDP-glucuronosyltransferases1A7) is an important extrahepatic enzyme that detoxifies a variety of lung carcinogens. Genetic polymorphisms in UGT1A7 were shown to have decreased catalytic activity when compared to the wild-type protein and therefore implicated as a cancer risk factor. The purpose of this study was to investigate the association between genetic polymorphisms of UGT1A7 gene and lung cancer in the Taiwanese population. In addition, since KRAS mutation is highly associated with tobacco smoking, we further hypothesized that UGT1A7 polymorphisms might be correlated with the risk of developing this mutation.
The 210 lung cancer patients and 210 healthy individuals enrolled in this matched case-control study were genotyped for UGT1A7 polymorphisms using PCR-RFLP
method. Tumor tissues available from 150 patients were also tested for KRAS codon 12 and 13 mutations. Predicted intermediate-activity UGT1A7 genotypes (UGT1A7*1/*2,
UGT1A7*1/*3, UGT1A7*1/*4, UGT1A7*2/*2, UGT1A7*2/*3) and low-activity UGT1A7 genotypes (UGT1A7*3/*3, UGT1A7*4/*4) were both significantly correlated with lung cancer risk (p=0.006, odds ratio (OR): 1.799, 95% confidence interval (CI): 1.178-2.748 and p=0.032, OR: 3.333, 95% CI: 1.192-9.322, respectively). Interestingly, the risks were significant in males but not in females. Besides, those lower-activity UGT1A7 genotypes were significantly associated with lung cancer with wild-type EGFR but not with those with EGFR mutations. Of the 150 patients screened for KRAS mutations, 26 (17.3%) patients were identified. However, no association was found between UGT1A7 polymorphisms and the incidence of KRAS mutation.
These results suggest that there is a potential role of UGT1A7 polymorphisms as a potential risk factor for lung cancer. Further studies are warranted to find out specific
clinical characteristics and tumor biomarkers on these susceptible indivisuals. Moreover, it is also important to discover other risk factors that predispose patients to form lung cancer-related oncogenic mutations.
論文目次 誌謝 I
中文摘要 II
英文摘要 III
目次 IV
表目錄 VI
圖目錄 VIII
第1章 緒論 1
第2章 文獻探討 2
2.1 肺癌概述 2
2.1.1 肺癌簡介 2
2.1.2 肺癌之流行病學 4
2.1.3 肺癌致病危險因子 6
2.1.4 肺癌治療現況 8
2.2 肺癌致癌基因與生物標記 11
2.2.1 KRAS基因 11
2.2.2 EGFR基因 14
2.2.3 肺癌分子特徵之差異 15
2.3 代謝酶與肺癌 17
2.3.1 致癌物質與肺癌 17
2.3.2 代謝致癌物質的個體代謝差異與肺癌罹病率的關係 18
2.4 尿苷雙磷酸葡萄糖醛酸基轉移酶1A7(UDP -GLUCURONOSYLTRANSFERASE 1A7 ) 20
2.4.1 尿苷雙磷酸葡萄糖醛酸基轉移酶 (UGT) 簡介 20
2.4.2 尿苷雙磷酸葡萄糖醛酸基轉移酶1A7 (UGT1A7) 簡介 23
2.4.3 尿苷雙磷酸葡萄糖醛酸基轉移酶1A7的基因多型性 23
2.4.4 尿苷雙磷酸葡萄糖醛酸基轉移酶1A7與癌症的關係 27
第3章 研究目的 30
第4章 研究方法 31
4.1 研究設計 31
4.1.1 研究對象 31
4.1.2 資料收集 32
4.1.3 研究相關定義 32
4.2 UGT1A7基因型檢測方式 34
4.2.1 DNA萃取 (DNA Extraction) 35
4.2.2 聚合酶連鎖反應-限制酶切割片段長度多型性 (PCR-RFLP) 35
4.2.3 基因型判定 38
4.3 KRAS基因突變檢測方式 41
4.3.1 聚合酶連鎖反應-限制酶切割片段長度多型性 (PCR-RFLP) 42
4.3.2 KRAS基因型判定 47
4.4 統計分析 49
第5章 研究結果 50
5.1 病患組與對照組之結果比較 50
5.1.1 基本資料分析 50
5.1.2 病患組與對照組之UGT1A7對偶基因分析 52
5.1.3 病患組與對照組之UGT1A7基因型分析 53
5.1.4 不同性別研究對象之病患組與對照組UGT1A7基因型分析 54
5.1.5 不同病理型態病患組與對照組之UGT1A7基因型分析 56
5.1.6 不同吸菸習慣病患組與對照組之UGT1A7基因型分析 58
5.1.7 不同EGFR基因型態病患組與對照組之UGT1A7基因型分析 59
5.2 KRAS基因突變之結果比較 61
5.2.1 KRAS基因突變檢測結果 61
5.2.2 不同KRAS基因型態病人之比較結果 61
5.2.3 不同KRAS基因型態病人之UGT1A7基因型分析 63
5.2.4 KRAS基因型態與EGFR基因型態之關聯分析 64
5.2.5 KRAS基因與使用EGFR-TKI後腫瘤反應之關聯分析 64
第6章 討論 65
6.1 研究結果討論 65
6.1.1 UGT1A7對偶基因與基因型頻率分布比較 65
6.1.2 UGT1A7基因多型與肺癌之相關性 66
6.1.3 KRAS基因突變之結果討論 68
6.2 研究限制及未來改進方向 72
第7章 結論 73
參考文獻 74
參考文獻 1.行政院衛生署國民健康局. 中華民國95年癌症登記報告, 2009.

2.Hecht SS. Tobacco carcinogens, their biomarkers and tobacco-induced cancer. Nat Rev Cancer 2003; 3(10): 733-744.

3.Schubbert S, Shannon K, Bollag G. Hyperactive Ras in developmental disorders and cancer. Nat Rev Cancer 2007; 7(4): 295-308.

4.Guillemette C, Ritter JK, Auyeung DJ, Kessler FK, Housman DE. Structural heterogeneity at the UDP-glucuronosyltransferase 1 locus: functional consequences of three novel missense mutations in the human UGT1A7 gene. Pharmacogenetics 2000; 10(7): 629-644.

5.Nagar S, Remmel RP. Uridine diphosphoglucuronosyltransferase pharmacogenetics and cancer. Oncogene 2006; 25(11): 1659-1672.

6.Minna John D, Schiller Joan H, "Chapter 85. Neoplasms of the Lung" (Chapter). Fauci AS, Braunwald E, Kasper DL, Hauser SL, Longo DL, Jameson JL, Loscalzo J: Harrison's Principles of Internal Medicine, 17th Edition: http://www.accessmedicine.com/content.aspx?aID=2889473.

7.Collins LG, Haines C, Perkel R, Enck RE. Lung cancer: diagnosis and management. Am Fam Physician 2007; 75(1): 56-63.

8.Goldstraw P, Crowley J, Chansky K, Giroux DJ, Groome PA, Rami-Porta R, Postmus PE, Rusch V, Sobin L. The IASLC Lung Cancer Staging Project: proposals for the revision of the TNM stage groupings in the forthcoming (seventh) edition of the TNM Classification of malignant tumours. J Thorac Oncol 2007; 2(8): 706-714.

9.Shepherd FA, Crowley J, Van Houtte P, Postmus PE, Carney D, Chansky K, Shaikh Z, Goldstraw P. The International Association for the Study of Lung Cancer lung cancer staging project: proposals regarding the clinical staging of small cell lung cancer in the forthcoming (seventh) edition of the tumor, node, metastasis classification for lung cancer. J Thorac Oncol 2007; 2(12): 1067-1077.

10.Parkin DM, Bray F, Ferlay J, Pisani P. Global cancer statistics, 2002. CA Cancer J Clin 2005; 55(2): 74-108.

11.Liao KM, Chen SY, Chen CJ. Epidemiology of Lung Cancer: global and local characteristics [Lecture Article]. J Internal Med Taiwan 2002.

12.Alberg AJ, Ford JG, Samet JM. Epidemiology of lung cancer: ACCP evidence-based clinical practice guidelines (2nd edition). Chest 2007; 132(3 Suppl): 29S-55S.

13.Sun S, Schiller JH, Gazdar AF. Lung cancer in never smokers--a different disease. Nat Rev Cancer 2007; 7(10): 778-790.

14.Chen CL, Hsu LI, Chiou HY, Hsueh YM, Chen SY, Wu MM, Chen CJ. Ingested arsenic, cigarette smoking, and lung cancer risk: a follow-up study in arseniasis-endemic areas in Taiwan. JAMA 2004; 292(24): 2984-2990.

15.TCOG肺癌研究委員會. 肺癌臨床指引. 國家衛生研究院 癌症研究組 臺灣癌症臨床研究合作組織 2004.

16.Blackhall F, Ranson M, Thatcher N. Where next for gefitinib in patients with lung cancer? Lancet Oncol 2006; 7(6): 499-507.

17.Ciardiello F, Tortora G. EGFR antagonists in cancer treatment. N Engl J Med 2008; 358(11): 1160-1174.

18.Thatcher N, Chang A, Parikh P, Rodrigues Pereira J, Ciuleanu T, von Pawel J, Thongprasert S, Tan EH, Pemberton K, Archer V, Carroll K. Gefitinib plus best supportive care in previously treated patients with refractory advanced non-small-cell lung cancer: results from a randomised, placebo-controlled, multicentre study (Iressa Survival Evaluation in Lung Cancer). Lancet 2005; 366(9496): 1527-1537.

19.Shepherd FA, Rodrigues Pereira J, Ciuleanu T, Tan EH, Hirsh V, Thongprasert S, Campos D, Maoleekoonpiroj S, Smylie M, Martins R, van Kooten M, Dediu M, Findlay B, Tu D, Johnston D, Bezjak A, Clark G, Santabarbara P, Seymour L. Erlotinib in previously treated non-small-cell lung cancer. N Engl J Med 2005; 353(2): 123-132.

20.Hann CL, Brahmer JR. "Who should receive epidermal growth factor receptor inhibitors for non-small cell lung cancer and when?". Curr Treat Options Oncol 2007; 8(1): 28-37.

21.National Cancer Institute. Small Cell Lung Cancer Treatment (PDQ®) (Accessed 2009-05-10, at http://www.cancer.gov/cancertopics/pdq/treatment/small-cell-lung/HealthProfessional/page2#Section_156).

22.行政院衛生署國民健康局. 癌症期別五年存活率. (Accessed 2009-05-10, at http://crs.cph.ntu.edu.tw/uploadimages/04.%20Lung%20Cancer_ALCC.pdf).

23.Croce CM. Oncogenes and cancer. N Engl J Med 2008; 358(5): 502-511.

24Herbst RS, Heymach JV, Lippman SM. Lung cancer. N Engl J Med 2008; 359(13): 1367-1380.

25.DeMarini DM, Landi S, Tian D, Hanley NM, Li X, Hu F, Roop BC, Mass MJ, Keohavong P, Gao W, Olivier M, Hainaut P, Mumford JL. Lung tumor KRAS and TP53 mutations in nonsmokers reflect exposure to PAH-rich coal combustion emissions. Cancer Res 2001; 61(18): 6679-6681.

26.Akopyan G, Bonavida B. Understanding tobacco smoke carcinogen NNK and lung tumorigenesis. Int J Oncol 2006; 29(4): 745-752.

27.Wu CC, Hsu HY, Liu HP, Chang JW, Chen YT, Hsieh WY, Hsieh JJ, Hsieh MS, Chen YR, Huang SF. Reversed mutation rates of KRAS and EGFR genes in adenocarcinoma of the lung in Taiwan and their implications. Cancer 2008; 113(11): 3199-3208.

28.Pao W, Wang TY, Riely GJ, Miller VA, Pan Q, Ladanyi M, Zakowski MF, Heelan RT, Kris MG, Varmus HE. KRAS mutations and primary resistance of lung adenocarcinomas to gefitinib or erlotinib. PLoS Med 2005; 2(1): e17.

29.Linardou H, Dahabreh IJ, Kanaloupiti D, Siannis F, Bafaloukos D, Kosmidis P, Papadimitriou CA, Murray S. Assessment of somatic k-RAS mutations as a mechanism associated with resistance to EGFR-targeted agents: a systematic review and meta-analysis of studies in advanced non-small-cell lung cancer and metastatic colorectal cancer. Lancet Oncol 2008; 9(10): 962-972.

30.Slebos RJ, Kibbelaar RE, Dalesio O, Kooistra A, Stam J, Meijer CJ, Wagenaar SS, Vanderschueren RG, van Zandwijk N, Mooi WJ, et al. K-ras oncogene activation as a prognostic marker in adenocarcinoma of the lung. N Engl J Med 1990; 323(9): 561-565.

31.Zhu CQ, da Cunha Santos G, Ding K, Sakurada A, Cutz JC, Liu N, Zhang T, Marrano P, Whitehead M, Squire JA, Kamel-Reid S, Seymour L, Shepherd FA, Tsao MS. Role of KRAS and EGFR as biomarkers of response to erlotinib in National Cancer Institute of Canada Clinical Trials Group Study BR.21. J Clin Oncol 2008; 26(26): 4268-4275.

32.Karapetis CS, Khambata-Ford S, Jonker DJ, O'Callaghan CJ, Tu D, Tebbutt NC, Simes RJ, Chalchal H, Shapiro JD, Robitaille S, Price TJ, Shepherd L, Au HJ, Langer C, Moore MJ, Zalcberg JR. K-ras mutations and benefit from cetuximab in advanced colorectal cancer. N Engl J Med 2008; 359(17): 1757-1765.

33.Sharma SV, Bell DW, Settleman J, Haber DA. Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer 2007; 7(3): 169-181.

34.Eberhard DA, Giaccone G, Johnson BE. Biomarkers of response to epidermal growth factor receptor inhibitors in Non-Small-Cell Lung Cancer Working Group: standardization for use in the clinical trial setting. J Clin Oncol 2008; 26(6): 983-994.

35.Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW, Harris PL, Haserlat SM, Supko JG, Haluska FG, Louis DN, Christiani DC, Settleman J, Haber DA. Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med 2004; 350(21): 2129-2139.

36.Mitsudomi T, Yatabe Y. Mutations of the epidermal growth factor receptor gene and related genes as determinants of epidermal growth factor receptor tyrosine kinase inhibitors sensitivity in lung cancer. Cancer Sci 2007; 98(12): 1817-1824.

37.Pao W, Miller VA, Politi KA, Riely GJ, Somwar R, Zakowski MF, Kris MG, Varmus H. Acquired resistance of lung adenocarcinomas to gefitinib or erlotinib is associated with a second mutation in the EGFR kinase domain. PLoS Med 2005; 2(3): e73.

38.Subramanian J, Govindan R. Molecular genetics of lung cancer in people who have never smoked. Lancet Oncol 2008; 9(7): 676-682.

39.Wogan GN, Hecht SS, Felton JS, Conney AH, Loeb LA. Environmental and chemical carcinogenesis. Semin Cancer Biol 2004; 14(6): 473-486.

40.Gemignani F, Landi S, Szeszenia-Dabrowska N, Zaridze D, Lissowska J, Rudnai P, Fabianova E, Mates D, Foretova L, Janout V, Bencko V, Gaborieau V, Gioia-Patricola L, Bellini I, Barale R, Canzian F, Hall J, Boffetta P, Hung RJ, Brennan P. Development of lung cancer before the age of 50: the role of xenobiotic metabolizing genes. Carcinogenesis 2007; 28(6): 1287-1293.

41.Roth JA, Cox JD, Hong WK. Lung cancer, 3rd edn. Blackwell Pub.: Malden, Mass. ; Oxford, 2008.

42.Ye Z, Song H, Higgins JP, Pharoah P, Danesh J. Five glutathione s-transferase gene variants in 23,452 cases of lung cancer and 30,397 controls: meta-analysis of 130 studies. PLoS Med 2006; 3(4): e91.

43.蕭珮妤. 人類乙醯轉移酶基因型與腦瘤之關聯研究. 台北醫學大學, 2006.

44.Guillemette C. Pharmacogenomics of human UDP-glucuronosyltransferase enzymes. Pharmacogenomics J 2003; 3(3): 136-158.

45.Coffman BL, Rios GR, King CD, Tephly TR. Human UGT2B7 catalyzes morphine glucuronidation. Drug Metab Dispos 1997; 25(1): 1-4.

46.Formelli F, Barua AB, Olson JA. Bioactivities of N-(4-hydroxyphenyl) retinamide and retinoyl beta-glucuronide. FASEB J 1996; 10(9): 1014-1024.

47.張瑞家. 尿苷雙磷酸葡萄糖酫酸基轉移酶1A1(UGT1A1)基因多型與抗結核藥物治療期間肝損傷之關聯研究. 台北醫學大學, 2007.

48.Mackenzie PI, Bock KW, Burchell B, Guillemette C, Ikushiro S, Iyanagi T, Miners JO, Owens IS, Nebert DW. Nomenclature update for the mammalian UDP glycosyltransferase (UGT) gene superfamily. Pharmacogenet Genomics 2005; 15(10): 677-685.

49.Mackenzie PI, Owens IS, Burchell B, Bock KW, Bairoch A, Belanger A, Fournel-Gigleux S, Green M, Hum DW, Iyanagi T, Lancet D, Louisot P, Magdalou J, Chowdhury JR, Ritter JK, Schachter H, Tephly TR, Tipton KF, Nebert DW. The UDP glycosyltransferase gene superfamily: recommended nomenclature update based on evolutionary divergence. Pharmacogenetics 1997; 7(4): 255-269.

50.Gong QH, Cho JW, Huang T, Potter C, Gholami N, Basu NK, Kubota S, Carvalho S, Pennington MW, Owens IS, Popescu NC. Thirteen UDPglucuronosyltransferase genes are encoded at the human UGT1 gene complex locus. Pharmacogenetics 2001; 11(4): 357-368.

51.Tukey RH, Strassburg CP. Human UDP-glucuronosyltransferases: metabolism, expression, and disease. Annu Rev Pharmacol Toxicol 2000; 40: 581-616.

52.Gregory PA, Lewinsky RH, Gardner-Stephen DA, Mackenzie PI. Regulation of UDP glucuronosyltransferases in the gastrointestinal tract. Toxicol Appl Pharmacol 2004; 199(3): 354-363.

53.Zheng Z, Fang JL, Lazarus P. Glucuronidation: an important mechanism for detoxification of benzo[a]pyrene metabolites in aerodigestive tract tissues. Drug Metab Dispos 2002; 30(4): 397-403.

54.Bock KW, Raschko FT, Gschaidmeier H, Seidel A, Oesch F, Grove AD, Ritter JK. Mono- and Diglucuronide formation from benzo[a]pyrene and chrysene diphenols by AHH-1 cell-expressed UDP-glucuronosyltransferase UGT1A7. Biochem Pharmacol 1999; 57(6): 653-656.

55.UGT Nomenclature Commitee. June 2005. UGT Alleles Nomenclature Home Page. (Accessed 2009-05-10, at http://www.pharmacogenomics.pha.ulaval.ca/webdav/site/pharmacogenomics/shared/Nomenclature/UGT1A/UGT1A7.htm ).

56.Huang MJ, Yang SS, Lin MS, Huang CS. Polymorphisms of uridine-diphosphoglucuronosyltransferase 1A7 gene in Taiwan Chinese. World J Gastroenterol 2005; 11(6): 797-802.

57.Chen K, Jin M, Zhu Y, Jiang Q, Yu W, Ma X, Yao K. Genetic polymorphisms of the uridine diphosphate glucuronosyltransferase 1A7 and colorectal cancer risk in relation to cigarette smoking and alcohol drinking in a Chinese population. J Gastroenterol Hepatol 2006; 21(6): 1036-1041.

58.Araki J, Kobayashi Y, Iwasa M, Urawa N, Gabazza EC, Taguchi O, Kaito M, Adachi Y. Polymorphism of UDP-glucuronosyltransferase 1A7 gene: a possible new risk factor for lung cancer. Eur J Cancer 2005; 41(15): 2360-2365.

59.Strassburg CP, Vogel A, Kneip S, Tukey RH, Manns MP. Polymorphisms of the human UDP-glucuronosyltransferase (UGT) 1A7 gene in colorectal cancer. Gut 2002; 50(6): 851-856.

60.Zheng Z, Park JY, Guillemette C, Schantz SP, Lazarus P. Tobacco carcinogen-detoxifying enzyme UGT1A7 and its association with orolaryngeal cancer risk. J Natl Cancer Inst 2001; 93(18): 1411-1418.

61.Wang Y, Kato N, Hoshida Y, Otsuka M, Taniguchi H, Moriyama M, Shiina S, Kawabe T, Ito YM, Omata M. UDP-glucuronosyltransferase 1A7 genetic polymorphisms are associated with hepatocellular carcinoma in japanese patients with hepatitis C virus infection. Clin Cancer Res 2004; 10(7): 2441-2446.

62.Vogel A, Ockenga J, Ehmer U, Barut A, Kramer FJ, Tukey RH, Manns MP, Strassburg CP. Polymorphisms of the carcinogen detoxifying UDP-glucuronosyltransferase UGT1A7 in proximal digestive tract cancer. Z Gastroenterol 2002; 40(7): 497-502.

63.Vogel A, Kneip S, Barut A, Ehmer U, Tukey RH, Manns MP, Strassburg CP. Genetic link of hepatocellular carcinoma with polymorphisms of the UDP-glucuronosyltransferase UGT1A7 gene. Gastroenterology 2001; 121(5): 1136-1144.

64.Tseng CS, Tang KS, Lo HW, Ker CG, Teng HC, Huang CS. UDP-glucuronosyltransferase 1A7 genetic polymorphisms are associated with hepatocellular carcinoma risk and onset age. Am J Gastroenterol 2005; 100(8): 1758-1763.

65.Liu W, Innocenti F, Ratain MJ. Linkage disequilibrium across the UGT1A locus should not be ignored in association studies of cancer susceptibility. Clin Cancer Res 2005; 11(3): 1348-1349; author reply 1349.

66.van der Logt EM, Bergevoet SM, Roelofs HM, van Hooijdonk Z, te Morsche RH, Wobbes T, de Kok JB, Nagengast FM, Peters WH. Genetic polymorphisms in UDP-glucuronosyltransferases and glutathione S-transferases and colorectal cancer risk. Carcinogenesis 2004; 25(12): 2407-2415.

67.Tang KS, Chiu HF, Chen HH, Eng HL, Tsai CJ, Teng HC, Huang CS. Link between colorectal cancer and polymorphisms in the uridine-diphosphoglucuronosyltransferase 1A7 and 1A1 genes. World J Gastroenterol 2005; 11(21): 3250-3254.

68.Butler LM, Duguay Y, Millikan RC, Sinha R, Gagne JF, Sandler RS, Guillemette C. Joint effects between UDP-glucuronosyltransferase 1A7 genotype and dietary carcinogen exposure on risk of colon cancer. Cancer Epidemiol Biomarkers Prev 2005; 14(7): 1626-1632.

69.Ockenga J, Vogel A, Teich N, Keim V, Manns MP, Strassburg CP. UDP glucuronosyltransferase (UGT1A7) gene polymorphisms increase the risk of chronic pancreatitis and pancreatic cancer. Gastroenterology 2003; 124(7): 1802-1808.

70.Verlaan M, Drenth JP, Truninger K, Koudova M, Schulz HU, Bargetzi M, Kunzli B, Friess H, Cerny M, Kage A, Landt O, te Morsche RH, Rosendahl J, Luck W, Nickel R, Halangk J, Becker M, Macek M, Jr., Jansen JB, Witt H. Polymorphisms of UDP-glucuronosyltransferase 1A7 are not involved in pancreatic diseases. J Med Genet 2005; 42(10): e62.

71.te Morsche RH, Drenth JP, Truninger K, Schulz HU, Kage A, Landt O, Verlaan M, Rosendahl J, Macek M, Jr., Jansen JB, Witt H. UGT1A7 polymorphisms in chronic pancreatitis: an example of genotyping pitfalls. Pharmacogenomics J 2008; 8(1): 34-41.

72.Oken MM, Creech RH, Tormey DC, Horton J, Davis TE, McFadden ET, Carbone PP. Toxicity and response criteria of the Eastern Cooperative Oncology Group. Am J Clin Oncol 1982; 5(6): 649-655.

73.Therasse P, Arbuck SG, Eisenhauer EA, Wanders J, Kaplan RS, Rubinstein L, Verweij J, Van Glabbeke M, van Oosterom AT, Christian MC, Gwyther SG. New guidelines to evaluate the response to treatment in solid tumors. European Organization for Research and Treatment of Cancer, National Cancer Institute of the United States, National Cancer Institute of Canada. J Natl Cancer Inst 2000; 92(3): 205-216.

74.Matsuo K, Ito H, Yatabe Y, Hiraki A, Hirose K, Wakai K, Kosaka T, Suzuki T, Tajima K, Mitsudomi T. Risk factors differ for non-small-cell lung cancers with and without EGFR mutation: assessment of smoking and sex by a case-control study in Japanese. Cancer Sci 2007; 98(1): 96-101.

75.Lee CN, Yu MC, Bai KJ, Chang JH, Fang CL, Hsu HL, Huang BS, Lu PC, Liu HE. NAT2 fast acetylator genotypes are associated with an increased risk for lung cancer with wildtype epidermal growth factor receptors in Taiwan. Lung Cancer 2009; 64(1): 9-12.

76.Tam IY, Chung LP, Suen WS, Wang E, Wong MC, Ho KK, Lam WK, Chiu SW, Girard L, Minna JD, Gazdar AF, Wong MP. Distinct epidermal growth factor receptor and KRAS mutation patterns in non-small cell lung cancer patients with different tobacco exposure and clinicopathologic features. Clin Cancer Res 2006; 12(5): 1647-1653.

77.Bae NC, Chae MH, Lee MH, Kim KM, Lee EB, Kim CH, Park TI, Han SB, Jheon S, Jung TH, Park JY. EGFR, ERBB2, and KRAS mutations in Korean non-small cell lung cancer patients. Cancer Genet Cytogenet 2007; 173(2): 107-113.

78.Kosaka T, Yatabe Y, Endoh H, Kuwano H, Takahashi T, Mitsudomi T. Mutations of the epidermal growth factor receptor gene in lung cancer: biological and clinical implications. Cancer Res 2004; 64(24): 8919-8923.

79.Jassem J, Jassem E, Jakobkiewicz-Banecka J, Rzyman W, Badzio A, Dziadziuszko R, Kobierska-Gulida G, Szymanowska A, Skrzypski M, Zylicz M. P53 and K-ras mutations are frequent events in microscopically negative surgical margins from patients with nonsmall cell lung carcinoma. Cancer 2004; 100(9): 1951-1960.

80.Jimeno A, Messersmith WA, Hirsch FR, Franklin WA, Eckhardt SG. KRAS mutations and sensitivity to epidermal growth factor receptor inhibitors in colorectal cancer: practical application of patient selection. J Clin Oncol 2009; 27(7): 1130-1136.

81.Kosaka T, Yatabe Y, Onozato R, Kuwano H, Mitsudomi T. Prognostic implication of EGFR, KRAS, and TP53 gene mutations in a large cohort of Japanese patients with surgically treated lung adenocarcinoma. J Thorac Oncol 2009; 4(1): 22-29.

82.Riely GJ, Kris MG, Rosenbaum D, Marks J, Li A, Chitale DA, Nafa K, Riedel ER, Hsu M, Pao W, Miller VA, Ladanyi M. Frequency and distinctive spectrum of KRAS mutations in never smokers with lung adenocarcinoma. Clin Cancer Res 2008; 14(18): 5731-5734.

83.Wiener D, Fang JL, Dossett N, Lazarus P. Correlation between UDP-glucuronosyltransferase genotypes and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone glucuronidation phenotype in human liver microsomes. Cancer Res 2004; 64(3): 1190-1196.

84.Gallagher CJ, Muscat JE, Hicks AN, Zheng Y, Dyer AM, Chase GA, Richie J, Lazarus P. The UDP-glucuronosyltransferase 2B17 gene deletion polymorphism: sex-specific association with urinary 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol glucuronidation phenotype and risk for lung cancer. Cancer Epidemiol Biomarkers Prev 2007; 16(4): 823-828.

------------------------------------------------------------------------ 第 13 筆 ---------------------------------------------------------------------
系統識別號 U0007-2007200917303500
論文名稱(中文) 喜樹幼葉成分之研究
論文名稱(英文) Studies on the constituents of the young leaves of Camptotheca acuminata
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 97
學期 2
出版年 98
研究生(中文) 楊士平
學號 M303096004
學位類別 碩士
語文別 中文
口試日期 2009-06-19
論文頁數 206頁
口試委員 委員-郭悅雄
委員-吳天賞
指導教授-李慶國
關鍵字(中) 喜樹
紫樹科
喜樹鹼
生物活性導引分離
關鍵字(英) Camptotheca acuminata Decaisne
Nyssaceae
Camptothecin
Bioactivity-directed fractionation
學科別分類
中文摘要 喜樹 (Camptotheca acuminata Decaisne) 為紫樹科 (Nyssaceae) 喜樹屬植物,為原產於中國大陸的落葉性喬木,主要分布在四川、雲南跟廣西等溫暖潮濕地區。喜樹裡最著名的二次代謝物是為喜樹鹼(Camptothecin, CPT),在1960年代被發現具有良好的抗腫瘤活性。
本研究利用喜樹幼葉部以乙醇浸泡萃取,浸泡液經減壓濃縮後,濃縮液經由水、乙酸乙酯以及正丁醇作分配萃取。取乙酸乙酯層及正丁醇層部份分別利用矽膠管柱層析、全自動色層分析儀作初步分離,之後利用薄層層析法、高效能液相層析法等方法,分離純化得到41個化合物,其中包括九個aliphatic類、一個monoterpenoid類、一個diterpenoid類、二個triterpenoid類、一個steroid類、一個pheophorbide類、六個aromatic類、七個flavonoid類、一個norsesquiterpenoid類、一個aliphatic glycoside類、一個secoiridoid glycoside類、二個quinoline類、三個quinic acid類、四個others類,其中化合物CA36為新化合物。
英文摘要 Camptotheca acuminata Decaisne (Nyssaceae), a deciduous tree native to China, is widely distributed in relatively warm areas such as Sechwan, Yunnan, and Kwangsi. The secondary metabolites, Camptothecin (CPT) which was found to possess promising anti-tumor activity in the 1960s is isolated from C. acuminata.
In this study, the ethanol extract of young leaves of Camptotheca acuminata Decaisne was concentrated to give a residue which was partition with water, ethyl acetate, and butanol. The ethyl acetate layer and butanol layer were separated by column chromatography on silica gel, automated flash chromatography, thin layer chromatography, and high performance liquid chromatography to afford forty-one compounds. Including nine aliphatic compounds, one monoterpenoid compound, one diterpenoid compound, two triterpenoid compounds, one steroid compound, one pheophorbide compound, six aromatic compounds, seven flavonoid compounds, one norsesquiterpenoid compound, one aliphatic glycoside compound, one secoiridoid glycoside compound, two quinoline compounds, three quinic acid compounds, and four others compounds. Among them, CA36 is a new compound.
論文目次 謝誌---------------------------------------------I
中文摘要-----------------------------------------II
英文摘要-----------------------------------------III
目錄---------------------------------------------IV
表目錄-------------------------------------------VI
圖目錄-------------------------------------------VIII

第一章 緒論--------------------------------------1
第一節、研究動機及背景---------------------------1
第二節、植物分佈及形態---------------------------4
第三節、喜樹過去的成份研究概述-------------------6
第四節、喜樹過去之活性成分研究-------------------10
第五節、喜樹鹼及其衍生物的歷史回顧---------------12

第二章 實驗結果與討論----------------------------48
第一節、Loliolide之結構解析----------------------58
第二節、Syringaresinol之結構解析-----------------67
第三節、Kaempferol-3-β-D- (6-O-trans-p-coumaryl) glucopyranoside之結構解析------------------------74
第四節、Icariside B1之結構解析-------------------79
第五節、(Z)-3-hexenyl β-D-glucopyranoside之結構解析-----87
第六節、Vogeloside之結構解析--------------------92
第七節、Camptothecin之結構解析------------------101
第八節、10-methoxycamptothecin之結構解析--------106
第九節、3-O-trans-coumaryolquinic acid methyl ester之結構解析----------------------------------------------112
第十節、3-O-cis-coumaryolquinic acid methyl ester之結構解析----------------------------------------------119
第十一節、3-O-caffeoylquinic acid methyl ester之結構解析---------------------------------------------------126
第十二節、α-tocospiro A之結構解析---------------131
第十三節、α-tocospiro B之結構解析---------------139

第三章 生物活性試驗-----------------------------145
第一節、活性試驗步驟----------------------------145
第二節、活性試驗結果----------------------------148

第四章 實驗方法、材料及數據整理-----------------150
第一節、實驗儀器及藥品--------------------------150
第二節、植物材料與成分分離流程細節--------------153
第三節、各化合物之光譜數據總整理----------------160

第五章 結論-------------------------------------185

參考文獻----------------------------------------187
參考文獻 1. Qin, H.; Phengklai, C., Nyssaceae in Flora of China. Flora of China 2007, 13, 300-303.
2. 甘偉松等, 中國高等植物圖鑑 (二). 宏業書局: 1983; p 985.
3. 邱年永; 張光雄, 原色台灣藥用植物圖鑑 (5). 南天書局: 1998; Vol. 5, p 148.
4. Wall, M. E.; Wani, M. C.; Cook, C. E.; Palmer, K. H.; McPhail, A. T.; Sim, G. A., Plant Antitumor Agents. I. The Isolation and Structure of Camptothecin, a Novel Alkaloidal Leukemia and Tumor Inhibitor from Camptotheca acuminata. Journal of the American Chemical Society 1966, 88, (16), 3888-3890.
5. Hsiang, Y. H.; Hertzberg, R.; Hecht, S.; Liu, L. F., Camptothecin induces protein-linked DNA breaks via mammalian DNA topoisomerase I. The Journal of biological chemistry 1985, 260, (27), 14873-8.
6. Zhang, Z.; Li, S.; Zhang, S.; Liang, C.; Gorenstein, D.; Beasley, R. S., New Camptothecin and Ellagic Acid Analogues from the Root Bark of Camptotheca acuminata. Planta Medica 2004, 70, (12), 1216-1221.
7. Wu, T. S.; Chen, M. T.; Kuoh, C. S.; Tien, H. J.; Yeh, M. Y., Studies on the Constituents of Camptotheca Acuminata Decne, Ⅱ. The Constituents of Fresh Fruit. Journal of the Chinese Chemical Society 1985, 32, (2), 173-175.
8. Adamovics, J. A.; Cina, J. A.; Richard Hutchinson, C., Minor alkaloids of Camptotheca acuminata. Phytochemistry 1979, 18, (6), 1085-1086.
9. Wani, M. C.; Wall, M. E., Plant antitumor agents. II. Structure of two new alkaloids from Camptotheca acuminata. The Journal of Organic Chemistry 1969, 34, (5), 1364-1367.
10. Wall, M. E.; Wani, M. C.; Natschke, S. M.; Nicholas, A. W., Plant antitumor agents. 22. Isolation of 11-hydroxycamptothecin from Camptotheca acuminata decne: total synthesis and biological activity. Journal of Medicinal Chemistry 1986, 29, (8), 1553-1555.
11. Lin, L. Z.; Cordell, G. A., Quinoline alkaloids from Camptotheca acuminata. Phytochemistry 1989, 28, (4), 1295-1297.
12. Carte, B. K.; DeBrosse, C.; Eggleston, D.; Hemling, M.; Mentzer, M.; Poehland, B.; Troupe, N.; Westley, J. W.; Hecht, S. M., Isolation and characterization of a presumed biosynthetic precursor of camptothecin from extracts of Camptotheca acuminata. Tetrahedron 1990, 46, (8), 2747-2760.
13. Lin, L. Z.; Cordell, G. A., 19-O-Methylangustoline from Camptotheca acuminata. Phytochemistry 1990, 29, (8), 2744-2746.
14. Tan, Q., Yan, X.; Lin X.; Huang, Y.; Zheng, Z.; Song, S.; Lu, C.; Shen, Y., Chemical Constituents of the Endophytic Fungal Strain Phomopsis sp. NXZ-05 of Camptotheca acuminata. Helvetica Chimica Acta 2007, 90, (9), 1811-1817.
15. Hatano, T.; Ikegami, Y.; Shingu, T.; Okuda, T., Camptothins A and B, New Dimeric Hydrolyzable Tannins from Camptotheca acuminata DECNE. Chemical & Pharmaceutical Bulletin 1988, 36, (6), 2017-2022.
16. Bedir, E.; Tatli, I. I.; Khan, R. A.; Zhao, J.; Takamatsu, S.; Walker, L. A.; Goldman, P.; Khan, I. A., Biologically Active Secondary Metabolites from Ginkgo biloba. Journal of Agricultural and Food Chemistry 2002, 50, (11), 3150-3155.
17. Wei, F.; Ma, S. C.; Ma, L. Y.; But, P. P. H.; Lin, R. C.; Khan, I. A., Antiviral Flavonoids from the Seeds of Aesculus chinensis. Journal of Natural Products 2004, 67, (4), 650-653.
18. Mitrocotsa, D.; Mitaku, S.; Axarlis, S.; Harvala, C.; Malamas, M., Evaluation of the Antiviral Activity of Kaempferol and its Glycosides Against Human Cytomegalovirus. Planta Medica 2000, 66, (04), 377-379.
19. Inaba, H.; Tagashira, M.; Honma, D.; Kanda, T.; Kou, Y.; Ohtake, Y.; Amano, A., Identification of Hop Polyphenolic Components Which Inhibit Prostaglandin E2 Production by Gingival Epithelial Cells Stimulated with Periodontal Pathogen. Biological & Pharmaceutical Bulletin 2008, 31, (3), 527-530.
20. Masuda, T.; Iritani, K.; Yonemori, S.; Oyama, Y.; Takeda, Y., Isolation and Antioxidant Activity of Galloyl Flavonol Glycosides from the Seashore Plant, Pemphis acidula. Bioscience, Biotechnology, and Biochemistry 2001, 65, (6), 1302-1309.
21. Leung, H. W. C.; Lin, C. J.; Hour, M. J.; Yang, W. H.; Wang, M. Y.; Lee, H. Z., Kaempferol induces apoptosis in human lung non-small carcinoma cells accompanied by an induction of antioxidant enzymes. Food and Chemical Toxicology 2007, 45, (10), 2005-2013.
22. Park, K. Y.; Lee, S. H.; Min, B. K.; Lee, K. S.; Choi, J. S.; Chung, S. R.; Min, K. R.; Kim, Y., Inhibitory Effect of Luteolin 4'-O-Glucoside from Kummerowia striata and Other Flavonoids on Interleukin-5 Bioactivity. Planta Medica 1999, 65, (05), 457-459.
23. Wang, L.; Tu, Y. C.; Lian, T. W.; Hung, J. T.; Yen, J. H.; Wu, M. J., Distinctive Antioxidant and Antiinflammatory Effects of Flavonols. Journal of Agricultural and Food Chemistry 2006, 54, (26), 9798-9804.
24. Ramiro, E.; Franch, A.; Castellote, C.; Perez-Cano, F.; Permanyer, J.; Izquierdo-Pulido, M.; Castell, M., Flavonoids from Theobroma cacao Down-Regulate Inflammatory Mediators. Journal of Agricultural and Food Chemistry 2005, 53, (22), 8506-8511.
25. van der Sluis, A. A.; Dekker, M.; Verkerk, R.; Jongen, W. M. F., An Improved, Rapid in Vitro Method To Measure Antioxidant Activity. Application on Selected Flavonoids and Apple Juice. Journal of Agricultural and Food Chemistry 2000, 48, (9), 4116-4122.
26. Hammer, K. D. P.; Hillwig, M. L.; Solco, A. K. S.; Dixon, P. M.; Delate, K.; Murphy, P. A.; Wurtele, E. S.; Birt, D. F., Inhibition of Prostaglandin E2 Production by Anti-inflammatory Hypericum perforatum Extracts and Constituents in RAW264.7 Mouse Macrophage Cells. Journal of Agricultural and Food Chemistry 2007, 55, (18), 7323-7331.
27. Chow, J. M.; Shen, S. C.; Huan, S. K.; Lin, H. Y.; Chen, Y. C., Quercetin, but not rutin and quercitrin, prevention of H2O2-induced apoptosis via anti-oxidant activity and heme oxygenase 1 gene expression in macrophages. Biochemical Pharmacology 2005, 69, (12), 1839-1851.
28. Gatto, M. T.; Falcocchio, S.; Grippa, E.; Mazzanti, G.; Battinelli, L.; Nicolosi, G.; Lambusta, D.; Saso, L., Antimicrobial and Anti-Lipase Activity of Quercetin and its C2-C16 3-O-Acyl-Esters. Bioorganic & Medicinal Chemistry 2002, 10, (2), 269-272.
29. Wani, M. C.; Nicholas, A. W.; Wall, M. E., Plant antitumor agents. 28. Resolution of a key tricyclic synthon, 5'(RS)-1,5-dioxo-5'-hydroxy-2'H,5'H,6'H-6'-oxopyrano[3',4'-f].DELTA.6,8-tetrahydroindolizine: total synthesis and antitumor activity of 20(S)- and 20(R)-camptothecin. Journal of Medicinal Chemistry 1987, 30, (12), 2317-2319.
30. Jew, S. S.; Kim, H. J.; Kim, M. G.; Roh, E. Y.; Cho, Y. S.; Kim, J. K.; Cha, K. H.; Lee, K. K.; Han, H. J.; Choi, J. Y.; Lee, H., Synthesis and antitumor activity of 7-substituted 20(RS)-camptothecin analogues. Bioorganic & Medicinal Chemistry Letters 1996, 6, (7), 845-848.
31. Del Poeta, M.; Chen, S. F.; Von Hoff, D.; Dykstra, C. C.; Wani, M. C.; Manikumar, G.; Heitman, J.; Wall, M. E.; Perfect, J. R., Comparison of In Vitro Activities of Camptothecin and Nitidine Derivatives against Fungal and Cancer Cells. Antimicrobial Agents and Chemotherapy 1999, 43, (12), 2862-2868.
32. Werbovetz, K. A.; Bhattacharjee, A. K.; Brendle, J. J.; Scovill, J. P., Analysis of stereoelectronic properties of camptothecin analogues in relation to biological activity. Bioorganic & Medicinal Chemistry 2000, 8, (7), 1741-1747.
33. Bodley, A. L.; Cumming, J. N.; Shapiro, T. A., Effects of camptothecin, a topoisomerase I inhibitor, on Plasmodium falciparum. Biochemical Pharmacology 1998, 55, (5), 709-711.
34. Hertzberg, R. P.; Caranfa, M. J.; Holden, K. G.; Jakas, D. R.; Gallagher, G.; Mattern, M. R.; Mong, S. M.; Bartus, J. O. L.; Johnson, R. K.; Kingsbury, W. D., Modification of the hydroxylactone ring of camptothecin: inhibition of mammalian topoisomerase I and biological activity. Journal of Medicinal Chemistry 1989, 32, (3), 715-720.
35. Wani, M. C.; Nicholas, A. W.; Wall, M. E., Plant antitumor agents. 23. Synthesis and antileukemic activity of camptothecin analogs. Journal of Medicinal Chemistry 1986, 29, (11), 2358-2363.
36. Wani, M. C.; Ronman, P. E.; Lindley, J. T.; Wall, M. E., Plant antitumor agents. 18. Synthesis and biological activity of camptothecin analogs. Journal of Medicinal Chemistry 1980, 23, (5), 554-560.
37. Xiao, X.; Antony, S.; Pommier, Y.; Cushman, M., Total Synthesis and Biological Evaluation of 22-Hydroxyacuminatine. Journal of Medicinal Chemistry 2006, 49, (4), 1408-1412.
38. Sun, J.; Lou, H.; Dai, S.; Xu, H.; Zhao, F.; Liu, K., Indole alkoloids from Nauclea officinalis with weak antimalarial activity. Phytochemistry 2008, 69, (6), 1405-1410.
39. Wang, R. F.; Xie, W. D.; Zhang; Xing, D. M.; Ding, Y.; Wang, W.; Ma, C.; Du, L. J., Bioactive Compounds from the Seeds of Punica granatum (Pomegranate). Journal of Natural Products 2004, 67, (12), 2096-2098.
40. Deng, J. Z.; Marshall, R.; Jones, S. H.; Johnson, R. K.; Hecht, S. M., DNA-Damaging Agents from Crypteronia paniculata. Journal of Natural Products 2002, 65, (12), 1930-1932.
41. Miyamoto, K.; Kishi, N.; Koshiura, R.; Yoshida, T.; Hatano, T.; Okuda, T., Relationship between the Structures and the Antitumor Activities of Tannins. Chemical & Pharmaceutical Bulletin 1987, 35, (2), 814-822.
42. Fukuda, T.; Ito, H.; Yoshida, T., Antioxidative polyphenols from walnuts (Juglans regia L.). Phytochemistry 2003, 63, (7), 795-801.
43. Kim, H. J.; Lee, J. S.; Woo, E. R.; Kim, M. K.; Yang, B. S.; Yu, Y. G.; Hokoon, Y.; Park; Lee, S., Isolation of Virus-Cell Fusion Inhibitory Components from Eugenia caryophyllata. Planta Medica 2001, 67, (03), 277-279.
44. Kolodziej, H.; Kayser, O.; Kiderlen, A. F.; Ito, H.; Hatano, T.; Yoshida, T.; Foo, L. Y., Antileishmanial Activity of Hydrolyzable Tannins and their Modulatory Effects on Nitric Oxide and Tumour Necrosis Factor-α Release in Macrophages in Vitro. Planta Medica 2001, 67, (09), 825-832.
45. Khennouf, S.; Benabdallah, H.; Gharzouli, K.; Amira, S.; Ito, H.; Kim, T. H.; Yoshida, T.; Gharzouli, A., Effect of Tannins from Quercus suber and Quercus coccifera Leaves on Ethanol-Induced Gastric Lesions in Mice. Journal of Agricultural and Food Chemistry 2003, 51, (5), 1469-1473.
46. Etzenhouser, B.; Hansch, C.; Kapur, S.; Selassie, C. D., Mechanism of toxicity of esters of Caffeic and dihydrocaffeic acids. Bioorganic & Medicinal Chemistry 2001, 9, (1), 199-209.
47. Nagaoka, T.; Banskota, A. H.; Tezuka, Y.; Midorikawa, K.; Matsushige, K.; Kadota, S., Caffeic Acid Phenethyl Ester (CAPE) Analogues: Potent Nitric Oxide Inhibitors from the Netherlands Propolis. Biological & Pharmaceutical Bulletin 2003, 26, (4), 487-491.
48. Ma, C. M.; Cai, S. Q.; Cui, J. R.; Wang, R. Q.; Tu, P. F.; Hattori, M.; Daneshtalab, M., The cytotoxic activity of ursolic acid derivatives. European Journal of Medicinal Chemistry 2005, 40, (6), 582-589.
49. Banno, N.; Akihisa, T.; Tokuda, H.; Yasukawa, K.; Taguchi, Y.; Akazawa, H.; Ukiya, M.; Kimura, Y.; Suzuki, T.; Nishino, H., Anti-inflammatory and Antitumor-Promoting Effects of the Triterpene Acids from the Leaves of Eriobotrya japonica. Biological & Pharmaceutical Bulletin 2005, 28, (10), 1995-1999.
50. V. Leite, J. P.; Oliveira, A. B.; Lombardi, J. A.; Filho, J. D. S.; Chiari, E., Trypanocidal Activity of Triterpenes from Arrabidaea triplinervia and Derivatives. Biological & Pharmaceutical Bulletin 2006, 29, (11), 2307-2309.
51. Urban, M.; Sarek, J.; Klinot, J.; Korinkova, G.; Hajduch, M., Synthesis of A-Seco Derivatives of Betulinic Acid with Cytotoxic Activity. Journal of Natural Products 2004, 67, (7), 1100-1105.
52. Ziegler, H. L.; Franzyk, H.; Sairafianpour, M.; Tabatabai, M.; Tehrani, M. D.; Bagherzadeh, K.; Hägerstrand, H.; Stærk, D.; Jaroszewski, J. W., Erythrocyte membrane modifying agents and the inhibition of Plasmodium falciparum growth: structure-activity relationships for betulinic acid analogues. Bioorganic & Medicinal Chemistry 2004, 12, (1), 119-127.
53. Chang, K. C.; Duh, C. Y.; Chen, I. S.; Tsai, I. L., A Cytotoxic Butenolide, Two New Dolabellane Diterpenoids, a Chroman and a Benzoquinol Derivative Formosan Casearia membranacea. Planta Medica 2003, 69, (07), 667-672.
54. Jin, J. L.; Lee, S.; Lee, Y. Y.; Kim, J. M.; Heo, J. E.; Yun-Choi, H. S., Platelet Anti-Aggregating Triterpenoids from the Leaves of Acanthopanax senticosus and the Fruits of A. sessiliflorus. Planta Medica 2004, 70, (06), 564-566.
55. Kim, S. H.; Shin, D.S.; Oh, M. N.; Chung, S. C.; Lee, J. S.; Chang, I.-M.; Oh, K. B., Inhibition of Sortase, a Bacterial Surface Protein Anchoring Transpeptidase, by β-Sitosterol-3-O-glucopyranoside from Fritillaria verticillata. Bioscience, Biotechnology, and Biochemistry 2003, 67, (11), 2477-2479.
56. Kuete V.; Eyong, K. O.; Folefoc, G. N.; Beng, V. P.; Hussain, H.; Krohn, K.; Nkengfack, A. E., Antimicrobial activity of the methanolic extract and of the chemical constituents isolated from Newbouldia laevis. Pharmazie 2007, 62, (7), 552-556.
57. Oberlies, N. H.; Kroll, D. J., Camptothecin and taxol: historic achievements in natural products research. Journal of Natural Products 2004, 67, (2), 129-35.
58. Wall, M. E., Camptothecin and Taxol : Discovery to Clinic. Medicinal Research Reviews 1998, 18, (5), 299-314.
59. Gottlieb, J. A.; Luce, J. K., Treatment of malignant melanoma with camptothecin (NSC-100880). Cancer chemotherapy reports 1972, 56, (1), 103-5.
60. Moertel, C. G.; Schutt, A. J.; Reitemeier, R. J.; Hahn, R. G., Phase II study of camptothecin (NSC-100880) in the treatment of advanced gastrointestinal cancer. Cancer chemotherapy reports 1972, 56, (1), 95-101.
61. Hsiang, Y. H.; Lihou, M. G.; Liu, L. F., Arrest of replication forks by drug-stabilized topoisomerase I-DNA cleavable complexes as a mechanism of cell killing by camptothecin. Cancer Research 1989, 49, (18), 5077-5082.
62. Chen, A. Y.; Liu, L. F., DNA topoisomerases: essential enzymes and lethal targets. Annual Review of Pharmacology and Toxicology 1994, 34, 191-218.
63. Wall, M. E.; Wani, M. C.; Nicholas, A. W.; Manikumar, G.; Tele, C.; Moore, L.; Truesdale, A.; Leitner, P.; Besterman, J. M., Plant antitumor agents. 30. Synthesis and structure activity of novel camptothecin analogs. Journal of Medicinal Chemistry 1993, 36, (18), 2689-2700.
64. Wani, M. C.; Nicholas, A. W.; Manikumar, G.; Wall, M. E., Plant antitumor agents. 25. Total synthesis and antileukemic activity of ring A substituted camptothecin analogs. Structure-activity correlations. Journal of Medicinal Chemistry 1987, 30, (10), 1774-1779.
65. Crow, R. T.; Crothers, D. M., Structural modifications of camptothecin and effects on topoisomerase I inhibition. Journal of Medicinal Chemistry 1992, 35, (22), 4160-4164.
66. Kawato, Y.; Aonuma, M.; Hirota, Y.; Kuga, H.; Sato, K., Intracellular roles of SN-38, a metabolite of the camptothecin derivative CPT-11, in the antitumor effect of CPT-11. Cancer Research 1991, 51, (16), 4187-4191.
67. Raymond, E.; Campone, M.; Stupp, R.; Menten, J.; Chollet, P.; Lesimple, T.; Fety-Deporte, R.; Lacombe, D.; Paoletti, X.; Fumoleau, P., Multicentre phase II and pharmacokinetic study of RFS2000 (9-nitro-camptothecin) administered orally 5 days a week in patients with glioblastoma multiforme. European Journal of Cancer 2002, 38, (10), 1348-1350.
68. Schöffski, P.; Herr, A.; Vermorken, J. B.; Van den Brande, J.; Beijnen, J. H.; Rosing, H.; Volk, J.; Ganser, A.; Adank, S.; Botma, H. J.; Wanders, J., Clinical phase II study and pharmacological evaluation of rubitecan in non-pretreated patients with metastatic colorectal cancer--significant effect of food intake on the bioavailability of the oral camptothecin analogue. European Journal of Cancer 2002, 38, (6), 807-813.
69. Josien, H.; Bom, D.; Curran, D. P.; Zheng, Y. H.; Chou, T. C., 7-Silylcamptothecins (silatecans): A new family of camptothecin antitumor agents. Bioorganic & Medicinal Chemistry Letters 1997, 7, (24), 3189-3194.
70. Bom, D.; Curran, D. P.; Chavan, A. J.; Kruszewski, S.; Zimmer, S. G.; Fraley, K. A.; Burke, T. G., Novel A,B,E-Ring-Modified Camptothecins Displaying High Lipophilicity and Markedly Improved Human Blood Stabilities. Journal of Medicinal Chemistry 1999, 42, (16), 3018-3022.
71. Bom, D.; Curran, D. P.; Kruszewski, S.; Zimmer, S. G.; Thompson Strode, J.; Kohlhagen, G.; Du, W.; Chavan, A. J.; Fraley, K. A.; Bingcang, A. L.; Latus, L. J.; Pommier, Y.; Burke, T. G., The Novel Silatecan 7-tert-Butyldimethylsilyl-10-hydroxycamptothecin Displays High Lipophilicity, Improved Human Blood Stability, and Potent Anticancer Activity. Journal of Medicinal Chemistry 2000, 43, (21), 3970-3980.
72. Jaxel, C.; Kohn, K. W.; Wani, M. C.; Wall, M. E.; Pommier, Y., Structure-activity study of the actions of camptothecin derivatives on mammalian topoisomerase I: evidence for a specific receptor site and a relation to antitumor activity. Cancer Research 1989, 49, (6), 1465-1469.
73. Hsiang, Y. H.; Liu, L. F.; Wall, M. E.; Wani, M. C.; Nicholas, A. W.; Manikumar, G.; Kirschenbaum, S.; Silber, R.; Potmesil, M., DNA topoisomerase I-mediated DNA cleavage and cytotoxicity of camptothecin analogues. Cancer Research 1989, 49, (16), 4385-4389.
74. Fan, Y.; Weinstein, J. N.; Kohn, K. W.; Shi, L. M.; Pommier, Y., Molecular modeling studies of the DNA-topoisomerase I ternary cleavable complex with camptothecin. Journal of Medicinal Chemistry 1998, 41, (13), 2216-2226.
75. Mitsui, I.; Kumazawa, E.; Hirota, Y.; Aonuma, M.; Sugimori, M.; Ohsuki, S.; Uoto, K.; Ejima, A.; Terasawa, H.; Sato, K., A New Water-soluble Camptothecin Derivative, DX-8951f, Exhibits Potent Antitumor Activity against Human Tumors in vitro and in vivo. Japanese Journal of Cancer Research 1995, 86, (8), 776-782.
76. van Hattum, A. H.; Pinedo, H. M.; Schlüper, H. M. M.; Erkelens, C. A. M.; Tohgo, A.; Boven, E., The activity profile of the hexacyclic camptothecin derivative DX-8951f in experimental human colon cancer and ovarian cancer. Biochemical Pharmacology 2002, 64, (8), 1267-1277.
77. Ihara, M.; Noguchi, K.; Ohsawa, T.; Fukumoto, K.; Kametani, T., Studies on the syntheses of heterocyclic compounds and natural products. 999. Double enamine annelation of 3,4-dihydro-1-methyl-.beta.-carboline and isoquinoline derivatives with 6-methyl-2-pyrone-3,5-dicarboxylates and its application for the synthesis of (.+-.)-camptothecin. The Journal of Organic Chemistry 1983, 48, (19), 3150-3156.
78. Kurihara, T.; Tanno, H.; Takemura, S.; Harusawa, S.; Yoneda, R., Synthesis of C-nor-4, 6-Secocamptothecin and Related Compound. Journal of Heterocyclic Chemistry 1993, 30, 643-652.
79. Nicholas, A. W.; Wani, M. C.; Manikumar, G.; Wall, M. E.; Kohn, K. W.; Pommier, Y., Plant antitumor agents. 29. Synthesis and biological activity of ring D and ring E modified analogs of camptothecin. Journal of Medicinal Chemistry 1990, 33, (3), 972-978.
80. Sawada, S.; Nokata, K.; Furuta, T.; Yokokura, T.; Miyasaka, T., Chemical modification of an antitumor alkaloid camptothecin: synthesis and antitumor activity of 7-C-substituted camptothecins. Chemical & Pharmaceutical Bulletin 1991, 39, (10), 2574-2580.
81. Subrahmanyam, D.; Venkateswarlu, A.; Rao, K. V.; Sastry, T. V. R. S.; Vandana, G.; Kumar, S. A., Novel C-ring analogues of 20(S)-camptothecin-part-2: Synthesis and cytotoxicity of 5-C-substituted 20(S)-camptothecin analogues. Bioorganic & Medicinal Chemistry Letters 1999, 9, (12), 1633-1638.
82. Sugimori, M.; Ejima, A.; Ohsuki, S.; Matsumoto, K.; Kawato, Y.; Yasuoka, M.; Tagawa, H.; Terasawa, H., Antitumor Agents. VI. Synthesis and Antitumor Activity of Ring A-, Ring B-, and Ring C-Modified Derivatives of Camptothecin. Heterocycles 1994, 38, (1), 81-94.
83. Fassberg, J.; Stella, V. J., A kinetic and mechanistic study of the hydrolysis of camptothecin and some analogues. Journal of Pharmaceutical Sciences 1992, 81, (7), 676-684.
84. Ejima, A.; Terasawa, H.; Sugimori, M.; Ohsuki, S.; Matsumoto, K.; Kawato, Y.; Yasuoka, M.; Tagawa, H., Antitumor agents. V. Synthesis and antileukemic activity of E-ring-modified (RS)-camptothecin analogues. Chemical & Pharmaceutical Bulletin 1992, 40, (3), 683-688.
85. Wang, X.; Zhou, X.; Hecht, S. M., Role of the 20-Hydroxyl Group in Camptothecin Binding by the Topoisomerase I-DNA Binary Complex. Biochemistry 1999, 38, (14), 4374-4381.
86. Zunino, F.; Dallavalleb, S.; Laccabuea, D.; Berettaa, G.; Merlinib, L.; Pratesi, G., Current status and perspectives in the development of camptothecins. Current pharmaceutical design 2002, 8, (27), 2505-2520.
87. Lavergne, O.; Lesueur-Ginot, L.; Rodas, F. P.; Bigg, D. C. H., BN 80245: An E-ring modified camptothecin with potent antiproliferative and topoisomerase I inhibitory activities. Bioorganic & Medicinal Chemistry Letters 1997, 7, (17), 2235-2238.
88. Lavergne, O.; Lesueur-Ginot, L.; Pla Rodas, F.; Kasprzyk, P. G.; Pommier, J.; Demarquay, D.; Prevost, G.; Ulibarri, G.; Rolland, A.; Schiano-Liberatore, A. M.; Harnett, J.; Pons, D.; Camara, J.; Bigg, D. C., Homocamptothecins: synthesis and antitumor activity of novel E-ring-modified camptothecin analogues. Journal of Medicinal Chemistry 1998, 41, (27), 5410-5419.
89. Lansiaux, A.; Facompre, M.; Wattez, N.; Hildebrand, M. P.; Bal, C.; Demarquay, D.; Lavergne, O.; Bigg, D. C. H.; Bailly, C., Apoptosis Induced by the Homocamptothecin Anticancer Drug BN80915 in HL-60 Cells. Molecular pharmacology 2001, 60, (3), 450-461.
90. Larsen, A. K.; Gilbert, C.; Chyzak, G.; Plisov, S. Y.; Naguibneva, I.; Lavergne, O.; Lesueur-Ginot, L.; Bigg, D. C. H., Unusual Potency of BN 80915, a Novel Fluorinated E-ring Modified Camptothecin, toward Human Colon Carcinoma Cells. Cancer Research 2001, 61, (7), 2961-2967.
91. Walker, M. A.; Dubowchik, G. M.; Hofstead, S. J.; Trail, P. A.; Firestone, R. A., Synthesis of an immunoconjugate of camptothecin. Bioorganic & Medicinal Chemistry Letters 2002, 12, (2), 217-219.
92. Aiyama, R.; Nagai, H.; Nokata, K.; Shinohara, C.; Sawada, S., A camptothecin derivative from nothapodytes foetida. Phytochemistry 1988, 27, (11), 3663-3664.
93. Saito, K.; Sudo, H.; Yamazaki, M.; Koseki-Nakamura, M.; Kitajima, M.; Takayama, H.; Aimi, N., Feasible production of camptothecin by hairy root culture of Ophiorrhiza pumila. Plant Cell Reports 2001, 20, (3), 267-271.
94. Gunasekera, S. P.; Badawi, M. M.; Cordell, G. A.; Farnsworth, N. R.; Chitnis, M., Plant anticancer agents X. Isolation of camptothecin and 9-methoxycamptothecin from Ervatamia heyneana. Journal of Natural Products 1979, 42, (5), 475-477.
95. López-Meyer, M.; Nessler, C. L.; McKnight, T. D., Sites of accumulation of the antitumor alkaloid camptothecin in Camptotheca acuminata. Planta Medica 1994, 60, (6), 558-560.
96. Li, S.; Yi, Y.; Wang, Y.; Zhang, Z.; Beasley, R. S., Camptothecin accumulation and variations in camptotheca. Planta Medica 2002, 68, (11), 1010-1016.
97. Lorence, A.; Medina-Bolivar, F.; Nessler, C. L., Camptothecin and 10-hydroxycamptothecin from Camptotheca acuminata hairy roots. Plant Cell Reports 2004, 22, (6), 437-441.
98. Wiedenfeld, H.; Furmanowa, M.; Roeder, E.; Guzewska, J.; GustowGustowski, W., Camptothecin and 10-hydroxycamptothecin in callus and plantlets of Camptotheca acuminata. Plant Cell, Tissue Organ Culture 1997, 49, 213-218.
99. Sakato, K.; Tanaka, H.; Mukai, N.; Misawa, M., Isolation and identification of camptothecin from cells of Camptotheca acuminata suspension cultures. Agricultural and Biological Chemistry 1974, 38, 217-218.
100. van Hengel, A. J.; Harkes, M. P.; Wichers, H. J.; Hesselink, P. G. M.; Buitelaar, R. M., Characterization of callus formation and camptothecin production by cell lines of Camptotheca acuminata. Plant Cell Tiss. Org. Cult. 1992, 28, 11-18.
101. Wu, T. S.; Leu, Y. L.; Hsu, H. C.; Ou, L. F.; Chen, C. C.; Chen, C. F.; Ou, J. C.; Wu, Y. C., Constituents and cytotoxic principles of Nothapodytes foetida. Phytochemistry 1995, 39, (2), 383-385.
102. Roja, G.; Heble, M. R., The quinoline alkaloids camptothecin and 9-methoxycamptothecin from tissue cultures and mature trees of Nothapodytes foetida. Phytochemistry 1994, 36, (1), 65-66.
103. Wu, S. F.; Hsieh, P. W.; Wu, C. C.; Lee, C. L.; Chen, S. L.; Lu, C. Y.; Wu, T. S.; Chang, F. R.; Wu, Y. C., Camptothecinoids from the seeds of Taiwanese Nothapodytes foetida. Molecules 2008, 13, 1361-1371.
104. Srinivas, K. V. N. S.; Das, B., 9-Methoxy-20-O-acetylcamptothecin, a minor new alkaloid from Nothapodites foetida. Biochemical Systematics and Ecology 2003, 31, (1), 85-87.
105. Ciddi, V.; Shuler, M. L., Camptothecine from callus cultures of Nothapodytes foetida. Biotechnology Letters 2000, 22, (2), 129-132.
106. Fulzele, D. P.; Satdive, R. K.; Pol, B. B., Growth and production of camptothecin by cell suspension cultures of Nothapodytes foetida. Planta Medica 2001, 67, (2), 150-152.
107. Tafur, S.; Nelson, J. D.; DeLong, D. C.; Svoboda, G. H., Antiviral components of Ophiorrhiza mungos. Isolation of camptothecin and 10-methoxycamptothecin. Lloydia 1976, 39, (4), 261-262.
108. Yamazaki, Y.; Urano, A.; Sudo, H.; Kitajima, M.; Takayama, H.; Yamazaki, M.; Aimi, N.; Saito, K., Metabolite profiling of alkaloids and strictosidine synthase activity in camptothecin producing plants. Phytochemistry 2003, 62, (3), 461-470.
109. Kitajima, M.; Fischer, U.; Nakamura, M.; Ohsawa, M.; Ueno, M.; Takayama, H.; Unger, M.; Stöckigt, J.; Aimi, N., Anthraquinones from Ophiorrhiza pumila tissue and cell cultures. Phytochemistry 1998, 48, (1), 107-111.
110. Arisawa, M.; Gunasekera, S. P.; Cordell, G. A.; Farnsworth, N. R., Plant Anticancer Agents XXI. Constituents of Merrilliodendron megacarpum*. Planta Medica 1981, 43, (12), 404-407.
111. Zhou, B. N.; Hoch, J. M.; Johnson, R. K.; Mattern, M. R.; Eng, W. K.; Ma, J.; Hecht, S. M.; Newman, D. J.; Kingston, D. G. I., Use of COMPARE Analysis to Discover New Natural Product Drugs: Isolation of Camptothecin and 9-Methoxycamptothecin from a New Source. Journal of Natural Products 2000, 63, (9), 1273-1276.
112. Dai, J. R.; Hallock, Y. F.; Cardellina, J. H.; Boyd, M. R., 20-O-β-Glucopyranosyl Camptothecin from Mostuea brunonis: A Potential Camptothecin Pro-Drug with Improved Solubility. Journal of Natural Products 1999, 62, (10), 1427-1429.
113. Kuo, Y. H.; Lo, J. M.; Chan, Y. F., Cytotoxic Components from the Leaves of Schefflera taiwaniana. Journal of the Chinese Chemical Society 2002, 49, (3), 427-431.
114. Chang, F. R.; Chao, Y. C.; Teng, C. M.; Wu, Y. C., Chemical Constituents from Cassytha filiformis II. Journal of Natural Products 1998, 61, (7), 863-866.
115. Chen, C. Y.; Wu T. Y.; Chang, F. R.; Wu, Y. C., Lignans and Kauranes from the Stems of Annona cherimola. Journal of the Chinese Chemical Society 1998, 45, (5), 629-634.
116. Tsukamoto, S.; Tomise, K.; Aburatani, M.; Onuki, H.; Hirorta, H.; Ishiharajima, E.; Ohta, T., Isolation of Cytochrome P450 Inhibitors from Strawberry Fruit, Fragaria ananassa. Journal of Natural Products 2004, 67, (11), 1839-1841.
117. Wang, J.; Shen, Y.; He, H.; Kang, W.; Hao, X., Norsesquiterpenoid and Sesquiterpenoid Glycosides from Evodia austrosinensis. Planta Medica 2005, 71, (01), 96-98.
118. Hisamoto, M.; Kikuzaki, H.; Nakatani, N., Constituents of the Leaves of Peucedanum japonicum Thunb. and Their Biological Activity. Journal of Agricultural and Food Chemistry 2004, 52, (3), 445-450.
119. Yue, J.; Lin, Z.; Wang, D.; Sun, H., A sesquiterpene and other constituents from Erigeron breviscapus. Phytochemistry 1994, 36, (3), 717-719.
120. Mizutani, K.; Yuda, M.; Tanaka, O.; Saruwatari, Y. I.; Fuwa, T.; Jia M. R.; Ling, Y. K.; Pu, X. F., Chemical Studies on Chinese Traditional Medicine, Dangshen. I. Isolation of (Z)-3- and (E)-2-Hexenyl β-D-Glucosides. Chemical & Pharmaceutical Bulletin 1988, 36, (7), 2689-2690.
121. El-Naggar, L. J.; Beal, J. L., Iridoids. A Review. Journal of Natural Products 1980, 43, (6), 649-707.
122. Recio-Iglesias, M. C.; Marston, A.; Hostettmann, K., Xanthones and secoiridoid glucosides of Halenia campanulata. Phytochemistry 1992, 31, (4), 1387-1389.
123. Shiobara, Y.; Kato, K.; Ueda, Y.; Taniue, K.; Syoha, E.; Nishimoto, N.; de Oliveira, F.; Akisue, G.; Kubota Akisue, M.; Hashimoto, G., Secoiridoid glucosides from Chelonanthus chelonoides. Phytochemistry 1994, 37, (6), 1649-1652.
124. Shen, W.; Coburn, C. A.; Bornmann, W. G.; Danishefsky, S. J., Concise total syntheses of dl-camptothecin and related anticancer drugs. The Journal of Organic Chemistry 1993, 58, (3), 611-617.
125. Richard T. Brown; Liu Jianli; Santos, C. A. M., Biogenetically patterned synthesis of camptothecin and 20-deoxycamptothecin. Tetrahedron Letters 2000, 41, 859-862.
126. Kitajima, M.; Fujii, N.; Yoshino, F.; Sudo, H.; Saito, K.; Aimi, N.; Takayama, H., Camptothecins and Two New Monoterpene Glucosides from Ophiorrhiza liukiuensis. Chemical & Pharmaceutical Bulletin 2005, 53, (10), 1355-1358.
127. Lu, Y.; Sun, Y.; Foo, L. Y.; McNabb, W. C.; Molan, A. L., Phenolic glycosides of forage legume Onobrychis viciifolia. Phytochemistry 2000, 55, (1), 67-75.
128. Ohmoto, T.; Yamaguchi, K., Constituents of Pollen. XV. Constituents of Biota orientalis (L.) ENDL. (1). Chemical & Pharmaceutical Bulletin 1988, 36, (2), 807-809.
129. Zhu, X.; Dong, X.; Wang, Y.; Ju, P.; Luo S., Phenolic Compounds from Viburnum cylindricum. Helvetica Chimica Acta 2005, 88, (2), 339-342.
130. Chiang, Y. M.; Kuo, Y. H., Two novel α-tocopheroids from the aerial roots of Ficus microcarpa. Tetrahedron Letters 2003, 44, (27), 5125-5128.
131. Roomi, M. W.; Monterrey, J. C.; Kalinovsky, T.; Rath M.; Nienzwiecki, A., Patterns of MMP-2 and MMP-9 expression in human cancer cell lines. Oncology Reports 2009, 21, (5), 1323-1333.
132. Toda, S.; Miyamoto, M.; Kinoshita, H.; Inomata, K., Selective Mono- and Bis (alkoxycarbonylation) s of Olefins Catalyzed by Palladium in the Presence of Cu(I) or Cu(II) Chloride under Remarkably Mild Conditions. Application to the Synthesis of γ-Butyrolactone Derivatives. Bulletin of the Chemical Society of Japan 1991, 64, (12), 3600-3606.
133. Durand, S.; Parrain, J. L.; Santelli, M., A Large Scale and Concise Synthesis of r-Linolenic Acid from 4-Chlorobut-2-yn-1-ol. Synthesis 1998, 1998, (07), 1015-1018.
134. Stamatov, S. D.; Stawinski, J., A Simple and Efficient Method for Direct Acylation of Acetals with Long Alkyl-Chain Carboxylic Acid Anhydrides. Tetrahedron 2000, 56, (49), 9697-9703.
135. Nikitas Ragoussis; Ragoussis, V., Improvement on the synthesis of (E)-alk-3-enoic acids. Journal of the Chemical Society, Perkin Transactions 1 1998, (21), 3529-3534.
136. Li, T. S.; Li, A. X., Montmorillonite clay catalysis. Part 10.1 K-10 and KSF-catalysed acylation of alcohols, phenols, thiols and amines: scope and limitation. Journal of the Chemical Society, Perkin Transactions 1 1998, (12), 1913-1918.
137. Ma, C. Y.; Liu, W. K.; Che, C. T., Lignanamides and Nonalkaloidal Components of Hyoscyamus niger Seeds. Journal of Natural Products 2002, 65, (2), 206-209.
138. Ahmad, V. U.; Ali, M. S., Terpenoids from marine red alga Laurencia pinnatifida. Phytochemistry 1991, 30, (12), 4172-4174.
139. Itoh, D.; Kawano, K.; Nabeta, K., Biosynthesis of Chloroplastidic and Extrachloroplastidic Terpenoids in Liverwort Cultured Cells:  13C Serine as a Probe of Terpene Biosynthesis via Mevalonate and Non-mevalonate Pathways. Journal of Natural Products 2003, 66, (3), 332-336.
140. Zhang, Y.; Jayaprakasam, B.; Seeram, N. P.; Olson, L. K.; DeWitt, D.; Nair, M. G., Insulin Secretion and Cyclooxygenase Enzyme Inhibition by Cabernet Sauvignon Grape Skin Compounds. Journal of Agricultural and Food Chemistry 2003, 52, (2), 228-233.
141. Saimaru, H.; Orihara, Y.; Tansakul, P.; Kang, Y. H.; Shibuya, M.; Ebizuka, Y., Production of Triterpene Acids by Cell Suspension Cultures of Olea europaea. Chemical & Pharmaceutical Bulletin 2007, 55, (5), 784-788.
142. Chen, K. S.; Chang, F. R.; Chia, Y. C.; Wu, T. S.; Wu, Y. C., Chemical Constituents of Neolitsea parvigemma and Neolitsea konishii. Journal of the Chinese Chemical Society 1998, 45, (1), 103-110.
143. Cheng, H. H.; Wang, H. K.; Ito, J.; Bastow, K. F.; Tachibana, Y.; Nakanishi, Y.; Xu, Z.; Luo, T. Y.; Lee, K. H., Cytotoxic Pheophorbide-Related Compounds from Clerodendrum calamitosum and C. cyrtophyllum§. Journal of Natural Products 2001, 64, (7), 915-919.
144. Magnus Cernerud; JoséAntonio Reina; Jörgen Tegenfeldt; Moberg, C., Chiral polymers via asymmetric epoxidation and asymmetric dihydroxylation. Phytochemistry 1996, 7, (10), 2863-2870.
145. Etzenhouser, B.; Hansch, C.; Kapur, S.; Selassie, C. D., Mechanism of Toxicity of Esters of Caffeic and Dihydrocaffeic Acids. Bioorganic & Medicinal Chemistry 2001, 9, (1), 199-209.
146. Wang, M.; Kikuzaki, H.; Zhu, N.; Sang, S.; Nakatani, N.; Ho, C. T., Isolation and Structural Elucidation of Two New Glycosides from Sage (Salvia officinalis L.). Journal of Agricultural and Food Chemistry 2000, 48, (2), 235-238.
147. Lee, T. H.; Chiou, J. L.; Lee, C. K.; Kuo, Y. H., Separation and Determination of Chemical Constituents in the Roots of Rhus javanica L. var. roxburghiana. Journal of the Chinese Chemical Society 2005, 52, (4), 833-841.
148. Kuo, Y. H.; Yeh, M. H., Chemical Constituents of Heartwood of Bauhinia purpurea. Journal of the Chinese Chemical Society 1997, 44, (4), 379-383.
149. Matsuda, H.; Higashino, M.; Chen, W.; Tosa, H.; Iinuma, M.; Kubo, M., Studies of Cuticle Drugs from Natural Sources. III. Inhibitory Effect of Myrica rubra on Melanin Biosynthesis. Biological & Pharmaceutical Bulletin 1995, 18, (8), 1148-1150.
150. Sawai, Y.; Moon, J. H.; Sakata, K.; Watanabe, N., Effects of Structure on Radical-Scavenging Abilities and Antioxidative Activities of Tea Polyphenols:  NMR Analytical Approach Using 1,1-Diphenyl-2-picrylhydrazyl Radicals. Journal of Agricultural and Food Chemistry 2005, 53, (9), 3598-3604.
151. Bilia, A. R.; Ciampi, L.; Mendez, J.; Morelli, I., Phytochemical investigations of Licania genus. Flavonoids from Licania pyrifolia. Pharmaceutica Acta Helvetiae 1996, 71, (3), 199-204.
152. Kazuma, K.; Noda, N.; Suzuki, M., Malonylated flavonol glycosides from the petals of Clitoria ternatea. Phytochemistry 2003, 62, (2), 229-237.
153. Yang, M. H.; Kong, L. Y., Flavonols and flavonol glycosides from Rhododendron irroratum. Chemistry of Natural Compounds 2008, 44, (1), 98-99.
154. Lee, C. K.; Chang, M. H., The Chemical Constituents from the Heartwood of Eucalyptus citriodora. Journal of the Chinese Chemical Society 2000, 47, (3), 555-560.
155. Pansare, S. V.; Jain, R. P., Enantioselective Synthesis of (S)-(+)-Pantolactone. Organic Letters 2000, 2, (2), 175-177.

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系統識別號 U0007-2207200917251600
論文名稱(中文) 台灣兩種真菌醱酵液及菌絲體之生物活性成分研究
論文名稱(英文) Studies on the bioactive constituents of the fermented broths and mycelia of two fungus species isolated in Taiwan
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 97
學期 2
出版年 98
研究生(中文) 曹雅菁
學號 M303096009
學位類別 碩士
語文別 中文
口試日期 2009-06-19
論文頁數 93頁
口試委員 委員-郭悅雄 教授
委員-吳天賞 教授
指導教授-李宗徽 教授
關鍵字(中) 真菌
誘導型一氧化氮合成酶
Psathyrella velutina
Neosartorya
psathyrone
關鍵字(英) iNOS
Psathyrella velutina
Neosartorya
psathyrone
fungus
學科別分類
中文摘要 本研究利用抑制一氧化氮合成酶 (inducible nitric oxide synthase, iNOS) 來篩選34株單離自本土的真菌,藉以找尋出具有抗發炎活性的真菌株,發現其中13株真菌的醱酵培養液或菌絲體萃取物,對於離體RAW264.7細胞產生一氧化氮具有顯著的抑制作用。後續選定其中兩株包括:Psathyrella velutina (#96110901) 及Neosartorya sp. (#325) 進行其活性成分研究,再選擇以麥芽抽取物為培養基加以擴大培養後,針對醱酵液及菌絲所含代謝物進行一系列的分析、分離、純化與結構解析,計分離出10個化合物,分別為:psathyrone (1)、5-hydroxymethylfurfural (2)、bis-2,5-hydroxymethylfuran (3) 2-(4-hydroxy-2-octyl-5-oxotetrahydrofuran-3-yl)acrylic acid (6)、2-(4-hydroxy-2-octyl-5-oxotetrahydrofuran-3-yl)acrylic acid methyl ester (7)、maltol (8)、succinic acid (9) 和fumaric acid (10),以及一混合物含 (22E,24S)-24-methylnorcholesta-5,7,22-trien-3β-ol (4) 和 (22E,24R)-24-methylnorcholesta-5,7,22-trien-3β-ol (5)(1:1),其中psathyrone (1) 是過去未曾報導過的新化合物,化合物6和7則是首次由自然界中分離得到之天然物。天然物純質進一步進行抑制iNOS試驗,結果顯示化合物6和7對於離體RAW264.7細胞產生一氧化氮具有顯著的抑制作用,其半抑制濃度 (IC50) 分別為12.17 ± 1.49和11.36 ± 0.97 μM。
英文摘要 In this study, 34 strains of fungus isolated in Taiwan were cultured for the screening of iNOS (inducible nitric oxide synthase) inhibitory activities, and among them, the fermented broths or mycelia of 13 fungus species exhibited significant activities on the inhibition of NO production in lipopolysaccharides (LPS) activated mouse macrophage RAW264.7. Based on this findings, 2 strains of fungus including Psathyrella velutina (#96110901) and Neosartorya sp. (#325) were thus selected to investigate their bioactive constituents. A series of bioassay-guided chemical examination on the fermented broth or mycelia of these two strains were carried out, which resulted in the isolation of ten compounds. Their structures were elucidated to be psathyrone (1), 5-hydroxymethylfurfural (2), bis-2,5-hydroxymethylfuran (3), 2-(4-hydroxy-2-octyl-5-oxotetrahydrofuran-3-yl)acrylic acid (6), 2-(4-hydroxy-2-octyl-5-oxotetrahydrofuran-3-yl)acrylic acid methyl ester (7), maltol (8), succinic acid (9), fumaric acid (10), and a mixture of (22E,24S)-24-methylnorcholesta-5,7,22-trien-3β-ol (4), and (22E,24R)-24-methylnorcholesta-5,7,22-trien-3β-ol (5)(1:1) on the basis of spectroscopic analysis. Of these compounds identified, psathyrone (1) was a new compound with a rare 5/6/5/6-fused ring system, and 6 and 7 were isolated for the first time from natural resources. The bioactivities of all the pure entities were further evaluated. The results of biological tests indicated that 6 and 7 exhibited significant activities on the inhibition of NO production in lipopolysaccharides (LPS) activated macrophage RAW264.7 with IC50 values of 12.17 ± 1.49 and 11.36 ± 0.97 μM, respectively.
論文目次 謝誌...................................................I
中文摘要...............................................II
Abstract...............................................IV
總目錄.................................................VI
表目錄.................................................VIII
圖目錄.................................................IX
縮寫表.................................................XII
壹、緒論與研究目的.....................................1
貳、親緣相同 (近) 真菌之天然物文獻回顧.................5
参、實驗結果與討論.....................................8
3.1.1 Psathyrone (1) 之結構解析...................13
3.1.2 5-Hydroxymethylfurfural (2) 之結構解析......23
3.1.3 Bis-2,5-hydroxymethylfuran (3) 之結構解析...27
3.1.4 (22E,24S)-24-Methylnorcholesta-5,7,22-trien-3β-ol (4) 和 (22E,24R)-24-methylnorcholesta-5,7,22-trien-3β-ol (5) 之結構解析..................................30
3.1.5 2-(4-Hydroxy-2-octyl-5-oxotetrahydrofuran-3-yl)acrylic acid (6) 之結構解析...........38
3.1.6 2-(4-Hydroxy-2-octyl-5-oxotetrahydrofuran-3-yl)acrylic acid methyl ester (7) 之結構解析....................................46
3.1.7 Maltol (8) 之結構解析.......................54
3.1.8 Succinic acid (9) 之結構解析................57
3.1.9 Fumaric acid (10) 之結構解析................60
3.2 一氧化氮 (NO) 濃度之測定結果....................63
3.3 細胞毒性測試結果................................64
3.4 討論............................................66
肆、實驗部份...........................................68
4.1 儀器設備與試劑..................................68
4.2 培養基配置......................................69
4.3 真菌菌株之萃取與分離............................69
4.3.1 #96110901培養液之成分分離與純化.............71
4.3.2 #96110901菌絲之甲醇萃取物之成分分離與純化...75
4.3.3 #325培養液之成分分離與純化..................76
4.4 一氧化氮 (NO) 濃度之測定:Griess reagent assay..78
4.5 細胞毒性測試 (Alamar Blue assay)................81
4.6 各成分之物理數據................................84
参考文獻...............................................89
參考文獻 1. Moncada, S., Palmer, R.M.J., and Higgs, E.A. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol. Rev. 1991, 43, 109-42.
2. Nathan, C. Nitric oxide as a secretory product of mammalian cells. FASEB J. 1992, 6, 3051-64.
3. MacMicking, J., Xie, Q.W., and Nathan, C. Nitric oxide and macrophage function. Annu. Rev. Immunol. 1997, 15, 323-50.
4. Wink, D.A., and Mitchell, J.B. Chemical biology of nitric oxide: Insights into regulatory, cytotoxitic, and cytoprotective mechanisms of nitric oxide. Free Rad. Biol. Med. 1998, 25, 434-56.
5. Clancy, R.M., and Abramson, S.B. Nitric oxide: a novel mediator of inflammation. Proc. Soc. Exp. Biol. Med. 1995, 210, 93-101.
6. Laskin, D.L., and Pendino, K.J. Macrophages and inflammatory mediators in tissue injury. Annu. Rev. Pharmacol. Toxicol. 1995, 35, 655-77.
7. Cochran, F.R., Selph, J., and Sherman, P. Insights into the role of nitric oxide in inflammatory arthritis. Med. Res. Rev. 1996, 16, 547-63.
8. Culotta, E., and Koshland, D.E. NO news is good news. Science 1992, 258, 1862-5.
9. Gross, S.S., and Wolin, M.S. Nitric oxide: pathophysiological mechanisms. Annu. Rev. Physiol. 1995, 57, 737-69.
10. Wolkow, P.P. Involvement and dual effects of nitric oxide in septic shock. Inflamm. Res. 1998, 47, 152-66.
11. Palmer, R.M., Ashton, D.S., and Moncada, S. Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature 1988, 333, 664-6.
12. Nathan, C. Nitric oxide as a secretory product of mammalian cells. FASEB J. 1992, 6, 3051-64.
13. Knowles, R.G., and Moncada, S. Nitric oxide synthases in mammals. Biochem J. 1994, 298, 249-58.
14. Sheng, H., Gagne, G.D., Matsumoto, T., Miller, M.F., Förstermann, U., and Murad, F. Niric oxide synthase in bovine superior cervical ganglion. J. Neurochem. 1993, 61, 1120-6.
15. Schmidt, H.H., and Walter, U. NO at work. Cell 1994, 78, 919-25.
16. Förstermann, U., and Kleinert, H. Nitric oxide synthase: expression and expressional control of the three isoforms. Naunyn Schmiedebergs Arch. Pharmacol. 1995, 352, 351-64.
17. Knowles, R.G. and Moncada, S. Nitric oxide synthases in mammals. Biochem. J. 1994, 298, 249-58.
18. Larlux, F.S., Pavlick, K.P., Hines, I.N., Kawachi, S., Harada, H., Bharwani, S., Hoffman, J.M. and Grisham, M.B. Role of nitric oxide in inflammation. Acta. Physiol. Scand. 2001, 173, 113-8.
19. Vural, C., and Gungor, A. Nitric oxide and the upper airways: recent discoveries. Journal of Ear, Nose and Throat 2003, 10, (1), 39-44.
20. Sautebin, L. Prostaglandins and nitric oxide as molecular target for anti-inflammatory therapy. Fitoterapia. 2001, 71, 48-57.
21. Sawada, K. Descriptive catalogue of Taiwan (Formosan) fungi. Part XI. Special Publ. Coll. Agric. Nat1. National Taiwan University 1959, 268.
22. 陳瑞青,臺灣菌類資源調查之歷史與現況。中央研究院植物研究所專刊第十一號,1992,119-30。
23. 王也珍、吳聲華、周文能、張東柱、陳桂玉、陳淑芬、陳城箖、曾顯雄、劉錦惠、謝文瑞、謝煥儒、鍾兆玄和簡秋源,臺灣真菌名錄。行政院農業委員會出版,1999,p. 289。
24. Morino, T., Nishimoto, M., Itou, N. and Nishikiori, T.. NK372135S, novel antifungal agents produced by Neosartoria fischeri. J. Antibiotics 1994, Dec., 1546-8.
25. Asami, Y., Kakeya, H., Onose, R., Chang, Y.H., Toi, M., and Osada, H. RK-805, an endothelial-cell-growth inhibitor produced by Neosartorya sp., and a docking model with methionine aminopeptidase-2. Tetrahedron 2004, 60, 7085-91.
26. Asami, Y., Kakeya, H., Onose, R., Yoshida, A., Matsuzaki, H., and Osada, H. A novel angiogenesis inhibitor containing a 1-oxa-7-
azaspiro[4.4]non-2-ene-4,6-dione skeleton produced by the fungus Neosartorya sp. Org. Lett. 2002, 4, (17), 2845-8.
27. Wong, S.M., Musza, L.L., Kydd, G.C., Gillum, R.K.A.M., and Cooper, R. Fiscalins: new Substance P inhibitors produced by the fungus Neosartorya fischeri. J. Antibiot. 1992, 46, (4), 545-53.
28. Ozoe, Y., Kuriyama, T., Tachibana, Y., Harimaya, K., Takahashi, N., Takashi Yaguchi, Suzuki, E., Imamura, K.I., and Oyama, K. Isocoumarin derivative as a novel GABA receptor ligand from Neosartorya quadricincta. J. Pestic. Sci. 2004, 29, (4), 328-31.
29. Proksa, B., Uhrin, D., Liptaj, T. and Sturdikova, M. Neosartorin, an ergochrome biosynthesized by Neosartorya fischeri. Phytochem. 1998, 48, (7), 1161-4.
30. Nakadate, S., Nozawa, K., Horie, H., Fujii, Y., Nagai, M., Hosoe, T., Kawai, K.I., Yaguchi, T., and Fukushima, K. Eujavanicols A-C, decalin derivatives from Eupenicillium javanicum. J. Nat. Prod. 2007, 70, 1510-2.
31. Davis, R.A., Andjic, V., Kotiw, M., and Shivas, R.G. Phomoxins B and C: polyketides from an endophytic fungus of the genus Eupenicillium. Phytochem. 2005, 66, 2771-5.
32. Ahmed, A. Acetylenes and dichloroanisoles from Psathyrella scobinacea. Phytochem. 2000, 55, 921-6.
33. Gadir, S.A., Smith, Y., Taha, A.A., and Thaller, V.. Isolation and synthesis of scobinolide, a new monoterpene lactone from cultures of the fungus Psathyrella scobinacea. J. Chem. Res. 1986, 102-3.
34. Brain, J.M., Daniel, D.S., and Andrew T. Macphail. Synthetic and kinetic studies of substituent effects in the furan intramolecular Diels-Alder reaction. Tetrahedron 1994, 50, (23), 6767-82.
35. Fawcett, A.H., Yau, T.F., Mulemwa, J.N., and Tan, C.E.. The free-radically prepared copolymers of acrylonitrile with furfuryl alcohol and similar furan derivatives. Br. Polym. J. 1987, 19, 211-21.
36. Itoh, T., Sica, D., and Djerassi, C. Isolation and structure elucidation of seventy-four sterols from the sponge Axinella cannabina. J. Chem. Soc. Perkin Trans. I 1983, 147-53.
37. Brookes, D., Tidd, B.K., and Turner, W.B. Avenaciolide, an antifungal lactone from Aspergillus avenaceus. J. Chem. Soc. 1963, 68, 5385-91.
38. Sun, P., Ye, W., Zhao, J., Pei, Y., Wang, Z., Chen, Y., Ogihara, Y., and Takeda, T. Studies on the constituents of Epimedium koreanum. Chem. Pharm. Bull. 1995, 43, (4), 703-4
39. Bal, D., Kraska-Dziadecka, A., Gradowska, W., and Gryff-Keller, A. Investigation of a wide spectrum of inherited metabolic disorders by 13C NMR spectroscopy. Acta Biochim. Pol. 2008, 55, (1), 107-18.
40. Larsen, J.W., and Bouis, P.A. Protonation of fumaric and maleic acids and their diethyl derivatives. J. Org. Chem. 1973, 38, (7), 1415-7.

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系統識別號 U0007-2307200904535000
論文名稱(中文) 臺灣三種海洋細菌之生物活性成分研究
論文名稱(英文) Studies on the bioactive constituents of three marine bacterial species isolated in Taiwan
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 97
學期 2
出版年 98
研究生(中文) 許瑜真
學號 M303096005
學位類別 碩士
語文別 中文
口試日期 2009-06-19
論文頁數 208頁
口試委員 指導教授-李宗徽 博士
委員-郭悅雄 博士
委員-吳天賞 博士
關鍵字(中) 海洋細菌
關鍵字(英) Pseudomonas aeruginosa
Actinomycin X2
diketopiperazine
NOS
學科別分類
中文摘要 以三種不同培養基篩選出143株海洋細菌株,經醱酵培養後,萃取液初步以抑制一氧化氮合成酶 (NOS) 的活性與利用Phenylephrine誘發大鼠 (Sprague-Dawley Rat) 胸主動脈收縮活性平台進行篩選出具有抑制血管收縮的菌株。結果自143株海洋菌株萃取液中發現二株對於RAW 264.7細胞具有細胞毒性,分別為 Streptomyces sp. (#HYC21) 和 Pseudoalteromonas sp. (#HYC13),另一株 Pseudomonas aeruginosa (#M1B) 則具有抑制血管收縮的活性,後續針對這三株具有生物活性的菌株以PPY、PY和PYG 培養基分批擴大培養,進行一系列的分析、分離、純化與構造解析,計分離、決定出二十一個化合物,包括:二個環肽類抗生素 (cyclic peptide antibiotics) 的actinomycin X2 (1)、actinomycin D (2);二個多溴酚類抗生素 (polybrominated phenolic antibiotics) 的 pentabromopseudilin (8) 和3,3′,5,5′-tetrabromo-2,2′-biphenyldiol (9);三個膽酸及其類似物 (cholic acid and its analogs) 的cholic acid (10)、deoxycholic acid (11)、glycocholic acid (12);五個喹啉生物鹼 (quinoline alkaloid) 分別為2- heptylquinol-4-one (13)、2-heptyl-3-hydroxyquinolin-4(1H)-one (14)、2-(1′E-nonenyl)quinol-4-one (15)、2-nonylquinol-4-one (16)、 3-heptyl-3-hydroxy-1,2,3,4-tetrahydroquinoline-2,4-dione (17);二個酚類化合物 (phenolics) 的2-(2-hydroxyphenyl)-2-thiazoline-4-methanol (18)、4,4′-isopropylidenebisphenol (19);一個吩嗪生物鹼 (phenazine alkaloid) phenazine-1-carboxamide (21) 及五個環化雙胺基酸 (diketopiperazine) 分別為cyclo-L-Pro-L-Val (3)、cyclo-L-Pro-L-Ile (4)、cyclo-L-Pro-L-Leu (5)、cyclo-D-Pro-L-Phe (6)、cyclo-L-Pro-L-Phe (7)、cyclo-L-Pro-L-Trp (20)。此外,由M1B所分離得到的化合物在生物活性及作用機制,刻正持續探究中。
英文摘要 In this study, 143 strains of marine bacteria isolated from Taiwan were cultured for the screening of their inducible nitric oxide synthase (iNOS) inhibitory activity and vasorelaxing activity. Of these bacterial strains monitored, HYC21 (Streptomyces sp.) and HYC13 (Pseudoalteromonas sp.) exhibited cytotoxic activities against RAW 264.7 cell line. While M1B (Pseudomonas aeruginosa) exhibited vasorelaxing activity on Sprague-Dawley rats induced by phenylephrine. Based on these findings, the three strains were thus mass cultured and a series of separation and isolation were undertaken to investigate their active principles. Totally twenty-one compounds including 2 cyclic peptide antibiotics, 2 polybrominated phenolic antibiotics, 3 cholic acid and its analogs, 5 quinoline alkaloids, 2 phenolics, a phenazine alkaloid, and 6 diketopiperazine was isolated and identified. Their structures were elucidated to be actinomycin X2 (1), actinomycin D (2), cyclo-L-Pro-L-Val (3), cyclo-L-Pro-L-Ile (4), cyclo-L-Pro-L-Leu (5), cyclo-D-Pro-L-Phe (6), cyclo-L-Pro-L-Phe (7), pentabromopseudilin (8), 3,3′,5,5′-tetrabromo-2,2′-biphenyldiol (9), cholic acid (10), deoxycholic acid (11), glycocholic acid (12), 2-heptylquinol-4-one (13), 2-heptyl-3-hydroxyquinolin-4(1H)-one (14), 2- (1′E-nonenyl)quinol-4-one (15), 2-nonylquinol-4-one (16), 3- heptyl-3-hydroxy-1,2,3,4-tetrahydroquinoline-2,4-dione (17), 2-(2-hydroxyphenyl)-2-thiazoline-4-methanol (18), 4,4′-isopropylidenebisphenol (19), cyclo-L-Pro-L-Trp (20), phenazine-1-carboxamide (21).
論文目次 第一章 緒論與研究目的1
1.1海洋天然物研究的發展近況 1
1.2三種海洋細菌天然物之文獻回顧 8
第二章 實驗結果與討論 29
2.1化合物成分之結構解析 40
2.1.1 Actinomycin X2 (1) 40
2.1.2 Actinomycin D (2) 52
2.1.3 Pentabromopseudilin (8) 60
2.1.4 3,3?S,5,5?S-Tetrabromo-2,2?S-biphenyldiol (9) 64
2.1.5 Cholic acid (10) 68
2.1.6 Deoxycholic acid (11) 79
2.1.7 Glycocholic acid (12) 89
2.1.8 2-Heptylquinolin-4(1H)-one (13) 99
2.1.9 2-Heptyl-3-hydroxyquinolin-4(1H)-one (14) 103
2.1.10 2-(1′E-Nonenyl)-quinol-4-one (15) 109
2.1.11 2-Nonylquinolin-4(1H)-one (16) 114
2.1.12 3-Heptyl-3-hydroxyquinoline-2,4(1H,3H)-dione
(17) 118
2.1.13 4-Hydroxymethyl-2-(o-hydroxyphenyl)thiazoline
(18) 122
2.1.14 4,4?S-Isopropylidenediphenol (19) 128
2.1.15 Cyclo-L-Pro-L-Trp (20) 132
2.2海洋菌株DNA鑑定結果 142
2.2.1 #HYC21 (Streptomyces sp.) 142
2.2.2 #HYC13 (Pseudoalteromonas sp.) 146
2.2.3 #M1B (Pseudomonas aeruginosa) 148
2.3海洋菌株活性檢測結果 150
2.3.1 143株海洋細菌粗萃物NOS活性初篩檢測結果 150
2.3.2 #M1B抑制抗藥性金黃色葡萄球菌生長之活性檢測
結果 153
2.4討論 154
第三章 實驗部份 156
3.1儀器設備與試劑 156
3.2樣品緩衝液配置 158
3.3培養基配置 158
3.4 16S rDNA基因分析 161
3.5抑菌特性測定 165
3.5.1細菌的計數 165
3.5.2瓊脂紙錠擴散試驗 (disc agar-diffusion test) 166
3.6一氧化氮合成酶活性實驗167
3.6.1 RAW 264.7 細胞產生一氧化氮的測定實驗 167
3.6.2 RAW 264.7 細胞存活率測定實驗 168
3.7離體SD大白鼠胸主動脈血管環張力實驗 169
3.7.1胸主動脈血管環的製備 169
3.7.2 SD大白鼠血管環張力實驗 169
3.8海洋菌株之篩選、萃取與分離 171
3.8.1#HYC21之成分分離、純化 173
3.8.2#HYC13之成分分離、純化 176
3.8.3#M1B之成分分離、純化 180
3.9各成分之物理數據 186
參考文獻 198


參考文獻 1. Castro, P.,Huber, M. E., In Marine biology, McGraw-Hill, Dubuque, IA: 2005; Vol. 5th edn.
2. Roberts, J. M., Wheeler, A. J.,Freiwald, A., Reefs of the deep: the biology and geology of cold-water coral ecosystems. Science 2006, 312, 543-547.
3. Brandt, A., Gooday, A. J., Brandão, S. N., Brix, S., Brökeland, W., Cedhagen, T., Choudhury, M., Cornelius, N., Danis, B., Mesel, I. D., Diaz, R. J., Gillan, D. C., Ebbe, B., Howe, J. A., Janussen, D., Kaiser, S., Linse, K., Malyutina, M., Pawlowski, J., Raupach, M.,Vanreusel, A., First insights into the biodiversity and biogeography of the Southern Ocean deep sea. Nature 2007, 447, 307-311.
4. Sibuet, M.,Olu, K., Biogeography, biodiversity and fluid dependence of deep-sea cold-seep communities at active and passive margins. Deep Sea Res. Part II: Topical Studies Oceanograph 1998, 45, 517-567.
5. Venter, J. C., Remington, K., Heidelberg, J. F., Halpern, A. L., Rusch, D., Eisen, J. A., Wu, D. Y., Paulsen, I., Nelson, K. E., Nelson, W., Fouts, D. E., Levy, S., Knap, A. H., Lomas, M. W., Nealson, K., White, O., Peterson, J., Hoffman, J., Parsons, R., Baden-Tillson, H., Pfannkoch, C., Rogers, Y. H.,Smith, H. O., Environmental genome shotgun sequencing of the Sargasso Sea. Science 2004, 304, 66-74.
6. Grassle, J. F.,Maciolek, N. J., Deep-sea species richness: regional and local diversity estimates from quantitative bottom samples. American Naturalist 1992, 139, 313-341.
7. Gage, J. D., Why are there so many species in deep-sea sediments? Journal of Experimental Marine Biology and Ecology 1996, 200, 257-286.
8. Snelgrove, P. V. R.,Smith, C. R., A riot of species in an environmental calm: the paradox of the species-rich deep-sea floor. Oceanography and Marine Biology 2002, 40, 311-342.
9. Newman, D. J., Cragg, G. M.,Snader, K. M., Natural Products as Sources of New Drugs over the Period 1981-2002. Journal of Natural Products 2003, 66, 1022-1037.
10. Zobell, C. E., In Marine microbiology, a monograph on hydrobacteriology, Waltham, Mass., Chronica botanica company: 1946.
11. Goodfellow, M.,Haynes, J. A., In Actinomycetes in marine sediments. Biological, Biochemical, and Biomedical Aspects of Actinomycetes, Ortiz-Ortiz, L.,L. F. Bojalil, V. Yakoleff, Eds. 1984; pp 453-472.
12. Lovell, F. M., The Structure of a Bromine-Rich Marine Antibiotic. Journal of the America Chemical Society 1966, 88, 4510-4511.
13. Burkholder, P. R., Pfister, R. M.,Leitz, F. H., Production of a pyrrole antibiotic by a marine bacterium. Applied and Environmental Microbiology 1966, 14, 649-653.
14. Moore, B. S., Biosynthesis of marine natural products: microorganisms and macroalgae. Nat. Prod. Rep. 1999, 16, 653-74.
15. Ruggieri, G. D., Drugs from the sea. Science 1976, 194, (4264), 491-497.
16. Lebar, M. D., Heimbegner, J. L.,Baker, B. J., Cold-water marine natural products. Natural Product Reports 2007, 24, 774.
17. Nicholasa, G. M.,Phillips, A. J., Marine natural products: synthetic aspects. Natural Product Reports 2006, 23, 79.
18. Pietra, F., Secondary metabolites from marine microorganisms: bacteria, protozoa, algae and fungi. Achievements and prospects. Natural Product Reports 1997, 14, 453.
19. Simmons, T. L., Andrianasolo, E., Mcphail, K., Flatt, P.,Gerwick, W. H., Marine natural products as anticancer drugs. Molecular Cancer Therapeutics 2005, 4, (2), 333-342.
20. Bernan, V. S., Greenstein, M.,Maise, W. M., Marine microorganisms as a source of new natural products. Advances in Applied Microbiology 1997, 43, 57.
21. Blunt, J. W., Copp, B. R., Hu, W.-P., Munro, M. H. G., Northcote, P. T.,Prinsep, M. E. R., Marine natural products. Natural Product Reports 2009, 26, 170-244.
22. Faulkner, D. J., Marine natural products. Natural Product Reports 1999, 16, 155-198.
23. Solanki, R., Khanna, M.,Lal, R., Bioactive compounds from marine actinomycetes. Indian Journal of Microbiology 2008, 48, 410–431.
24. Maldonado, L. A., Stach, J. E. M., Pathom-Aree, W., Ward, A. C., Bull, A. T.,Goodfellow, M., Diversity of cultivable actinobacteria in geographically widespread marine Sediments. Antonie van Leeuwenhoek 2005, 87, 11-18.
25. Lam, K. S., Discovery of novel metabolites from marine actinomycetes. Current Opinion in Microbiology 2006, 9, 245-251.
26. Bowman, J. P., Bioactive Compound Synthetic Capacity and Ecological Significance of Marine Bacterial Genus Pseudoalteromonas. Marine Drugs 2007, 5, 220-241.
27. Collier, D. N., Anderson, L., Mcknight, S., Noah, T. L., Knowles, M., Boucher, R., Schwab, U., Gilligan, P.,Pesci, E. C., A bacterial cell to cell signal in the lungs of cystic fibrosis patients. FEMS Microbiology Letters 2002, 215, 41-46.
28. Baumann, U., Stocklossa, C., Greiner, W., Schulenburg, J. G. V. D.,Hardt, H. V. D., Cost of care and clinical condition in paediatric cystic fibrosis patients. Journal of Cystic Fibrosis 2003, 2, (2), 84-90.
29. Stead, P., Rudd, B. A., Bradshaw, H., Noble, D.,Dawson, M. J., Induction of phenazine biosynthesis in cultures of Pseudomonas aeruginosa by L-N-(3-oxohexanoyl) homoserine lactone. FEMS Microbiology Letters 1996, 140, (1), 15-22.
30. Gerber, N. N., Microbial Products, CRC Press, eds. In Handbook of Microbiology. , 1973; Vol. 3, pp 329-332.
31. Kock, I., Maskey, R. P., Biabani, M. A. F., Helmke, E.,Laatsch, H., 1-hydroxy-1-norresistomycin and resistoflavine methyl ether new antibiotics from marine derived Streptomycetes. Journal of Antibiotics 2005, 58, 530-534.
32. Gorajana, A., Venkatesan, M., Vinjamuri, S., Kurada, B. V., Peela, S., Jangam, P., Poluri, E.,Zeeck, A., Resistoflavine cytotoxic compound from a marine actinomycete, Streptomyces chibaensis AUBN(1)/7. Microbiological Research 2006, 29.
33. Adinaryan, G., Venkateshan, M. R., Bpiraju, V. V., Sujatha, P., Premkumar, J., Ellaiah, P.,Zeeck, A., Cytotoxic compounds from the marine actinobacterium. Bioorg Khim 2006, 32, 328-334.
34. Shiono, Y., Shiono, N., Seo, S., Oka, S.,Yamazaki, Y., Effects of polyphenolic anthrone derivatives resistomycin and hypericin on apoptois in human megakaryoblastic leukemia CMK-7cell2. Natuforsch 2002, 57, 923-929.
35. Maskey, R. P., Helmke, E.,Laatsch, H., Himalomycin A and B isolation and structure elucidation of new fridamycin type antibiotics from a marine Streptomyces isolate. Journal of Antibiotics 2003, 56, 942-949.
36. Itoh, T., Kinoshita, M., Aoki, S.,Kobayashi, M., Komodoquinone A, a novel neuritogenic anthracycline from marine Streptomyces sp. KS3. Journal of Natural Products 2003, 66, 1373-1377.
37. Macherla, V. R., Liu, J., Bellows, C., Teisan, S., Nicholson, B., Lam, K. S.,Potts, B. C. M., Glaciapyrroles A, B and C pyrrolosesquiterpenes from a Streptomyces sp. isolated from an Alaskan marine sediment. Journal of Natural Products 2005, 68, 780-783.
38. Cho, J. Y., Kwon, H. C., Williams, P. G., Kauffman, C. A., Jensen, P. R.,Fenical, W., Actinofuranones A and B, polyketides from a marine derived bacterium related to the genus Streptomyces (Actinomycetales). Journal of Natural Products 2006, 69, 425-428.
39. Asolkar, R. N., Jensen, P. R., Kauffman, C. A.,Fenical, W., Daryamides A-C weakly cytotoxic polyketides from a marine derived actinomycete of the genus Streptomyces strain CNQ-085. Journal of Natural Products 2006 69, (1), 1756-1759.
40. Moore, B. S., Trischman, J. A., Seng, D., Kho, D., Jensen, P. R.,Fenical, W., Salinamides, anti-inflammatory depsipeptides from a marine Streptomycete. J Org Chem 1999, 64, 1145-1150.
41. Miller, E. D., Kauffman, C. A., Jensen, P. R.,Fenical, W., Piperazimycins cytotoxic hexadepsipeptides from a marine derived bacterium of the genus Streptomyces. Journal of Organic Chemistry 2007, 72, 323-330.
42. Renner, M. K., Shen, Y. C., Cheng, X. C., Jensen, P. R., Frankmoelle, W., Kauffman, C. A., Fenical, W., Lobkovsky, E.,Cladry, J., Cyclomarins A-C, new anti infl ammatory cyclic peptides produced by a marine bacterium (Streptomyces sp.). Journal of the American Chemical Society 1999, 121, 11273-1276.
43. Lee, H. S., Shin, H. J., Jang, K. H., Kim, T. S., Oh, K. B.,Shin, J., Cyclic peptides of the Nocardamine class from a marine derived bacterium of the genus Streptomyces. Journal of Natural Products 2005, 68, 623-625.
44. Cho, J. Y., Kwon, H. C., Williams, P. G., Jensen, P. R.,Fenical, W., Azamerone, a terpenoid phthalazinone from a marine derived bacterium related to the genus Streptomyces (Actinomycetales) Organic Letters 2006, 8, 2471–2474.
45. Li, F., Maskey, R. P., Qin, S., Sattler, I., Fiebig, H. H., Maier, A., Zeeck, A.,Laatsch, H., Chinikomycins A and B Isolation, structure elucidation and biological activity of novel antibiotics from a marine Streptomyces sp. isolate MO45. Journal of Natural Products 2005, 68, 349–353.
46. Ward, S. L., Hu, Z., Schirmer, A., Reid, R., Revill, W. P., Reeves, C. D., Petrakovsky, O. V., Dong, S. D.,Katz, L., Chalcomycin Biosynthesis Gene Cluster from Streptomyces bikiniensis: Novel Features of an Unusual Ketolide Produced through Expression of the chm Polyketide Synthase in Streptomyces fradiae. Antimicrobial Agents and Chemotherapy 2004, 4703–4712.
47. Mitchell, S. S., Nicholson, B., Teisan, S., Lam, K. S.,Potts, B. C., Aureoverticillactam, a novel 22-atom macrocyclic lactam from the marine actinomycete Streptomyces aureoverticillatus. Journal of Natural Products 2004, 67, 1400–1402.
48. Lo′Pez, J. M. S. N., Insua, M. M. N., Baz, J. P. R., Puentes, J. L. F. N.,Herna′Ndez, L. M. C. E., New Cytotoxic Indolic Metabolites from a Marine Streptomyces Journal of Natural Products 2003, 66, 863-864.
49. Imada, C., Enzyme Inhibitors of Marine Microbial Origin with Pharmaceutical Importance. Marine Biotechnology 2004, 6, 193–198.
50. Aoyama, T., Kojima, F., Imada, C., Muraoka, Y., Naqanawa, H., Okami, Y., Takeuchi, T.,Aoyaqi, T., Pyrostatins A and B, new inhibitors of N-acetyl-beta-D-glucosamidase, produced by Streptomyces sp. SA3501 Journal of Enzyme Inhibition 1995, 8, 223–232.
51. Aoyagi, T., M.Hatsu, Imada, C., Naganawa, H., Okami, Y.,Takeuchi, T., Pyrizinostatin: a new inhibitor of pyroglutamyl peptidase Journal of Antibiotics 1992, 45, 1795–1796.
52. Manam, R. R., Teisan, S., White, D. J., Nicholson, B., Grodberg, J., Neuteboom, S. T. C., Lam, K. S., Mosca, D. A., Lloyd, G. K.,Potts, B. C. M., Lajollamycin, a nitro-tetraene spiro-b-lactone-g-lactam antibiotic from the marine actinomycete Streptomyces nodosus. Journal of Natural Products 2005, 68, 240-243.
53. Maskey, R. P., Helmke, E., Kayser, O., Fiebig, H. H., Maier, A., Busche, A.,Laatsch, H., Anticancer and antibacterial trioxacarcins with high anti-malaria activity from a marine Streptomycete and their absolute stereochemistry. Journal of Antibiotics 2004, 57, 771–779.
54. Maskey, R. P., Sevvana, M., Uson, I., Helmke, E.,Laatsch, H., Gutingimycin: a highly complex metabolite from a marine streptomycete. Angewandte Chemie International Edition 2004, 43, 1281-1283.
55. Stritzke, K., Schulz, S., Laatsch, H., Helmke, E.,Beil, W., Novel caprolactones from a marine Streptomycete. Journal of Natural Products 2004, 67, 395–401.
56. Wu, S. J., Fotso, S., Li, F., Qin, S.,Laatsch, H., Amorphane sesquiterpenes from a marine Streptomyces sp. Journal of Natural Products 2007, 70, 304–306.
57. Gerber, N. N.,Gauthier, M. J., New prodigiosin-like pigment from Alteromonas rubra. Applied and Environmental Microbiology 1979, 37, 1176-1179.
58. Kawauchi, K., Shibutani, K., Yagisawa, H., Kamata, H., Nakatsuji, S., Anzai, H., Yokoyama, Y., Ikegami, Y., Moriyama, Y.,Hirata, H., A possible immunosuppressant, cycloprodigiosin hydrochloride, obtained from Pseudoalteromonas denitrificans Biochemical and Biophysical Research Communications 1997, 237, 543-547.
59. Mitova, M., Tutino, M. L., Infusini, G., Marino, G.,Rosa, S. D., Extracellular peptides from Antarctic psychrophile Pseudoalteromonas haloplanktis. Marine Biotechnology 2005, 7, 523-531.
60. Kalinovskaya, N. I., Ivanova, E. P., Alexeeva, Y. V., Gorshkova, N. M., Kuznetsova, T. A., Dmitrenok, A. S.,Nicolau, D. V., Low-molecular-weight, biologically active compounds from marine Pseudoalteromonas species. Current Microbiology 2004, 48, (6), 441-446.
61. Andersen, R. J., Wolfe, M. S.,Faulkner, D. J., Autotoxic antibiotic production by a marine Chromobacterium. Marine Biology 1974, 27, 281-285.
62. Faulkner, D. J.,Sammes, E. G., Topics in Antibiotic Chemistry. John Wiley: New York 1978, 2, 9-58.
63. Gauthier, M. J., Validation of the name Alteromonas luteoviolacea. International Journal of Systematic Bacteriology 1982, 32, 82-86.
64. Gauthier, M. J.,Flatau, G. N., Antibacterial activity of marine violet-pigmented Alteromonas with special reference to the production of brominated compounds. Canadian Journal of Microbiology 1976, 22, 1612-1619.
65. Gribble, G. W., The diversity of naturally occurring organobromine compounds. Chemical Society Reviews 1999 28, 335-348.
66. Sakata, T., Sakaguchi, K.,Kakimoto, D., Antiobiotic production by marine pigmented bacteria I Antibacterial effect of Alteromonas luteoviolaceus. Mem. Fac Fish., Kagoshima Univ. 1982, 31, 243-250.
67. Sakata, T., Sakaguchi, K.,Kakimoto, D., Antiobiotic production by marine pigmented bacteria. II. Purification and characterization of antibiotic substances of Alteromonas luteoviolacea. . Mem Fac Fish, Kagoshima Univ 1986, (35), 29-37.
68. Jiang, Z., Boyd, K. G., Andrew, M. S., Adams, D. R., Wright, P. C.,Burgess, J. G., Two diketopiperazines and halogenated phenol from cultures of the marine bacterium, Pseudoalteromonas luteoviolacea. Natural Product Letters 2000, 14, 435-440.
69. Sobolevskaya, M. P., Smetanina, O. F., Speitling, M., Shevchenko, L. S., Dmitrenok, P. S., Laatsch, H., Kuznetsova, T. A., Ivanova, E. P.,Elyakov, G. B., Controlling production of brominated cyclic depsipeptides by Pseudoalteromonas maricaloris KMM 636T. Letters in Applied Microbiology 2005, 40, 243–248.
70. Isnansetyo, A.,Kamei, Y., MC21-A, a bactericidal antibiotic produced by a new marine bacterium, Pseudoalteromonas phenolica sp. nov. O-BC30T against methicillin resistant Staphylococcus aureus. Antimicrobial Agents and Chemotherapy 2003, 47, (2), 480-488.
71. Gallacher, S.,Birkbeck, T. H., A tissue-culture assay for direct detection of sodium-channel blocking toxins in bacterial culture supernates. FEMS microbiology letters 1992, 92, 101-108.
72. Simidu, U., Kita-Tsukamoto, K., Yasumoto, T.,M, Y., Taxonomy of marine bacteria that produce tetrodotoxin. International Journal of Systematic Bacteriology 1990, 40, 331-336.
73. Franks, A., Haywood, P., Holmström, C., Egan, S., Kjelleberg, S.,Kumar, N., Isolation and structure elucidation of a novel yellow pigment from the marine bacterium Pseudoalteromonas tunicata. Molecules 2005, 10, 1286-1291.
74. Veselova, M. A., Klein, S., Bass, I. A., Lipasova, V. A., Metlitskaya, A. Z., Ovadis, M. I., Chernin, L. S.,Khmel, I. A., Quorum sensing systems of regulation, synthesis of phenazine antibiotics, and antifungal activity in rhizospheric bacterium pseudomonas chlororaphis 449. Russian Journal of Genetics 2008, 44, 1400–1408.
75. Ligon, J. M., Hill, D. S., Hammer, P. E., Torkewitz, N. R., Hofmann, D., Kempf, H. J.,Pe′E, K. H. V., Natural products with antifungal activity from Pseudomonas biocontrol bacteria. Pest Management Science 2000, 56, 688-695.
76. Holden, M. T. G., Chhabra, S. R., Nys, R. D., Stead, P., Bainton, N. J., Hill, P. J., Manefield, M., Kumar, N., Labatte, M., England, D., Rice, S., Givskov, M., Salmond, P. C., Stewart, G. S. A. B., Bycroft, B. W., Kjelleberg, S.,Williams, P., Quorumsensing cross talk: isolation and chemical characterization of cyclic dipeptides from Pseudomonas aeruginosa and other gram-negative bacteria. Molecular Microbiology 1999 33, (6), 1254-1266.
77. Leisinger, T.,Margraff, R., Secondary metabolites of the fluorescent pseudomonads. Microbiological reviews 1979, 43, (3), 422-442.
78. Debitus, C., Guella, G., Mancini, I. I., Waikedre, J., Guemas, J. P., Nicolas, J. L.,Pietra, F., Quinolones from a bacterium and tyrosine metabolites from its host sponge, Suberea creba from the Coral Sea. Journal of Marine Biotechnology 1998, 6, (3), 136-141.
79. Yang, W., Dostal, L.,Rosazza, J. P. N., Aeruginol [ 2 -( 2 '- hydroxyphenyl )- 4 -hydroxymethylthiazole ], a new secondary metabolite from Pseudomonas aeruginosa. Journal of Natural Products 1993, 56, (11), 1993-1994.
80. Gerber, N. N., In Handbook of Microbiology, 1973; Vol. 3, pp 329-332.
81. Angell, S., Bench, B. J., Williams, H.,Watanabe, C. M. H., Pyocyanin Isolated from a Marine Microbial Population: Synergistic Production between Two Distinct Bacterial Species and Mode of Action. Chemistry & Biology 2006, 13, 1349–1359.
82. De Souza, J. T.,Raaijmakers, J. M., Polymorphisms within the prnD and pltC genes from pyrrolnitrin and pyoluteorin-producing Pseudomonas and Burkholderia spp. FEMS Microbiology Ecology 2003, 43, (1), 21-34.
83. Reimmann, C., Patel, H. M., Serino, L., Barone, M., Walsh, C. T.,Haas, D., Essential PchG-dependent reduction in pyochelin biosynthesis of Pseudomonas aeruginosa. Journal of Bacteriology 2001, 183, (3), 813-820.
84. Marchand, P. A., Weller, D. M.,Bonsall, R. F., Convenient Synthesis of 2,4-Diacetylphloroglucinol, a Natural Antibiotic Involved in the Control of Take-All Disease of Wheat Journal of Agricultural and Food Chemistry 2000, 48 (5), 1882-1887.
85. Pohanka, A., Levenfors, J.,Broberg, A., Antimicrobial Dialkylresorcinols from Pseudomonas sp. Ki19. Journal of Natural Products 2006, 69 (4), 654-665.
86. Bitzer, J., Gesheva, V.,Zeeck, A., Actinomycins with Altered Threonine Units in the β-Peptidolactone. Journal of Natural Products 2006, 69, (8), 1153-1157.
87. Arison, B. H.,Hoogsteen, K., Nuclear magnetic resonance spectral studies on actinomycin D. Preliminary observations on the effect of complex formation with 5'-eoxyguanylic acid. Biochemistry 1970, 9, (20), 3976-83.
88. Booth, H., Mauger, A. B.,Rzeszotarski, W. J., A 13C NMR Study of Actinomycin D and Related Model Peptides. Organic Magnetic Resonance 1976, 8, 219-223.
89. Cho, S. E., Goo, Y. M.,Kim, K. J., Identification of Actinomycins by High Performance Liquid Chromatography and Fast Atom Bombardment Mass Spectrometry. Archives of Pharmacal Research 1994, 17, (6), 424-427.
90. Brockmann, H., Bohnsack, G.,Gröne, H., Der Aminosäuregehalt der Actinomycine Die Naturwissenschaften 1953, 40, 223-224.
91. Johnson, A. W.,Mauger, A. B., The Isolation and Structure of Actinomycins II and III. Biochemical Journal 1959, 73, 539-553.
92. Mirau, P. A.,Shafer, R. H., High-Resolution Proton Nuclear Magnetic Resonance Analysis of Conformational Properties of Biosynthetic Actinomycin Analogues. Biochemistry 1982, 21, (11), 2622-2626.
93. Iida, T.,Chang, F. C., Potential bile acid metabolites. 6. Stereoisomeric 3,7-dihydroxy-5.beta.-cholanic acids. Journal of Organic Chemistry 1982, 47, (15), 2966-2972.
94. Waterhous, D. V., Barnes, S.,Muccio, D. D., Nuclear magnetic resonance spectroscopy of bile acids. Development of two-dimensional NMR methods for the elucidation of proton resonance assignments for five common hydroxylated bile acids, and their parent bile acid, SP-cholanoic acid. Journal of Lipid Research 1985, 26, (9), 1068-1078.
95. Ijare, O. B., Somashekar, B. S., Jadegoud, Y.,Gowda, G. A. N., 1H and 13C NMR Characterization and Stereochemical Assignments of Bile Acids in Aqueous Media. Lipids 2005, 40, (10), 1031-1041.
96. Mahout, M., Mereiter, K.,Hammerschmidt, F., Rearangement of 4-(Alkoxycarbonyloxy)quinoline N-Oxides Anna W to 4(1H)-Quinolonesoschek, Synthesis of 2-Heptyl-1-hydroxy-4(1H)-quinolone-Unexpected Rearrangement of 4-(Alkoxycarbonyloxy)quinoline N-Oxides to 1-(Alkoxycarbonyloxy)-4(1H)-quinolones. Synthesis 2007, 10, 1517–1522.
97. Pesci, E. C., Milband, J. B. J., Pearson, J. P., Mcknight, S., Kende, A. S., Greenberg, E. P.,Iglewski, B. H., Quinolone signaling in the cell-to-cell communication system of Pseudomonas aeruginosa. Proc. Natl. Acad. Sci. USA 1999, 96, 11229-11234.
98. Zunnundzhanov, A., Bessonova, I. A., Abdullaev, N. D.,Ogai, D. K., Structure of aerugine from Pseudomonas aeruginosa. Chemistry of Natural Compounds 1987, 23, 461-465.
99. Lee, J. Y., Moon, S. S.,Hwang, B. K., Isolation and Antifungal and Antioomycete Activities of Aerugine Produced by Pseudomonas fluorescens Strain MM-B16. Applied and Environmental Microbiology 2003, 69, (4), 2023-2031.
100. Subramaniam, G., Savithri, R.,Thambipillai, S., Synthesis and Antifungal Activity of Bisphenolic Derivatives. Journal of the Indian Chemical Society 1989 66, 797-799.
101. Grundmann, A.,Li, S. M., Overproduction, purification and characterization of FtmPT1, a brevianamide F prenyltransferase from Aspergillus fumigatus. Microbiology 2005, 151, 2199–2207.
102. Lin, Z.-J., Lu, X.-M., Zhu, T.-J., Fang, Y.-C., Gu, Q.-Q.,Zhu, W., GPR12 Selections of the Metabolites from an Endophytic Streptomyces sp. Asociated with Cistanches deserticola. Archives of Pharmacal Research 2008, 31, (9).
103. Grant, G. D., Hunt, A. L., Milne, P. J., Roos, H. M.,Joubert, J. A., The structure and conformation of the tryptophanyl diketopiperazines cyclo(Trp–Trp).C2H6SO and cyclo(Trp–Pro) . Journal of Chemical Crystallography 1999, 29, (4).
104. Waksman, S. A., The Actinomycetes. The Williams and Wilkins Co., Baltimore 1961, 2.
105. Minxwe, T. J., Jwnawn, P. R., Kauffman, C. A.,Fenical, W., Widespread and Persistent Populations of a Major New Marine Actinomycete Taxon in Ocean Sediments. Applied and Environmental Microbiology 2002, 68, (10), 5005-5011
106. De Beer, E. J.,Sherwood, M. B., The Paper-Disc Agar-Plate Method for the Assay of Antibiotic Substances. Journal of Bacteriology 1945, 50, (4), 459-467.
107. Adamczeski, M., Reed, A. R.,Crews, P., New and known diketopiperazines from the Caribbean sponge, Calyx cf. podatypa. Journal of Natural Products 1995, 58, 201-208.
108. Jayatilake, G. S., Thornton, M. P., Leonard, A. C., Grimwade, J. E.,Baker, B. J., Metabolites from an Antarctic sponge-associated bacterium, Pseudomonas aeruginosa. Journal of Natural Products 1996, 59, 293-296.




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系統識別號 U0007-2307200916410500
論文名稱(中文) 六種葡萄科植物對抗氧化及抗乳癌細胞株毒殺試驗效用之探討
論文名稱(英文) Antioxidant and anti-breast cancer cell activity of six Vitaceae plants
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 97
學期 2
出版年 98
研究生(中文) 涂聿杏
學號 M303096014
學位類別 碩士
語文別 中文
口試日期 2009-07-14
論文頁數 60頁
口試委員 指導教授-梁文俐
委員-楊玲玲
委員-呂鋒洲
委員-張睌E
委員-吳姿樺
關鍵字(中) 葡萄科
抗氧化
乳癌
MCF-7
關鍵字(英) Viteaceae
anti-oxidant
breast cancer
MCF-7
學科別分類
中文摘要 葡萄科植物是具清熱效果的藥材,一般民間用法為消炎、治療肝病或搗敷治癰腫。本論文以市面常見的六種葡萄科植物做抗氧化及人類乳癌細胞株MCF-7毒殺試驗為主。
在利用不同溶媒萃取葡萄科植物評估抗乳癌細胞株MCF-7毒殺試驗和抗氧化試驗中,其包含清除DPPH自由基、螯合亞鐵離子、抑制超氧陰離子之生成、還原三價鐵離子能力測定及抗肝脂質過氧化等幾項。在清除DPPH自由基方面結果顯示,用乙醇及50%乙醇萃取的葡萄科植物具有不錯效用,其中50%乙醇萃取細本山葡萄(枝)在還原能力測定及螯合亞鐵離子實驗中皆達到50%以上的抑制率。至於抑制超氧陰離子之生成方面47個樣品則都無明顯效用。本研究整體來看,利用50%乙醇萃取的葡萄科植物在抗氧化方面都比用其他溶媒更有效用。在抗氧化實驗中:細本山葡萄及烏蘝莓在螯合亞鐵離子的能力中效用分別有達到70%及60%以上。清除DPPH自由基試驗中翼莖葡萄最有效,清除率高達95%以上。抗肝脂質過氧化試驗中以廣東山葡萄最好,其抑制率有95%以上。白蘝對於抗氧化雖然無特別效用,但在人類乳癌細胞株MCF-7毒殺試驗上有很好的成效其抑制細胞生長率為95%以上。在全部實驗方面,山葡萄及烏蘝莓的葉部位都具有很好的抗氧化能力及毒殺乳癌細胞株功效;山葡萄的葉在MCF-7細胞毒殺試驗中的抑制率為82%。本論文希望藉由六種葡萄科植物的抗氧化能力來達到預防癌症發生及治療乳癌的輔助藥材。
英文摘要 The Vitaceae is kind of material for cleaning-heat in medicine. Normally, we use it for anti-flammatory, treating for liver trouble or trauma dressing. In this paper, we will take some familiar Vitaceae plants to test for anti-oxidation and killing human breast cancer cells MCF-7.
In this experiment, we use different kind of solvent to extract Vitaceae plants to test of antioxidae, includes clean DPPH free radical, ferrous ions chelating effects, restrain producing superoxide anion, ferric reducing ability of plasma and anti-liver lipid peroxidation assay.
In result of DPPH free radical clean test, we got good effect that using 50% ethanol and ethanol extracted Vitaceae plants in the test of ferrous ion chelating effects have over 50% restraint rate by using 50% ethanol. But it non-effect about restraining superoxide ions and cleaning. In this project for 47 sample. It showed that it has better effect by using 50% ethanol to be solvent to extract Vitaceae plants than other solvents. In antioxidant experimental, Vitis thunbergi and Cayratia japonica in ferrous ions chelating have good effect, it was 70% and 60% respectively. The assay of DPPH free radical clean test best is Cissus pteroclada, it has over 95%, in anti-liver lipid peroxidation assay, the best is Ampelopsis cantoniensis, it has over 95% too, and Ampelopsis Japonica has well effect on killing human breast cancer cells MCF-7 but have no effect about anti-oxidation.
All of experimental, we found the leave of Ampelopsis revipedunculata and Cayratia japonica have good effect of antioxidation and anti-breast cancer cell, Ampelopsis revipedunculata leave inhibit human breast cancer cell MCF-7 effect reach over 82%.
In this paper, we hope go the way by use antioxidantion to prevention cancer and treatment breast cancer adjuvant medicines with six kind of Vitaceae plants.
論文目次 目錄 II
縮寫表 IV
中文摘要 V
英文摘要 VII

一、前言 1
活性氧與自由基簡介 3
乳癌簡介 5
葡萄科植物簡介 6
ROS跟乳癌 8
二、實驗材料 10
(一)葡萄科植物 10
(二)細胞株 10
(三)實驗動物 10
(四)試藥 10
(五)儀器 12
三、實驗方法 13
(一)藥材之萃取 13
(二) 抗自由基試驗 13
(三) MCF-7細胞毒性試驗 20
四、實驗結果 23
(一)藥材萃取 23
(二)抗氧化試驗 23
1.清除DPPH自由基 23
2.清除超氧陰離子 23
3.螯合亞鐵離子 24
4.還原三價鐵離子能力測定 24
5.抗肝脂質過氧化 25
(三)MCF-7乳癌細胞株 26
五、討論 27
結論 35
圖表 36
參考文獻 53
參考文獻 1. Choliparambil K.P. et al. 1998 Study on lipid peroxidation potential in different tissues induced by ascorbate-Fe2+: Possible factors involved in their differential susceptibility. Molecular and Cellular Biochemistry 178: 197-202.
2. Cheeseman K.H. et al. 1993 An introduction to free radical biochemistry. British Medical Bulletin 49: 481-493.
3. Grace P. A. 1994 Ischaemia-reperfusion injury. Br. J. Surg. 81: 637-647
4. Grace P. A. 1994 Ischaemia-reperfusion injury. Br. J. Surg. 81: 637-647
5. Francisco B. M. et al. 1996 Lipid peroxidation prodicts in human subretinal fluid. Free rad. Biol. And Med. 20: 899-930.
6. Blount S, Griffiths HR, Lunec J. 1989 Reactive oxygen species induce antigenic changes in DNA. FEBS letters.Mar 13;245:100-104.
7. Stadtman ER, Levine RL. 2003 Free radical-mediated oxidation of free amino acids and amino acid residues in proteins. Amino Acids. Dec;25:207-218.
8. Johar D. et al. 2004 Inflammatory response, reactive oxygen species, programmed (necrotic-like and apoptotic) cell death and cancer. Roczniki Akademii Medycznej Bialymstoku 49: 31-39.
9. Evans P. et al. 1999 Free radical and hearing. Cause, consequence, and criteria. Ann N Y A Cad Sci. 884: 19-40.
10. Steinberg D. 1992 Antioxidants in the prevention of human atherosclerosis. Summary of the proceedings of a National Heart, Lung, and Blood Institute Workshop: September 5-6, 1991, Bethesda, Maryland. Circulation. 85: 2338-2344.
11. Basher S. et al. 1993 Oxidative DNA damage and cellular sensitivity to oxidative stress in human autoimmune disease. Ann. Rheum. Dis. 52: 659-666.
12. Jenner P. 1994 Oxidative damage in neurodegenerative disease. Lancet. 344: 796-798.
13. Cerutti P.A. 1985 Prooxidant states and tumor promotion. Science 227: 375-381.
14. Szatrowski T. P. et al. 1991 Production of large amounts of hydrogen peroxide by human tumor cells. Cancer Res. 51: 794-798.
15. Stadtman E.R. 1992 Protein oxidation and aging. Science 257: 1220-1224
16. Cheeseman K.H. et al. 1993 An introduction to free radical biochemistry. British Medical Bulletin 49: 481-493.
17. Evans P. et al. 1999 Free radical and hearing. Cause, consequence, and criteria. Ann N Y A Cad Sci. 884: 19-40.
18. Wu D. et al. 2003 Alcohol, oxidative stress, and free radical damage. Alcohol research & health 27: 277-284.
19. Morrissey P. A. et al. 1998 Dietary antioxidants in health and disease. Int. Dairy J. 8: 463-472.
20. Niki E. 1991 Action of ascorbic acid as a scavenger of active
and stable oxygen radicals. Ame. J. of Clin. Nutr. 54: 1119-1124.
21. Squier T. C. 2001 Oxidative stress and protein aggregation during biological aging. Experimental Gerontology 36: 1539-1550.
22. Lee H. et al. 2004 Protective effect of green tea polyphenol EGCG against neuronal damage and brain edema after unilateral cerebral ischemia in gerbils. Journal of Neuroscience Research 77: 892-900.
23. Wu D. et al., 2003 Alcohol, oxidative stress, and free radical damage. Alcohol research & health 27: 277-284.
24. 呂素英。2002。中醫護理學,知音出版社,206-207。
25. 吳素珍。1990。當代婦科護理學,南山堂。
26.甘偉松。1969。藥用植物學,國立中國醫藥研究所,373-375。
27. Wu MJ, Yen JH, Wang L, Weng CY. 2004. Antioxidant activity of Porcelainberry (Ampelopsis brevipedunculata (Maxim.) Trautv.) Am J Chin Med. 32:681-693.
28. Yabe N, Matsui H. 2000. Ampelopsis brevipedunculata (Vitaceae) extract inhibits a progression of carbon tetrachloride-induced hepatic injury in the mice. Phytomedicine. Dec;7:493-498.
29. Tan TW, Tsai HY, Chen YF, Chung JG. 2004 Induction of apoptosis in human promyelocytic leukemia HL-60 cells by Ampelopsis cantoniensis crude extract. In Vivo. Jul-Aug;18:457-462.
30. Lee YL, Lee MH, Chang HJ, Huang PY, Huang IJ, Cheng KT, Leu SJ. 2009 Taiwanese native plants inhibit matrix metalloproteinase-9 activity after ultraviolet B irradiation.Molecules. Mar 6;14:1062-1071.
31. Ku KT, Huang YL, Huang YJ, Chiou WF. 2008. Miyabenol A inhibits LPS-induced NO production via IKK/IkappaB inactivation in RAW 264.7 macrophages: possible involvement of the p38 and PI3K pathways. J Agric Food Chem. Oct 8;56:8911-8918.
32. 曾智應。2005。粉藤莖部化學成分與生物活性之研究。屏東科技大學生物科技研究所碩士論文
33. Alshawsh MA, Mothana RA, Al-Shamahy HA, Alsllami SF, Lindequist U. 2007 Assessment of antimalarial activity against Plasmodium falciparum and phytochemical screening of some Yemeni medicinal plants. Evid Based Complement Alternat Med. Oct 22.
34. Kim JH, Ju EM, Lee DK, Hwang HJ. 2002. Induction of apoptosis by momordin I in promyelocytic leukemia (HL-60) cells. Anticancer Res. May-Jun;22:1885-1889.
35. Szatrowski TP, Nathan CF. 1991,Production of large amounts of hydrogen peroxide by human tumor cells. Cancer Res 51:794-798.
36. Toyokuni S, Okamoto K, Yodoi J, Hiai H. 1995. Persistent oxidative stress in cancer. FEBS Lett 358:1-3.
37. Sipe HJ Jr, Jordan SJ, Hanna PM, Mason RP. 1994. The metabolism of 17 beta-estradiol by lactoperoxidase: a possible source of oxidative stress in breast cancer. Carcinogenesis, 15: 2637-2643
38. Wiseman H, Halliwell B. 1996. Damage to DNA by reactive oxygen and nitrogen species: role in inflammatory disease and progression to cancer. Biochem J, 313:17-29.
39. Shimada, K., Fujikawa, K., Yahara, K. and Nakamura, T. 1992. Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. J. Agric. Food Chem. 40: 945-948.
40. Robak J, Gryglewski RJ. 1988 Flavonoids are scavengers of superoxide anions. Biochem Pharmacol. Mar 1;37:837-841.
41. Dinis TC, Maderia VM, Almeida LM. 1994 Action of phenolic derivatives (acetaminophen, salicylate, and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Arch Biochem Biophys. Nov 15;315:161-169.
42. Iris F. F. Benzie and J. J. Strain . 1996. The Ferric Reducing Ability of Plasma (FRAP) as a Measure of “Antioxidant Power”: The FRAP Assay. ANALYTICAL BIOCHEMISTRY 239, 70–76
43. 楊松峻。2005。細本山葡萄(Vitis thunbergii SIEB. et ZUCC.)莖部化學成分之研究。中國醫藥大學藥學研究所碩士論文。
44. Gauthier C, Legault J, Piochon M, Lavoie S, Tremblay S, Pichette A. 2009. Synthesis, cytotoxicity, and haemolytic activity of chacotrioside lupane-type neosaponins and their germanicane-type rearrangement products.Bioorg Med Chem Lett. Apr 15;19:2310-2314.
45. Saleem M, Maddodi N, Abu Zaid M, Khan N, bin Hafeez B, Asim M, Suh Y, Yun JM, Setaluri V, Mukhtar H. 2008. Lupeol inhibits growth of highly aggressive human metastatic melanoma cells in vitro and in vivo by inducing apoptosis. Clin Cancer Res. Apr 1;14:2119-2127.
46. Mullauer FB, Kessler JH, Medema JP. 2009. Betulin is a potent anti-tumor agent that is enhanced by cholesterol. PLoS One. Apr 28.
47. Fulda S, Kroemer G. 2009. Targeting mitochondrial apoptosis by betulinic acid in human cancers. Drug discovery today. Jun 9.
48. Pyo JS, Roh SH, Kim DK, Lee JG, Lee YY, Hong SS, Kwon SW, Park JH. 2009. Anti-cancer effect of Betulin on a human lung cancer cell line: a pharmacoproteomic approach using 2 D SDS PAGE coupled with nano-HPLC tandem Mass Spectrometry. Planta Med. Feb;75:127-131.
49. 王文玲。2002。Apigenin, 17α-estradiol 和flavone引起人類胃癌細胞株(SC-M1)生長的抑制和調節細胞週期相關基因的表現。中國醫藥學院醫學研究所碩士論文。
50. 黃淑于。1992。Apigenin對TPA在NIH3T3細胞誘發c-jun及c-fos 原致癌基因表現影響之探討。國立台灣大學毒理學研究所碩士論文。
51. 陳鍵萱。2002。 木犀草素誘發人類肝癌細胞凋亡。高雄醫學大學天然藥物研究所碩士論文。
52. 楊芳瑄。2003。槲黃素對造骨細胞活性之影響。中原大學醫學工程研究所碩士論文。
53. Xu Z, Liu X, Xu G. 1995.Chemical constituents of roots of Ampelopsis brevipedunculata (Maxim) Trautv.Zhongguo Zhong Yao Za Zhi. Aug;20:484-486, 512.
54. 高安平。2005。南嶺前胡葉部與廣東山葡萄之成分研究。國立成功大學化學系碩士論文。
55. Choi YH, Yan GH. 2009 .Anti-allergic effects of scoparone on mast cell-mediated allergy model. Phytomedicine. Jun 11
56. 曹聖凰。1996。台灣產露兜樹科植物林投之抗氧化活性及化學成分之研究。國立中興大學化學系碩士論文。
57. Pellegrini N, Valtueña S, Ardigò D, Brighenti F, Franzini L, Del Rio D, Scazzina F, Piatti PM, Zavaroni I. 2009. Intake of the plant lignans matairesinol, secoisolariciresinol, pinoresinol, and lariciresinol in relation to vascular inflammation and endothelial dysfunction in middle age-elderly men and post-menopausal women living in Northern Italy.Nutr Metab Cardiovasc Dis. Apr 8
58. Je-Chiuan Ye. 2005. Analysis of β- sitosterol in Knoxia corymbosa. 國立嘉義大學生物藥學研究所碩士論文。
59. Jayaprakasha GK, Mandadi KK, Poulose SM, Jadegoud Y, Nagana Gowda GA, Patil BS. 2007. Inhibition of colon cancer cell growth and antioxidant activity of bioactive compounds from Poncirus trifoliata (L.) Raf. Bioorg Med Chem. Jul 15;15(14):4923-32.
60. 張均嵐。1996。1-nor-3-dehydrobetulinic acid, 1-deformylzizyberenalic acid和1-decarboxyceanothic acid 對於人類卵巢腺癌細胞株毒性之探討。國立陽明大學生物藥學研究所碩士論文。
61. Noungoue DT, Chaabi M, Ngouela S, Antheaume C, Gut J, Rosenthal PJ, Lobstein A, Tsamo E. 2009. Antimalarial compounds from the stem bark of Vismia laurentii. Zeitschrift für Naturforschung. C, Journal of biosciences. Mar-Apr;64:210-214.
62. Yun Y, Han S, Park E, Yim D, Lee S, Lee CK, Cho K, Kim K. 2003. Immunomodulatory activity of betulinic acid by producing pro-inflammatory cytokines and activation of macrophages. Archives of pharmacal research. Dec;26:1087-1095.
63. Yasunari Takada, and Bharat B. Aggarwal 2003. Betulinic Acid Suppresses Carcinogen-Induced NF-κB Activation Through Inhibition of IκBα Kinase and p65 Phosphorylation:Abrogation of Cyclooxygenase-2 and Matrix Metalloprotease-9. Journal of Immunology, 171: 3278–3286.
64. Chi-Yun Cheng. 2006. Anti-cancer mechaniams of ursolic acid:characterization of cell cycle arrest and apoptotic machinery.高雄醫學大學天然藥物研究所碩士論文。
65. Chi-Ling Wu. 2006. Ursolic acid inhibits the proliferation and metastasis of cancer cells in vivo and in vitro. 國立成功大學藥理學研究所碩士論文。
66. 曾智應。2004。粉藤莖部化學成分與生物活性之研究。屏東科技大學生物科技研究所碩士論文。
67. Akihisa T, Ogihara J, Kato J, Yasukawa K, Ukiya M, Yamanouchi S, Oishi K. 2001. Inhibitory effects of triterpenoids and sterols on human immunodeficiency virus-1 reverse transcriptase. Lipids. May;36:507-512.
68. Sewram V, Raynor MW, Mulholland DA, Raidoo DM. 2000 The uterotonic activity of compounds isolated from the supercritical fluid extract of Ekebergia capensis.Journal of pharmaceutical and biomedical analysis. Dec;24:133-145.
69. Abbas FA, Al-Massarany SM, Khan S, Al-Howiriny TA, Mossa JS, Abourashed EA. 2007. Phytochemical and biological studies on Saudi Commiphora opobalsamum L. Natural product research. May;21:383-391.
70. Guo L ibin, L u Y an, Chen S hu ip . 1996. Study on The Chemical Constituents of Ampelopsis Japonica (Thunb. ) Makino.廣東藥學院學報(ACAD J GCP) 12: 145-147
71. Noa M, Mendoza S, Más R. 2005 Effect of D-003 on intimal thickening and circulating endothelial cells in rabbit cuffed carotid artery. Journal of medicinal food. Summer;8:237-241.
72. Lee DK, Kim B, Lee SG, et al. 1998. Momordins inhibit both AP-1 function and cell proliferation . Anticancer Res, 18: 119-124.
73. Park S, Lee DK, Whang YH, et al. 2000. Momordin I, a compound of ampelopsis radix, inhibits AP-1 activation induced by phorbolester. Cancer Letter 152: 1-8.
74. Han XH, Hong SS, Hwang JS, Lee MK, Hwang BY, Ro JS. 2007 Monoamine oxidase inhibitory components from Cayratia japonica Archives of pharmacal research.Jan;30:13-17.
75. Bradford LW. 1976 Problems of ethics and behavior in the forensic sciences. Journal of forensic sciences Oct 21:763-768.

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系統識別號 U0007-2407200911324700
論文名稱(中文) 大安水蓑衣成分及相關活性探討
論文名稱(英文) The constituents and their related activities of Hygrophila pogonocalyx Hayata
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 97
學期 2
出版年 98
研究生(中文) 林建宏
學號 M303096007
學位類別 碩士
語文別 中文
口試日期 2009-07-16
論文頁數 152頁
口試委員 指導教授-李美賢
共同指導教授-李宗徽
委員-郭悅雄
委員-張芳榮
委員-張溫良
關鍵字(中) 大安水蓑衣
臺灣特有種
神經細胞保護
人類黑色素細胞
酪氨酸酵素
關鍵字(英) Hygrophila pogonocalyx Hayata
endemic species in Taiwan
nerve growth factor-differentiated PC12 cell
human epidermal melanocytes
學科別分類
中文摘要 大安水蓑衣(Hygrophila pogonocalyx Hayata)為爵床科水蓑衣屬(Hygrophila)多年生水生挺水植物,為台灣特有種,產於台中縣臨海鄉鎮。根據本實驗室先前實驗結果顯示,大安水蓑衣95%乙醇萃取物,具清除自由基的活性(HO•、O2•‾及ABTS•‾,IC50 = 0.78、12.75及 7.94 ug/mL),因其被國際自然及自然資源保育聯盟 (IUCN),列為瀕臨絕滅級 (endangered ),故本研究將組織培養的大安水蓑衣地上部以 95% 乙醇萃取,並進行成分分離及其活性探討。經管柱層析分離純化得到十三個化合物,由物理和光譜相關數據解析後,分別為flavones 結構 luteolin 7-O-β-D-glucopyranoside(1),luteolin 7-O-β- D-glucuronide(2);flavonols 結構 quercetin(3),isoquercitrin(4),rutin(5),myricetrin(6);phenylethanoid glycosides 結構 acteoside(7),isoacteoside(8),β-ethoxylacteoside(9);alkylated glycoside 結構 3-O-[β-D-apiofuranosyl-(1→6)-β-D-glucopyranosyl]oct-1-en-3-ol(10),3-O-[β-L-xylopyranosyl-(1→6)-β-D-glucopyranosyl]oct-1-en-3-ol(11);及 steroid 結構 β-sitosterol(12)及 stigmasterol(13),以上十三個化合物均首次由大安水蓑衣中分離得到。此十三個化合物,除化合物 3 及 4 因量少、化合物 12 及 13 因為混合物未測試外,化合物1、2、5 ~ 11 進行神經生長因子分化的 PC12 細胞活性篩選試驗,在細胞存活率測試中,於100 μM 濃度下,發現九種化合物的細胞存活率皆大於 85%,進一步進行以六-羥基多巴胺誘導神經生長因子分化的 PC12 細胞毒性模式,結果顯示化合物 luteolin 7-O-β-D-glucopyranoside(1)具最佳細胞保護活性。十三個化合物,除化合物 12 及 13 因為混合物未測試外,化合物 1 ~ 11 進行人類黑色素細胞內酪氨酸酵素活性試驗,在細胞存活率測試中,於100 μM 濃度下,發現十一種化合物的細胞存活率皆大於 85%,進一步對人類黑色素細胞內酪氨酸抑制活性測試,結果顯示化合物 acteoside(7) 具有較佳抑制酪氨酸酵素活性效果。
英文摘要 Hygrophila pogonocalyx Hayata is an endemic species in Taiwan. It was found along the seashore in Taichung country and was endangered according to the Red List Categories of the International Union for Conservation of Nature and Natural Resources (IUCN). Our previous results showed that the 95% ethanol extract of Hygrophila pogonocalyx exhibited free radical scavenging activities (HO•, O2•‾ and ABTS•‾, IC50 = 0.78, 12.75 and 7.94 ?慊/mL). Therefore, the active constituents of H.pogonocalyx from tissue cultures (providing by Dr. Chin-Wen Ho, Tatung University, Taipei) are isolated in the present study. The structures of isolated compounds were identified by various physical and spectroscopic characterizations (eg. ESI-MS, UV, IR and NMR etc.). Thirteen compounds from Hygrophila pogonocalyx Hayata were identified in this study, including two flavones, luteolin 7-O-β-D-glucopyranoside (1) and luteolin 7- O-β-D-glucuronide (2); four flavonols, quercetin (3), isoquercitrin (4), rutin (5) and myricetrin (6); three phenylethanoid glycosides, acteoside (7), isoacteoside (8) and β-ethoxyl-acteoside (9); two alkylated glycosides, 3-O-[β-D- apiofuranosyl-(1→6)-β-D-glucopyranosyl]oct-1-en-3-ol (10) and 3-O- [β-L-xylo- pyranosyl-(1→6)-β-D-glucopyranosyl]oct-1-en-3-ol (11) and two steroid, β-sitosterol (12) and stigmasterol (13). Those 9 compounds were further evaluated the neurocytoprotective activity in NGF-differentiated PC12 cells, and compound 1 shows the most potent activity. Those 11 compounds were also evaluated the inhibition of cellular tyrosinase activities in human epidermal melanocytes(HEMn). Cell viabilities of compounds 1 ~ 11 were higher than 85%, the compound 7 was the most potent in inhibiting tyrosinase activity.
論文目次 縮寫表………………………………………………………………XI
中文要………………………………………………………………XII
英文摘要……………………………………………………………XIV
壹、緒論
一、大安水蓑衣(Hygrophila pogonocalyx Hayata)
基本介紹及其同屬植物研究回顧
(一)大安水蓑衣(Hygrophila pogonocalyx Hayata)
基本介紹……………………………………………………………1
(二)水蓑衣屬(Hygrophila spp.)植物相關介紹……2
(三)水蓑衣屬(Hygrophila spp.)植物近代藥理活性
研究回顧……………………………………………………………5
(四)水蓑衣屬(Hygrophila spp.)植物之化學成份
研究回顧……………………………………………………………7
二、PC12 細胞株及相關介紹
(一)PC12 細胞株簡介……………………………………11
(二)6-羧基多巴胺(6-hydroxydopamine, 6-OHDA)
之應用…………………………………………………11
三、人類黑色素細胞與其相關介紹
(一)人類黑色素細胞(Melanocyte)………………………13
(二)黑色素(Melanin)……………………………………13
(三)黑色素的生成(Melanogenesis)……………………14
(四)抑制黑色素生成之機制………………………………14
四、研究動機…………………………………………………16
貳、實驗材料與方法
一、材料及儀器
(一)一般試藥及溶媒………………………………………17
(二)色層分析法材料………………………………………17
(三)儀器……………………………………………………18
二、大安水蓑衣(Hygrophila pogonocalyx Hayata)活性成分
分離流程…………………………………………………20
三、水解和單糖的組成分析
1. 高效陰離子交換層析法………………………………23
2. 薄層層析法……………………………………………24
四、抑制酪氨酸酵素實驗
(一)人類黑色素細胞培養…………………………………25
(二)人類黑色素細胞存活率試驗…………………………25
(三)蛋白定量法……………………………………………26
(四)人類黑色素細胞內酪氨酸酵素活性試驗……………27
五、神經保護實驗
(一)PC 12 細胞培養………………………………………28
(二)PC 12 細胞存活率試驗………………………………28
(三)PC12細胞受6-OHDA毒害及活性成分保護之實驗……29
六、各成分之物理數據………………………………………30
叁、結果與討論
一、大安水蓑衣(Hygrophila pogonocalyx Hayata)之化合物
其結構鑑定………………………………………………44
二、大安水蓑衣成分之活性探討……………………………79
肆、結論……………………………………………………………83
伍、參考文獻………………………………………………………87
參考文獻 1. 蔡紹斌. 清水地誌系列之三 清水鎮植物誌. 1999:8.
2. 黃朝慶;文紀鑾. 台灣野生植物資料庫特有生物保育中心保育季刊 1999;26:4.
3. 行政院農委會林務局自然資源與生態資料庫.
4. 楊遠波;劉和義;彭鏡毅;施炳霖;呂勝由. 台灣維管束植物簡誌. 1998;第四卷.
5. Vijayakumar M, Govindarajan R, Shirwaikar A KV, Rawat A, Kumar S, Mehrotra S, Pushpangadan P. Free radical scavenging and lipid peroxidation inhibition potential of Hygrophila auriculata. Natural Products Sciences 2005;11(1):22-26.
6. Shanmugasundaram P, Venkataraman S. Hepatoprotective and antioxidant effects of Hygrophila auriculata (K. Schum) Heine Acanthaceae root extract. Journal of Ethnopharmacol 2006;104(1-2):124-128.
7. Mazumdar U, Gupta M, Maiti S, Mukherjee D. Antitumor activity of Hygrophila spinosa on Ehrlich ascites carcinoma and sarcoma-180 induced mice. Indian Journal of Experimental Biology 1997;35(5):473-477.
8. Mazumdar U, Gupta M, Maiti S. Effect of petroleum ether extract from Hygrophila spinosa on hematological parameters and hepatorenal functions in mice. Indian Journal of Experimental Biology 1996;34(12):1201-1203.
9. Khan MR, Omoloso AD. Antibacterial activity of Hygrophila stricta and Peperomia pellucida. Fitoterapia 2002;73(3):251-254.
10. Ariza SY, Rueda DC, Rincon V J, Linares EL, Guerrero MF. Pharmacological effects on the central nervous system induced by coumarin, isolated from Hygrophila tyttha Leonard. Vitae 2007;14(2):51-58.
11. Haq QN, Nabi MN. Studies on oil from the seeds of Hygrophila spinosa. Bangladesh Journal of Science Industrial Research. 1978;13(1-4):29-32.
12. Jensen SR, Nielsen BJ. Hygrophiloside, an iridoid glucoside from Hygrophila difformis (Acanthaceae). Phytochemistry 1985;24(3):602-603.
13. Daniel M. medicinal plants: chemistry and properties. 2005:192-193.
14. Greene L, Tischler, AS. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proceedings of the National Academy of Sciences of the United States of America 1976;73(7):2424-2428.
15. Heneka M, Loschmann P, Gleichmann M, Weller M, Schulz JB, Wullner U, Klockgether T. Induction of Nitric Oxide Synthase and Nitric Oxide-Mediated Apoptosis in Neuronal PC12 Cells After Stimulation with Tumor Necrosis FActor-α/Lipopolysaccharide. Journal of Neurochemistry 1998;71(1):88-94.
16. Huang Y, Chang, AY, Huang, CM, Huang, SW, Chan, SH. . Proteomic analysis of lipopolysaccharide-induced apoptosis in PC12 cells. Proteomics 2002;2(9):1220-1228.
17. Kamata H, Tanaka C, Yagisawa H, Matsuda S, Gotoh, Y, Nishida, E, Hirata, H. Suppression of nerve growth factor-induced neuronal differentiation of PC12 cells. N-acetylcysteine uncouples the signal transduction from ras to the mitogen-activated protein kinase cascade. The Journal of biological chemistry 1996;271(51):33018-33025.
18. Leppa S SR, Ansorge W, Bohmann D. Differential regulation of c-Jun by ERK and JNK during PC12 cell differentiation. The EMBO Journal 1998;17(15):4404-4413.
19. Morooka T, Nishida E. Requirement of p38 Mitogen-activated Protein Kinase for Neuronal Differentiation in PC12 Cells. The Journal of biological chemistry 1998;273(38):24285-24288.
20. Obin M, Eugene X, Arthur H, James J, Allen T. Neurite Outgrowth in PC12 Cells. The Journal of biological chemistry 1999;274(17):11789-11795.
21. Richard MK, David MJ. Pharmacological actions of 6-hydroxydopamine. Pharmacological reviews 1974;26(3):199-288.
22. Yoshiro S, Keiko N, Yoko O, Tomoya, K, Yasukazu, Y, Keiko, N. Molecular mechanisms of 6-hydroxydopamine-induced cytotoxicity in PC12 cells: Involvement of hydrogen peroxide-dependent and -independent action. Free Radical Biology & Medicine 2007;42: 675-685.
23. Mayo JC, Sainz RM, Antolin I, Rodringuez, C. Ultrastructural confirmation of neuronal protection by melatonin against the neurotoxin 6-hydroxydopamine cell damage. Brain Research 1999;818(2):221-227.
24. Junqueira LC, Carneiro J, Kelley RO. 基礎組織學. 合記圖書出版社 2000.
25. Walter C, Quevedo JR. The control of color in mammals. Am Zool 1969;9(2):531-540.
26. Prota G. Recent advances in the chemistry of melanogenesis in mammals. Journal of Investgative Dermatology 1980;75(1):122-127.
27. Pawelek JM, Korner AM. The biosynthesis of mammalian melanin. Am Sciences 1982;70(2):136-145.
28. Marmol V, Beermann F. Tyrosinase and related proteins in mammalian pigmentation. Federation of European Biochemical Societies Letters 1996;381(3):165-168.
29. Schallreuter KU. Advances in melanocyte basic science research. Dermatologic Clinics 2007;25(3):283-291.
30. Lerner A, Fitzpatrick T, Calkins E. Mammalian tyrosinase; preparation and properties. Journal of Biological Chemistry 1949;178(1):185-195.
31. Korner A, Pawelek J. Mammalian tyrosinase catalyzes three reactions in the biosynthesis of melanin. Science 1982;217(4565):1163-1165.
32. Mishima Y, Hatta S, Ohyama Y, Inazu, M. Induction of melanogenesis suppression: cellular pharmacology and mode of differential action. Pigment Cell Research 1988;1(6):367-74.
33. Land EJ, Ramsden CA, Riley PA. Tyrosinase autoactivation and the chemistry of ortho-quinone amines. Accounts of Chemical Research 2003;36(5):300-308.
34. Maeda K, Fukuda M. Arbutin: mechanism of its depigmenting action in human melanocyte culture. Journal of Pharmacology Experimental Therapeutics 1996;276(2):765-769.
35. Mishima Y HS, Ohyama Y, Inazu M. induction of melanogenesis suppression: cellular pharmacology and mode of differential action. Pigment Cell Res 1988;1(16):367-74.
36. Kumano Y, Sakamoto T, Iwai I, Tanaka, M, Tamamoto, I. In vitro and in vivo prolonged biological activities of novel vitamin C derivative, 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G), in cosmetic fields. Journal of Nutritional Science and Vitaminology 1998;44(3):345-359.
37. Funasaka Y, Chakraborty AK, Komoto M, Ohashi, A, Ichihashi, M. The depigmenting effect of alpha-tocopheryl ferulate on human melanoma cells. British Journal of Dermatology 1999;141(1):20-29.
38. Tachibana M, Takeda K, Nobukuni Y. Ectopic expression of MITF, a gene for waardenburg syndrom type 2, converts fibroblasts to cells with melanocyte characteristics. Nature Genetics 1996;14(1):50-54.
39. Huang JC, Wang WK, Hong KH, Chiang TY. Population differentiation and phylogeography of Hygrophila pogonocalyx based on RAPD fingerprints. Aquatic Botany 2001;70(4):269-280.
40. Huang JC, Wang WK, Peng CI, Chiang, T Y. Phylogeography and conservation genetics of Hygrophila pogonocalyx (Acanthaceae) based on atpB-rbcL noncoding spacer cpDNA. Journal of Plant Research 2005;118(1):1-11.
41. Shi S, Zhao Y, Zhou H, Zhang Y, Jiang X, Huang K. Identification of antioxidants from Taraxacum mongolicum by high-performance liquid chromatography-diode array detection-radical-scavenging detection-electrospray ionization mass spectrometry and nuclear magnetic resonance experiments. Journal of Chromatography 2008;1209(1-2):145-152.
42. Lee MH, Son YK, Han YN. Tissue factor inhibitory flavonoids from the fruits of Chaenomeles sinensis. Archives of Pharmacal Research 2002;25(6):842-850.
43. Hsieha TJ, Wu YC, Chena CY. Chemical Con stituents from the Leaves of Epimedium sagittatum. The Chinese Pharmaceutical Journal(Taipei, Taiwan) 2003(55):121-128.
44. Lu Y, Foo LY. Identification and quantification of major polyphenols in apple pomace. Food Chemistry 1997;59(2):187-194.
45. Kazuma K, Noda N, Suzuki M. Malonylated flavonol glycosides from the petals of Clitoria ternatea. Phytochemistry (Elsevier) 2003;62(2):229-237.
46. Ibrahim I, Mahmoud MS, Marzouk AM, Moharrama, M R. Acylated flavonol glycosides from Eugenia jambolana leaves. Phytochemistry 2001;58(2):1239-1244.
47. Henry M, Roussel JL, Andary C. Verbascoside production in callus and suspension cultures of Hygrophila erecta. Phytochemistry 1987;26(7):1961-1963.
48. Kim HJ, Woo ER, Shin CG, Hwang DJ, Park H, Lee YS. HIV-1 integrase inhibitory phenylpropanoid glycosides from Clerodendron trichotomum. . Archives of Pharmacal Research 2001;24(6):286-291.
49. Jun W, Huang J, Long L, Huang L. Phenylethanoid and aliphatic alcohol glycosides from Acanthus ilicifolius. Phytochemistry 2003;63(4):491-495.
50. Zou J, Zhu YD, Zhao WM. Two new alkyl glycosides from Clerodendranthus spicatus. Journal of Asian Natural Products Research 2008;10(7):602-606.
51. Yamamura S, Ozawa K, Ohtani K, Kasai R, Yamasaki K. Antihistaminic flavones and aliphatic glycosides from Mentha spicata. Phytochemistry 1998;48(1):131-136.
52. Hisash K, Noriko S, Akiko H, Haruo O. Sterol glucosides from prunella vulgaris. Phytochemistry 1990;29(7):2351-2355.
53. Lemanska K, Szymusiak H, Tyrakowska B, Zielinski, R, Soffers, A E, Rietjens, I M. The influence of pH on antioxidant properties and the mechanism of antioxidant action of hydroxyflavones. Free Radical Biology & Medicine 2001;31(6):869-881.
54. Alton J, Dugas J, Jose CA, Gloria, C B, Kimberly, L P, Nikolaus, H F, Gary, W. Evaluation of the Total Peroxyl Radical-Scavenging Capacity of Flavonoids: Structure-Activity Relationships. journal of naturall products 2000;63(3):327-331.
55. Kahkonen MP, Heinonen M. Antioxidant activity of anthocyanins and their aglycons. Journal of Agricultural and Food Chemistry 2003;51(3):628-633.
56. Navindra PS, Muraleedharan GN. Inhibition of Lipid Peroxidation and Structure-Activity-Related Studies of the Dietary Constituents Anthocyanins, Anthocyanidins, and Catechins. Journal of Agricultural and Food Chemistry 2002;50(19):5308-5312.

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系統識別號 U0007-2707200912411400
論文名稱(中文) 天然物-台灣大戟抗光老化活性成分之研究
論文名稱(英文) Anti-photoaging Activity of Natural Products Research in Euphorbia formosana
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 97
學期 2
出版年 98
研究生(中文) 余佳純
學號 M303096016
學位類別 碩士
語文別 中文
口試日期 2009-06-19
論文頁數 200頁
口試委員 委員-郭悅雄
委員-吳天賞
委員-李宗徽
指導教授-李慶國
共同指導教授-蕭哲志
關鍵字(中) 抗光老化
基質金屬蛋白酶
細胞外基質
關鍵字(英) anti-photoageing
matrix metalloproteinases (MMPs)
extracellular matrix (ECM)
學科別分類
中文摘要 隨著地球臭氧層的破壞,使得皮膚曝曬在紫外線下的機會增加,導致皮膚提早老化和皮膚癌的發生;在光老化的過程中,基質金屬蛋白酶(Matrix metalloproteinase,簡稱MMPs)扮演了一個重要的角色,本研究藉由抑制 MMP-2 和 MMP-9 的活性,使皮膚細胞外基質(Extracellular matrix,簡稱ECM)不受到 MMP-2 和 MMP-9 的分解而造成皮膚老化。因此,先前本實驗室已將台灣常用的 49 種藥用植物利用 MMP-2 和 MMP-9進行抗光老化的酵素活性篩選,其中以台灣大戟(Euphorbia formosana Hayata)植物抑制 MMP-2 和 MMP-9 的效果為最佳。
將台灣大戟植物利用人類纖維肉瘤細胞(Human fibrosarcoma cell) HT-1080 細胞中的 MMP-2 與 MMP-9 的活性追蹤方式進行成分的分離及純化,並利用化合物之核磁共振與物理數據等光譜數據,共計純化出 38 個化合物,包括:九個 aliphatic 類化合物、九個 triterpenoid 類化合物、六個 diterpenoid 類化合物、三個 steroid 類化合物、二個 phaeophorbide 類化合物、一個 coumarin 類化合物、一個 aromatic 類化合物、一個 norisoprenoid 類化合物、一個 purine alkaloid 類化合物、一個 polyketide 類化合物、一個 polyprenol 類化合物、一個 lignin 類化合物、二個 others 類化合物。上述38個化合物,其中seco-Helioscopinolde (EF12)、3β,7β-Dihydroxy-ent-abieta-8,13-diene-16,12-olide (EF13) 為新化合物。
將純化合物對於 MMP-2 和 MMP-9 作活性篩選,實驗結果發現這些化合物對於 MMP-2 與 MMP-9 的抑制效果不佳,與先前的研究結果活性表現不一致,推測是先前的研究中,利用萃取液直接對 MMP-2 和 MMP-9 作酵素反應,而在本研究中則是利用化合物對 HT-1080 細胞作試藥處理,經由細胞內的訊息調控後,取其細胞外液作 Gelatin Zymography 實驗,分析化合物對 MMP-2 與 MMP-9 的活性表現,因此,針對於 MMP 的抑制作用可能藉由不同的基因表現方式,而造成兩者之間對 MMP-2 與 MMP-9 活性表現的差異,期待未來可藉由不同的化合物調控 MMP 的蛋白表現或酵素活性,期待能開發作為新一代天然的抗光老化產品。
英文摘要 With increasing UV exposure of skin leads to acute and chronic detrimental cutaneous effects, which may result in development of skin malignancies and photoaging. The Matrix Metalloproteinases (MMPs) play an important role in the photoaging process, which degrade macromolecules of the extracellular matrix (ECM) to increase the happening of wrinkling, sagging and laxity. We examine the MMP-2 and MMP-9 activity for the factor in photoaging process that can degrade collagen and components of the elastic network.
In the previous studies, our groups had screened 49 species of pharmaceutical plants in Taiwan by anti-photoaging screening model using MMP-2 and MMP-9 enzyme activity. The results demonstrated that Euphorbia formosana had better inhibition of MMPs activity in HT-1080 cells. In Euphorbia formosana, 38 compounds had been isolated and purified by using HPLC and spectroscopy for structure elucidation. There is included nine aliphatic compounds, nine tritepenoids, six diterpenoids, three steroids, two phaeophorbides, one coumarin, one aromatic structure, one norisoprnoid, one purine alkaloid, one polyketide, one poylprenol, one lignan, and the others. Among them, seco-Helioscopinolde (EF12), 3β,7β-Dihydroxy-ent-abieta-8,13-diene-16,12-olide (EF13) are discovered and identified in first time.
All purified compounds were evaluated the MMP-2 and MMP-9 acitivity in HT-1080 cells, and the results indicated that neither one showed the inhibition of MMP-2 and MMP-9 activity, which were not coincide with the previous study. We were speculating that the difference in crude extracts with MMP-2 and MMP-9 enzyme reaction and compounds were treated to human fibrosarcoma cell (HT-1080) by cell signaling regulation. We supposed that the acitivity of MMP-2 and MMP-9 may be modulated by several genes expression, which resulting in the difference between enzyme reaction and cell signaling. The development for MMP inhibition will require the different compounds in modulating MMP protein expression or enzyme activity, which will be an anti-photoageing products from natural source in new generation.
論文目次 博碩士論文授權書 ----------------------------------------- i
口試委員審查書 ------------------------------------------ ii
謝誌 --------------------------------------------------- iii
中文摘要 (Abstract in Chinese) --------------------------- v
英文摘要 (Abstract in English) ------------------------- vii
目錄 (Contents) ----------------------------------------- ix
圖目錄 (Figure Contents) ------------------------------- xii
表目錄 (Table Contents) -------------------------------- xix
縮寫表 (Abbreviations) --------------------------------- xxi
第一章 緒論 (Introduction)
第一節、 研究背景與動機 ---------------------------------- 1
1-1-1. 全球保養品市場 ------------------------------------ 1
1-1-2. 紫外線的簡介 -------------------------------------- 3
1-1-3. 防曬的方法 ---------------------------------------- 6
第二節、 光老化作用
1-2-1. 皮膚的簡介 ---------------------------------------- 9
1-2-2. 抗老化機制之研究 --------------------------------- 13
第三節、 基質金屬蛋白酶的簡介 --------------------------- 16
第四節、 植物的分佈及型態 ------------------------------- 27
第五節、 文獻回顧 --------------------------------------- 31
第二章 實驗結果 (Results)
第一節、 植物分配萃取與成分分離流程 --------------------- 37
第二節、 化合物之結構解析
2-2-1. Diterpenoids 之結構解析
2-2-1-1. Helioscopinolide B (EF9) ----------------------- 48
2-2-1-2. Helioscopinolide A (EF10) ---------------------- 56
2-2-1-3. Helioscopinolide C (EF11) ---------------------- 62
2-2-1-4. seco-Helioscopinolide (EF12)* ------------------ 68
2-2-1-5. 3β,7β-Dihydroxy-ent-abieta-8,13-diene-16,12-olide (EF13)* ------------------------------------------- 77
2-2-1-6. ent-(5β,8a,9β,10a,12a)-12-hydroxyatis-16-ene-
3,14-dione (EF14) --------------------------------------- 86
2-2-2. Triterpenes之結構解析
2-2-2-1. Lupenone (EF16) -------------------------------- 94
2-2-2-2. Glutinol (EF17) -------------------------------- 99
2-2-2-3. Acetylglutinol (EF18) ------------------------- 105
2-2-2-4. (-)-Euphol (EF19) ---------------------------- 111
2-2-2-5. (+)-Tirucallol (EF20) ------------------------ 119
2-2-2-6. (20R,23E)-Eupha-8,23-diene-3β-25-diol (EF21)-- 124
2-2-2-7. (20R)-Eupha-8,24-diene-3-one (EF22) ----------- 132
2-2-2-8. (20R)-Eupha-8,25-diene-3β,24??-diol (EF23)---- 136
第三節、 化合物之物理和光譜數據總整理------------------- 141
第三章 實驗材料與方法 (Materials and Methods)
第一節、 活性實驗部分
3-1-1. 實驗藥品試劑 ------------------------------------ 156
3-1-2. 實驗設備 ---------------------------------------- 158
3-1-3. 實驗方法
3-1-3-1. 細胞培養 (Cell Culture) ----------------------- 159
3-1-3-2. 細胞存活試驗 (MTT cell viability assay) ------- 160
3-1-3-3. 電泳酵素分析法 (Zymography) ------------------- 161
3-1-3-4. 統計分析 (Statistical analysis) --------------- 162
第二節、 萃取與分離純化部分
3-2-1. 分析儀器 ---------------------------------------- 163
3-2-2. 溶媒試劑 ---------------------------------------- 165
3-2-3. 氘化溶媒 ---------------------------------------- 165
3-2-4. 實驗方法 ---------------------------------------- 166
第四章 討論 (Discussion) ------------------------------ 179
第五章 參考文獻 (References) -------------------------- 182
附錄 (Appendix) ---------------------------------------- 198
參考文獻 1.鄭晴文, 日本抗老化市場發展現況及趨勢. 工業技術研究院 2008.
2.羅淑慧, 藥妝品之發展趨勢. ITIS 2003, 第四章.
3.李美賢, 中草藥功能性化妝品之應用開發. Yearbook of Chinese Medicine and Pharmacy 2007, 25, (1), 147-180.
4.Roy, C. R.; Gies, H. P.; Lugg, D. J.; Toomey, S.; Tomlinson, D. W., The measurement of solar ultraviolet radiation. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 1998, 422, (1), 7-14.
5.Goihman-Yahr, M., Skin aging and photoaging: an outlook. Clinical Dermatology 1996, 14, (2), 153-160.
6.Zhang, G.; Luo, X.; Sumithran, E.; Pua, V.; Barnetson, R.; Halliday, G.; Khachigian, L., Squamous cell carcinoma growth in mice and in culture is regulated by c-Jun and its control of matrix metalloproteinase-2 and -9 expression. Oncogene 2006, 25, 7260-7266.
7.Stierner, U., Melanocytes, moles and melanoma: a study on UV effects. Acta dermato-venereologica. Supplementum 1991, 168, 1-31.
8.Urbach, F., The historical aspects of sunscreens. Journal of Photochemistry and Photobiology B: Biology 2001, 64, 99-104.
9.Lowe, N. J., An Overview of Ultraviolet Radiation, Sunscreens, and Photo-Induced Dermatoses. Dermatologic Clinics 2006, 24, (1), 9-17.
10.Rosen, C. F., Topical and systemic photoprotection. Dermatologic Therapy 2003, 15, (1), 8-15.
11.Scharffetter-Kochanek, K.; Brenneisen, P.; Wenk, J.; Herrmann, G.; Weijan Ma, L. K.; Meewes, C.; Wlaschek, M., Photoaging of the skin from phenotype to mechanisms. Experimental Gerontology 2000, 35, (3), 307-316.
12.Gilchrest, B., Skin aging and photoaging: an overview. Journal of the American Academy of Dermatology 1989, 21, (3 Pt 2), 610-613.
13.Rittié, L.; Fisher, G. J., UV-light-induced signal cascades and skin aging. Ageing Research Reviews 2002, 1, (4), 705-720.
14.Griffiths, C. E. M., The role of retinoids in the prevention and repair of aged and photoaged skin. Clinical and Experimental Dermatology 2001, 26, (7), 613-618.
15.Fisher, G. J.; Kang, S.; Varani, J.; Bata-Csorgo, Z.; Wan, Y.; Datta, S.; Voorhees, J. J., Mechanisms of photoaging and chronological skin aging. Archives of Dermatology 2002, 138, (11), 1462-1470.
16.Trautinger, F., Mechanisms of photodamage of the skin and its functional consequences for skin ageing. Clinical and Experimental Dermatology 2001, 26, (7), 573-577.
17.Emerit, I., Free radicals and aging of the skin. EXS 1992, 62, 328-341.
18.Anthony V. Benedetto, D., The environment and skin aging. Clinics in Dermatology 1998, 16, (1), 129-139.
19.Pugliese, P. T., The skin's antioxidant systems. Dermatology Nursing 1998, 10, (6), 401-416.
20.Pugliese, P., The skin, free radicals, and oxidative stress. Dermatology Nursing 1995, 7, (6), 361-369.
21.Chung, J. H.; Seo, J. Y.; Choi, H. R.; Lee, M. K.; Youn, C. S.; Gi-eun Rhie; Cho, K. H.; Kim, K. H.; Park, K. C.; Eun, H. C., Modulation of Skin Collagen Metabolism in aged and photoaged human skin in vivo. The Journal of Investigative Dermatology 2001, 117, (5), 1218-1224.
22.Rabe, J. H.; Mamelak, A. J.; McElgunn, P. J. S.; Morison, W. L.; Sauder, D. N., Photoaging: Mechanisms and repair. Journal of the American Academy of Dermatology 2006, 55, (1), 20-21.
23.Philips, N.; Smith, J.; Keller, T.; Gonzalez, S., Predominant effects of Polypodium leucotomos on membrane integrity, lipid peroxidation, and expression of elastin and matrixmetalloproteinase-1 in ultraviolet radiation exposed fibroblasts, and keratinocytes. Journal of Dermatological Science 2003, 32, (1), 1-9.
24.Gilchrest, B.; Yaar, M., Aging and photoaging of the skin: observations at the cellular and molecular level. The British Journal of Dermatology 1992, 127, (Suppl 41), 25-35.
25.Scharffetter-Kochanek, K.; Wlaschek, M.; Brenneisen, P.; Schauen, M.; Blaudschun, R.; Wenk, J., UV-induced reactive oxygen species in photocarcinogenesis and photoaging. Biological Chemistry 1997, 378, (11), 1247-1257.
26.Bernstein, E. F.; Uitto, J., The effect of photodamage on dermal extracellular matrix. Clinics in Dermatology 1996, 14, (2), 143-151.
27.JuttaWenk; Brenneisen, P.; Meewes, C.; Wlaschek, M.; Peters, T.; Blaudschun, R.; Ma, W.; Kuhr, L.; Schneider, L.; Scharffetter-Kochanek, K., UV-induced oxidative stress and photoaging. Current Problems in Dermatology 2001, 29, 83-94.
28.Yong, V. W.; Power, C.; Forsyth, P.; Edwards, D. R., Metalloproteinases in biology and pathology of the nervous system. Nature Reviews Neuroscience 2001, 2, 502-511.
29.Vu, T. H.; Werb, Z., Matrix metalloproteinases: effectors of development and normal physiology. Genes and Development 2000, 14, 2123-2133.
30.Rømer, J.; Bugge, T. H.; Fyke, C.; Lund, L. R.; Flick, M. J.; Degen, J. L.; Danø, K., Impaired wound healing in mice with a disrupted plasminogen gene. Nature Medicine 1996, 2, 287-292.
31.Page-McCaw, A.; Ewald, A. J.; Werb, Z., Matrix metalloproteinases and the regulation of tissue remodelling. Nature Reviews Molecular Cell Biology 2007, 8, 221-233.
32.Stetler-Stevenson, W. G., Matrix metalloproteinases in angiogenesis: a moving target for therapeutic intervention. The Journal of Clinical Investigation 1999, 103, (9), 1237-1241.
33.Nagase, H., Zinc Metalloproteases in Health and Disease(Hooper, N. M., ed). Taylor and Francis London 1996, 153-204.
34.Parks, W. C.; Mecham, R. P., Matrix metalloproteinases (Biology of Extracellular Matrix). Academic Press 1998, 1-362.
35.Martel-Pelletier, J.; McCollum, R.; Fujimoto, N.; Obata, K.; M.Cloutier, J.; Pelletier, J. P., Excess of metalloproteases over tissue inhibitor of metalloprotease may contribute to cartilage degradation in osteoarthritis and rheumatoid arthritis. Laboratory Investigation 1994, 70, (6), 807-815.
36.Dollery, C. M.; McEwan, J. R.; Henney, A. M., Matrix Metalloproteinases and Cardiovascular Disease Circulation Research 1995, 77, 863-868.
37.Sivak, J. M.; Fini, M. E., MMPs in the eye: emerging roles for matrix metalloproteinases in ocular physiology. Progress in Retinal and Eye Research 2002, 21, (1), 1-14.
38.Lenz, O.; Elliot, S. J.; Stetler-Stevenson, W. G., Matrix Metalloproteinases in Renal Development and Disease Journal of the American Society of Nephrology 2000, 11, 574-581.
39.Otani, Y.; Sakurai, Y.; Kameyama, K.; Igarashi, N.; Yokoyama, T.; Kubota, T.; Kumai, K.; Kitajima, M., Matrix Metalloproteinase Gene Expression in Chronic Gastric Ulcer: A Potential Role of Eosinophils in Perforation. Journal of Clinical Gastroenterology 1997, 25, S101-S104.
40.O'Connor, C. M.; FitzGerald, M. X., Matrix metalloproteases and lung disease. British Medical Journal 1994, 49, 602-609.
41.Arthur, M. J. P., Fibrogenesis II. Metalloproteinases and their inhibitors in liver fibrosis. American Journal of Physiology- Gastrointestinal and Liver Physiology 2000, 279, 245-249.
42.Ray, J. M.; Stetler-Stevenson, W. G., The role of matrix metalloproteases and their inhibitors in tumour invasion, metastasis and angiogenesis. European Respiratory Journal 1994, 7, 2062-2072.
43.Werb, C. C. a. Z., The many faces of metalloproteases: cell growth, invasion, angiogenesis and metastasis. Trends in Cell Biology 2001, 11, (11), S37-S43.
44.Wan, Y. S.; Wang, Z. Q.; Voorhees, J.; Fisher, G., EGF receptor crosstalks with cytokine receptors leading to the activation of c-Jun kinase in response to UV irradiation in human keratinocytes. Cellular Signalling 2001, 13, (2), 139-144.
45.Fisher, G. J.; Talwar, H. S.; Lin, J.; Voorhees, J. J., Molecular mechanisms of photoaging in human skin in vivo and their prevention by all-trans retinoic acid. Photochemistry and Photobiology 1999, 69, (2), 154-157.
46.Greul, A. K.; Grundmann, J. U.; Heinrich, F.; Pfitzner, I.; Bernhardt, J.; Ambach, A.; Biesalski, H. K.; Gollnick, H., Photoprotection of UV-irradiated human skin: an antioxidative combination of vitamins E and C, carotenoids, selenium and proanthocyanidins. Skin Pharmacology and Applied Skin Physiology 2002, 15, 307-315.
47.Fisher, G. J.; Datta, S. C.; Talwar, H. S.; Wang, Z. Q.; Varani, J.; Kang, S.; Voorhees, J. J., Molecular basis of sun-induced premature skin ageing and retinoid antagonism. Nature 1996, 379, 335-339.
48.Fisher, G. J.; Wang, Z.; Datta, S. C.; Varani, J.; Kang, S.; Voorhees, J. J., Pathophysiology of premature skin aging induced by ultraviolet light. The New England Journal of Medicine 1997, 337, (20), 1419-1428.
49.Griffith, L. G.; Swartz, M. A., Capturing complex 3D tissue physiology in vitro. Nature Reviews Molecular Cell Biology 2006, 7, 211-224.
50.Kähäri, V. M.; Kere, U. S., Matrix metalloproteinases in skin. Experimental Dermatology 1997, 6, (5), 199-213.
51.Pilcher, B. K.; Wang, M.; Qin, X. J.; Parks, W. C.; Senior, R. M.; Welgus, H. G., Role of matrix metalloproteinases and their inhibition in cutaneous wound healing and allergic contact hypersensitivity. Annals of the New York Academy of Sciences 1999, 878, 12-24.
52.Yaar, M.; Gilchrest, B. A., Photoageing: mechanism, prevention and therapy. British Journal of Dermatology 2007, 157, (5), 874-887.
53.Sternlicht, M. D.; ZenaWerb, How matrix metalloproteinases regulate cell behavior. Annual review of cell and developmental biology 2001, 17, 463-516.
54.Visse, R.; Nagase, H., Matrix Metalloproteinases and Tissue Inhibitors of Metalloproteinases: Structure, Function, and Biochemistry. Circulation Research 2003, 92, 827-839.
55.Overall, C. M.; López-Otín, C., Strategies for MMP inhibition in cancer: innovations for the post-trial era. Nature Reviews Cancer 2002, 2, 657-672.
56.Cury, P. R.; Araújo, V. C. d.; Canavez, F.; Furuse, C.; Leite, K. R. M.; Araújo, N. S. d., The effect of epidermal growth factor on matrix metalloproteinases and tissue inhibitors of metalloproteinase gene expression in cultured human gingival fibroblasts. Archives of Oral Biology 2007, 52, (6), 585-590.
57.Gross, J.; Lapiere, C. M., Collagenolytic activity in amphibian tissues: a tissue culture assay. Proceedings of the National Academy of Sciences 1962, 48, 1014-1022.
58.Greenlee, K. J.; Werb, Z.; Kheradmand, F., Matrix Metalloproteinases in Lung: Multiple, Multifarious, and Multifaceted. Physiological Reviews 2007, 87, 69-98.
59.Puente, X. S.; Pendás, A. M.; Llano, E.; Velasco, G.; López-Otín, C., Molecular cloning of a novel membrane-type matrix metalloproteinase from a human breast carcinoma. Cancer Research 1996, 56, (5), 944-949.
60.Lafleur, M. A.; Handsley, M. M.; Edwards, D. R., Metalloproteinases and their inhibitors in angiogenesis. Expert Reviews in Molecular Medicine 2003, 5, (23), 1-39.
61.Nagase, H.; Jr., J. F. W., Matrix metalloproteinases. Journal of Biological Chemistry 1999, 274, (31), 21491-21494.
62.Stolow, M. A.; Bauzon, D. D.; Li, J.; Sedgwick, T.; Liang, V. C.-T.; Sang, Q. A.; Shi, Y.-B., Identification and characterization of a novel collagenase in Xenopus laevis: possible roles during frog development. Molecular Biology of the Cell 1996, 7, (10), 1471-1483.
63.Hotary, K. B.; Allen, E. D.; Brooks, P. C.; Datta, N. S.; Long, M. W.; Weiss, S. J., Membrane type I matrix metalloproteinase usurps tumor growth control imposed by the three-dimensional extracellular matrix. Cell 2003, 114, (1), 33-45.
64.Somerville, R. P.; Oblander, S. A.; Apte, S. S., Matrix metalloproteinases: old dogs with new tricks. Genome Biology 2003, 4, (6), 216.
65.Parks, W. C.; Wilson, C. L.; López-Boado, Y. S., Matrix metalloproteinases as modulators of inflammation and innate immunity. Nature Reviews Immunology 2004, 4, (8), 617-629.
66.Welgus, H. G.; Stricklin, G. P.; Eisen, A. Z.; Bauer, E. A.; Cooney, R. V.; Jeffrey, J. J., A specific inhibitor of vertebrate collagenase produced by human skin fibroblasts. Journal of Biological Chemistry 1979, 254, (6), 1938-1943.
67.Docherty, A. J. P.; Lyons, A.; Smith, B. J.; Wright, E. M.; Stephens, P. E.; Harris, T. J. R.; Murphy, G.; Reynolds, J. J., Sequence of human tissue inhibitor of metalloproteinases and its identity to erythroidpotentiating activity. Nature 1985, 318, (7), 66-69.
68.Oh, J.; Takahashi, R.; Kondo, S.; Mizoguchi, A.; Adachi, E.; Sasahara, R. M.; Nishimura, S.; Imamura, Y.; Kitayama, H.; Alexander, D. B.; Ide, C.; Horan, T. P.; Arakawa, T.; Yoshida, H.; Nishikawa, S. i.; Itoh, Y.; Seiki, M.; Itohara, S.; Takahashi, C.; Noda, M., The membrane-anchored MMP inhibitor RECK is a key regulator of extracellular matrix integrity and angiogenesis. Cell 2001, 107, (6), 789-800.
69.Herman, M. P.; Sukhova, G. K.; Kisiel, W.; Foster, D.; Kehry, M. R.; Libby, P.; Schönbeck, U., Tissue factor pathway inhibitor-2 is a novel inhibitor of matrix metalloproteinases with implications for atherosclerosis. Journal of Clinical Investigation 2001, 107, (9), 1117-1126.
70.Mott, J. D.; Thomas, C. L.; Rosenbach, M. T.; Takahara, K.; Greenspan, D. S.; Banda, M. J., Post-translational proteolytic processing of procollagen C-terminal proteinase enhancer releases a metalloproteinase inhibitor. Journal of Biological Chemistry 2000, 275, (2), 1384-1390.
71.Pillai, S.; Oresajo, C.; Hayward, J., Ultraviolet radiation and skin aging: roles of reactive oxygen species, inflammation and protease activation, and strategies for prevention of inflammation-induced matrix degradation-a review. International Journal of Cosmetic Science 2005, 27, (1), 17-34.
72.Chirco, R.; Liu, X. W.; Jung, K. K.; Kim, H. R. C., Novel functions of TIMPs in cell signaling. Cancer and Metastasis Reviews 2006, 25, (1), 99-113.
73.Gomez, D. E.; Alonso, D. F.; Yoshiji, H.; Thorgeirsson, U. P., Tissue inhibitors of metalloproteinases: structure, regulation and biological functions. European journal of cell biology 1997, 74, (2), 111-122.
74.Sethi, C. S.; Bailey, T. A.; Luthert, P. J.; Chong, N. H. V., Matrix metalloproteinase biology applied to vitreoretinal disorders. British Journal of Ophthalmology 2000, 84, 654-666.
75.Murphy, G.; Willenbrock, F., Tissue inhibitors of matrix metalloendopeptidases. Methods in enzymology 1995, 248, 496-510.
76.R¨th, F. X. G.; Maskos, K.; Betz, M.; Bergner, A.; Huber, R.; Suzuki, K.; Yoshida, N.; Nagase, H.; Brew, K.; Bourenkov, G. P.; Bartunik, H.; Bode, W., Mechanism of inhibition of the human matrix metalloproteinase stromelysin-1 by TIMP-1. Nature 1997, 389, (4), 77-81.
77.Webster, G. L., Classification of the Euphorbiaceae. Annals of the Missouri Botanical Garden 1994, 81, (1), 3-32.
78.李時珍, 本草綱目. 漢珍數位圖書公司 明代.
79.吳征鎰, 新華本草綱要. 上海科學技術出版社 1991, 第二冊.
80.李同琴; 郭秋紅; 田質芬, 大戟科植物藥用歷史沿革及價值的探討. 中醫藥學刊 2003, 21, (8), 1349-1350.
81.行政院衛生署中醫藥委員會, 臺灣藥用植物資源名錄. 2003, 285.
82.黃增泉; 耿煊; 謝萬權; 蔡進來; 胡哲明; 沈中桴; 楊國禎; 謝長富等, 台灣植物誌. Flora of Taiwan 1993, 3, 461.
83.Cos, P.; Ying, L.; Calomme, M.; Hu, J. P.; Cimanga, K.; Poel, B. V.; Pieters, L.; Vlietinck, A. J.; Berghe, D. V., Structure-Activity Relationship and Classification of Flavonoids as Inhibitors of Xanthine Oxidase and Superoxide Scavengers. Journal of Natural Products 1998, 61, (1), 71-76.
84.Crespo, I.; García-Mediavilla, M. V.; Almar, M.; González, P.; Tuñón, M. J.; Sánchez-Campos, S.; González-Gallego, J., Differential effects of dietary flavonoids on reactive oxygen and nitrogen species generation and changes in antioxidant enzyme expression induced by proinflammatory cytokines in Chang Liver cells. Food and Chemical Toxicology 2008, 46, (5), 1555-1569.
85.Kampkötter, A.; Nkwonkam, C. G.; Zurawski, R. F.; Timpel, C.; Chovolou, Y.; Wätjen, W.; Kahl, R., Effects of the flavonoids kaempferol and fisetin on thermotolerance, oxidative stress and FoxO transcription factor DAF-16 in the model organism Caenorhabditis elegans. Archives of Toxicology 2007, 81, (12), 849-858.
86.Takasawa, R.; Takahashi, S.; Saeki, K.; Sunaga, S.; Yoshimori, A.; Tanuma, S. i., Structure-activity relationship of human GLO I inhibitory natural flavonoids and their growth inhibitory effects Bioorganic & Medicinal Chemistry 2007, 16, (7), 3969-3975.
87.Lin, R. D.; Chin, Y. P.; Hou, W. C.; Lee, M. H., The Effects of Antibiotics Combined with Natural Polyphenols against Clinical Methicillin-Resistant Staphylococcus aureus (MRSA). Planta Medica 2008, 74, (8), 840-846.
88.Liu, A. L.; Wang, H. D.; Lee, S. M.; Wang, Y. T.; Du, G. H., Structure-activity relationship of flavonoids as influenza virus neuraminidase inhibitors and their in vitro anti-viral activities. Bioorganic and Medicinal Chemistry 2008, 16, (15), 7141-7147.
89.Serkedjieva, J.; Gegova, G.; Mladenov, K., Protective efficacy of an aerosol preparation, obtained from Geranium sanguineum L., in experimental influenza infection. Pharmazie 2008, 63, (2), 160-163.
90.Leung, H. W. C.; Lin, C. J.; Hour, M. J.; Yang, W. H.; Wang, M. Y.; Lee, H. Z., Kaempferol induces apoptosis in human lung non-small carcinoma cells accompanied by an induction of antioxidant enzymes Food and Chemical Toxicology 2007, 45, (10), 2005-2013.
91.Park, E. J.; Kim, Y.; Kim, J., Acylated Flavonol Glycosides from the Flower of Inula britannica. Journal of Natural Products 2000, 63, (1), 34-36.
92.Hou, L.; Zhou, B.; Yang, L.; Liu, Z. L., Inhibition of free radical initiated peroxidation of human erythrocyte ghosts by flavonols and their glycosides. Organic and Biomolecular Chemistry 2004, 2, (9), 1419-1423.
93.Ibrahim, L. F.; El-Senousy, W. M.; Hawas, U. W., NMR spectral analysis of flavonoids from Chrysanthemum coronarium Chemistry of Natural Compounds 2007, 43, (6), 659-662.
94.Wei, F.; Ma, S. C.; Ma, L. Y.; But, P. P. H.; Lin, R. C.; Khan, I. A., Antiviral Flavonoids from the Seeds of Aesculus chinensis. Journal of Natural Products 2004, 67, (4), 650-653.
95.Wang, A. X.; Zhang, Q.; Jia, Z. J., A new furobenzopyranone and other constituents from Anaphalis lactea. Pharmazie 2004, 59, (10), 807-811.
96.Hsu, H. Y.; Tsang, S. F.; Lin, K. W.; Yang, S. C.; Lin, C. N., Cell death induced by flavonoid glycosides with and without copper. Food and Chemical Toxicology 2008, 46, (7), 2394-2401.
97.Zhu, J. T. T.; Choi, R. C. Y.; Chu, G. K. Y.; Cheung, A. W. H.; Gao, Q. T.; Li, J.; Jiang, Z. Y.; Dong, T. T. X.; Tsim, K. W. K., Flavonoids Possess Neuroprotective Effects on Cultured Pheochromocytoma PC12 Cells: A Comparison of Different Flavonoids in Activating Estrogenic Effect and in Preventing β-Amyloid-Induced Cell Death. Journal of Agricultural and Food Chemistry 2007, 55, (6), 2438-2445.
98.Wang, L.; Tu, Y. C.; Lian, T. W.; Hung, J. T.; Yen, J. H.; Wu, M. J., Distinctive Antioxidant and Antiinflammatory Effects of Flavonols. Journal of Agricultural and Food Chemistry 2006, 54, (26), 9798-9804.
99.Belinha, I.; Amorim, M. A.; Rodrigues, P.; Freitas, V. d.; Moradas-Ferreira, P.; Mateus, N.; Costa, V., Quercetin Increases Oxidative Stress Resistance and Longevity in Saccharomyces cerevisiae. Journal of Agricultural and Food Chemistry 2007, 55, (6), 2446-2451.
100.deGraft-Johnson, J.; Kolodziejczyk, K.; Krol, M.; Nowak, P.; Krol, B.; Nowak, D., Ferric-Reducing Ability Power of Selected Plant Polyphenols and Their Metabolites: Implications for Clinical Studies on the Antioxidant Effects of Fruits and Vegetable Consumption. Basic and Clinical Pharmacology and Toxicology 2007, 100, (5), 345-352.
101.Mamani-Matsuda, M.; Kauss, T.; Al-Kharrat, A.; Rambert, J.; Fawaz, F.; Thiolat, D.; Moynet, D.; Coves, S.; Malvy, D.; Mossalayi, M. D., Therapeutic and preventive properties of quercetin in experimental arthritis correlate with decreased macrophage inflammatory mediators Biochemical Pharmacology 2006, 72, (10), 1304-1310.
102.Indap, M.; Radhika, S.; Motiwale, L.; Rao2, K., Quercetin: Antitumor activity and pharmacological manipulations for increased therapeutic gains. Indian Journal of Pharmaceutical Sciences 2006, 68, (4), 465-469.
103.TM, E.; AS, V., The inhibitory effect of curcumin, genistein, quercetin and cisplatin on the growth of oral cancer cells in vitro. Anticancer Research 2000, 20, (3A), 1733-1738.
104.Bravo, A.; Anacona, J. R., Metal complexes of the flavonoid quercetin: antibacterial properties Transition Metal Chemistry 2001, 26, (1-2), 20-23.
105.Aziz, N.; Farag, S.; Mousa, L.; Abo-Zaid, M., Comparative antibacterial and antifungal effects of some phenolic compounds Microbios 1998, 93, (374), 34-54.
106.Lin, R.; Liu, J.; Gan, W.; Ding, C., Protective Effect of Quercetin on the Homocysteine-Injured Human Umbilical Vein Vascular Endothelial Cell Line (ECV304). Basic and Clinical Pharmacology and Toxicology 2007, 101, (3), 197-202.
107.Okamura, H.; Mimura, A.; Yakou, Y.; Niwano, M.; Takahara, Y., Antioxidant activity of tannins and flavonoids in Eucalyptus rostrata Phytochemistry 1993, 33, (3), 557-561.
108.Haraguchi, H.; Ishikawa, H.; Sanchez, Y.; Ogura, T.; Kubo, Y.; Kubo, I., Antioxidative constituents in Heterotheca inuloides Bioorganic and Medicinal Chemistry 1997, 5, (5), 865-871.
109.Cimanga, K.; Ying, L.; Bruyne, T. d.; Apers, S.; Cos, P.; Hermans, N.; Bakana, P.; Tona, L.; Kambu, K.; Kalenda, D. T.; Pieters, L.; Berghe, D. v.; Vlietinck, A. J., Radical scavenging and xanthine oxidase inhibitory activity of phenolic compounds from Bridelia ferruginea stem bark Journal of Pharmacy and Pharmacology 2001, 53, (5), 747-761.
110.Wu, Q. L.; Wang, S. P.; Du, L. J.; Zhang, S. M.; Yang, J. S.; Xiao, P. G., Chromone glycosides and flavonoids from hypericum japonicum Phytochemistry 1998, 49, (5), 1417-1420.
111.Park, K. Y.; Lee, S. H.; Min, B. K.; Lee, K. S.; Choi, J. S.; Chung, S. R.; Min, K. R.; Kim, Y. s., Inhibitory Effect of Luteolin 4'-O-Glucoside from Kummerowia striata and Other Flavonoids on Interleukin-5 Bioactivity. Planta Medica 1999, 65, (5), 457-459.
112.Ramiro, E.; Franch, À.; Castellote, C.; Pérez-Cano, F.; Permanyer, J.; Izquierdo-Pulido, M.; Castell, M., Flavonoids from Theobroma cacao Down-Regulate Inflammatory Mediators. Journal of Agricultural and Food Chemistry 2005, 53, (22), 8506-8511.
113.Murakami, N.; Mostaqul, H. M.; Tamura, S.; Itagaki, S.; Horii, T.; Kobayashi, M., New anti-malarial flavonol glycoside from Hydrangeae dulcis folium Bioorganic and Medicinal Chemistry letters 2001, 11, (18), 2445-2447.
114.Mastuda, H.; Morikawa, T.; Ueda, K.; Managi, H.; Yoshikawa, M., Structural requirements of flavonoids for inhibition of antigen-Induced degranulation, TNF-α and IL-4 production from RBL-2H3 cells Bioorganic and Medicinal Chemistry 2002, 10, (10), 3123-3128.
115.He, X.; Liu, R. H., Cranberry Phytochemicals: Isolation, Structure Elucidation, and Their Antiproliferative and Antioxidant Activities. Journal of Agricultural and Food Chemistry 2006, 54, (19), 7069-7074.
116. Snijman, P. W.; Swanevelder, S.; Joubert, E.; Green, I. R.; Gelderblom, W. C. A., The antimutagenic activity of the major flavonoids of rooibos (Aspalathus linearis): Some dose-response effects on mutagen activation-flavonoid interactions. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 2007, 631, (2), 111-123.
117.Ito, M.; Shimura, H.; Watanabe, N.; Tamai, M.; Hanada, K., Hepatoprotective Compounds from Canarium album and Euphorbia nematocypha. Chemical & Pharmaceutical Bulletin 1990, 38, (8), 2201-2203.
118.Quettier-Deleu, C.; Voiselle, G.; Fruchart, J. C.; Duriez, P.; Teissier, E.; Bailleul, F.; Vasseur, J.; Trotin, F., Hawthorn extracts inhibit LDL oxidation. Pharmazie 2003, 58, (8), 577-581.
119.Butterweck, V.; Jürgenliemk, G.; Nahrstedt, A.; Winterhoff, H., Flavonoids from Hypericum perforatum Show Antidepressant Activity in the Forced Swimming Test. Planta Medica 2000, 66, (1), 3-6.
120.Xiong, Q.; Fan, W.; Tezuka, Y.; Adnyana, I. K.; Stampoulis, P.; Hattori, M.; Namba, T.; Kadota, S., Hepatoprotective Effect of Apocynum venetum and its Active Constituents. Planta Medica 2000, 66, (2), 127-133.
121.Hammer, K. D. P.; Hillwig, M. L.; Solco, A. K. S.; Dixon, P. M.; Delate, K.; Murphy, P. A.; Wurtele, E. S.; Birt, D. F., Inhibition of Prostaglandin E2 Production by Anti-inflammatory Hypericum perforatum Extracts and Constituents in RAW264.7 Mouse Macrophage Cells. Journal of Agricultural and Food Chemistry 2007, 55, (18), 7323-7331.
122.Hernandez, J.; Goycoolea, F. M.; Quintero, J.; Acosta, A.; Castañeda, M.; Dominguez, Z.; Robles, R.; Vazquez-Moreno, L.; Velazquez, E. F.; Astiazaran, H.; Lugo, E.; Velazquez, C., Sonoran Propolis: Chemical Composition and Antiproliferative Activity on Cancer Cell Lines. Planta Medica 2007, 73, (14), 1469-1474.
123.Khushbaktova, Z. A.; Faizieva, S. K.; Syrov, V. N.; Yuldashev, M. P.; Batirov, É. K.; Mamatkhanov, A. U., Isolation, Chemical Analysis, and Study of the Hypolipidemic Activity of the Total Flavonoid Extract from Thermopsis altherniaflora Pharmaceutical Chemistry Journal 2001, 35, (3), 155-158.
124.Fukai, T.; Marumo, A.; Kaitou, K.; Kanda, T.; Terada, S.; Nomura, T., Antimicrobial activity of licorice flavonoids against methicillin-resistant Staphylococcus aureus Fitoterapia 2002, 73, (6), 536-539.
125.Navarro, A.; Heras, B. D.; Villar, A., Anti-Inflammatory and Immunomodulating Properties of a Sterol Fraction from Sideritis foetens Clem. Biological and Pharmaceutical Bulletin 2001, 24, (5), 470-473.
126.Jin, J. L.; Lee, S.; Lee, Y. Y.; Kim, J. M.; Heo, J. E.; Yun-Choi, H. S., Platelet Anti-Aggregating Triterpenoids from the Leaves of Acanthopanax senticosus and the Fruits of A. sessiliflorus. Planta Medica 2004, 70, (6), 564-566.
127.Ali, M. S.; Banskota, A. H.; Tezuka, Y.; Saiki, I.; Kadota, S., Antiproliferative Activity of Diarylheptanoids from the Seeds of Alpinia blepharocalyx. Biological and Pharmaceutical Bulletin 2001, 24, (5), 525-528.
128.Ali, Z.; Khan, S. I.; Khan, I. A., Phytochemical Study of Actaea rubra and Biological Screenings of Isolates. Planta Medica 2006, 72, (14), 1350-1352.
129.Akihisa, T.; Yasukawa, K.; Oinuma, H.; Kasahara, Y.; Yamanouchi, S.; Takido, M.; Kumaki, K.; Tamura, T., Triterpene alcohols from the flowers of compositae and their anti-inflammatory effects. Phytochemistry 1996, 43, (6), 1255-1260.
130.Saeed, M. A.; Sabir, A. W., Antibacterial activity of Caesalpinia bonducella seeds. Fitoterapia 2001, 72, (7), 807-809.
131.Jain, S. C.; Singh, B.; Jain, R., Antimicrobial activity of triterpenoids from Heliotropium ellipticum Fitoterapia 2001, 72, (6), 666-668.
132.Akihisa, T.; Franzblau, S. G.; Ukiya, M.; Okuda, H.; Zhang, F.; Yasukawa, K.; Suzuki, T.; Kimura, Y., Antitubercular Activity of Triterpenoids from Asteraceae Flowers. Biological & Pharmaceutical Bulletin 2005, 28, (1), 158-160.
133.Bringmann, G.; Hamm, A.; Günther, C.; Michel, M.; Brun, R.; Mudogo, V., Ancistroealaines A and B, Two New Bioactive Naphthylisoquinolines, and Related Naphthoic Acids from Ancistrocladus ealaensis. Journal of Natural Products 2000, 63, (11), 1465-1470.
134.Dong, M.; He, X.; Liu, R. H., Phytochemicals of Black Bean Seed Coats: Isolation, Structure Elucidation, and Their Antiproliferative and Antioxidative Activities. Journal of Agricultural and Food Chemistry 2007, 55, (15), 6044-6051.
135.Zou, J. H.; Dai, J.; Chen, X.; Yuan, J. Q., Pentacyclic Triterpenoids from Leaves of Excoecaria agallocha. Chemical & Pharmaceutical Bulletin 2006, 54, (6), 920-921.
136.Kuete, V.; Eyong, K. O.; Folefoc, G. N.; Beng, V. P.; Hussain, H.; Krohn, K.; Nkengfack, A. E., Antimicrobial activity of the methanolic extract and of the chemical constituents isolated from Newbouldia laevis. Pharmazie 2007, 62, (7), 552-556.
137.Horiuchi, K.; Shiota, S.; Hatano, T.; Yoshida, T.; Kuroda, T.; Tsuchiya, T., Antimicrobial Activity of Oleanolic Acid from Salvia officinalis and Related Compounds on Vancomycin-Resistant Enterococci (VRE). Biological and Pharmaceutical Bulletin 2007, 30, (6), 1147-1149.
138.Cipak, L.; Grausova, L.; Miadokova, E.; Novotny, L.; Rauko, P., Dual activity of triterpenoids: apoptotic versus antidifferentiation effects Archives of Toxicology 2006, 80, (7), 429-435.
139.Fu, L.; Zhang, S.; Li, N.; Wang, J.; Zhao, M.; Sakai, J.; Hasegawa, T.; Mitsui, T.; Kataoka, T.; Oka, S.; Kiuchi, M.; Hirose, K.; Ando, M., Three New Triterpenes from Nerium oleander and Biological Activity of the Isolated Compounds. Journal of Natural Products 2005, 68, (2), 198-206.
140.Son, J. K.; Jung, S. J.; Jung, J. H.; Fang, Z.; Lee, C. S.; Seo, C. S.; Moon, D. C.; Min, B. S.; Kim, M. R.; Woo, M. H., Anticancer Constituents from the Roots of Rubia cordifolia L. Chemical and Pharmaceutical Bulletin 2008, 56, (2), 213-216.
141.Rodriguez-Rodriguez, R.; Stankevicius, E.; Herrera, M. D.; Østergaard, L.; Andersen, M. R.; Ruiz-Gutierrez, V.; Simonsen, U., Oleanolic acid induces relaxation and calcium-independent release of endothelium-derived nitric oxide. British Journal of Pharmacology 2008, 155, (4), 535-546.
142.Takasaki, M.; Konoshima, T.; Tokuda, H.; Masuda, K.; Arai, Y.; Shiojima, K.; Ageta, H., Anti-carcinogenic Activity of Taraxacum Plant. II. Biological & Pharmaceutical Bulletin 1999, 22, (6), 606-610.
143.Chaturvedula, V. S. P.; Schilling, J. K.; Malone, S.; Wisse, J. H.; Werkhoven, M. C. M.; Kingston, D. G. I., New Cytotoxic Triterpene Acids from Aboveground Parts of Manihot esculenta from the Suriname Rainforest. Planta Medica 2003, 69, (3), 271-274.
144.Kanlayavattanakul, M.; Ruangrungsi, N.; Watanabe, T.; Kawahata, M.; Therrien, B.; Yamaguchi, K.; Ishikawa, T., ent-Halimane Diterpenes and a Guaiane Sesquiterpene from Cladogynos orientalis. Journal of Natural Products 2005, 68, (1), 7-10.
145.Naik, D. G.; Mujumdar, A. M.; Waghole, R. J.; Misar, A. V.; Bligh, S. W. A.; Bashall, A.; Crowder, J., Taraxer-14-en-3β-ol, an Anti-Inflammatory Compound from Sterculia foetida L. Planta Medica 2004, 70, (1), 68-69.
146.Duwiejua, M.; Zeitlin, I. J.; Gray, A. I.; Waterman, P. G., The Anti-Inflammatory Compounds of Polygonum bistorta: Isolation and Characterisation. Planta Medica 1999, 65, (4), 371-374.
147.Choudhary, M. I.; Azizuddin; Jalil, S.; Atta-ur-Rahman, Bioactive phenolic compounds from a medicinal lichen, Usnea longissima Phytochemistry 2005, 66, (19), 2346-2350.
148.Moiteiro, C.; Justino, F.; Tavares, R.; Marcelo-Curto, M. J.; Florêncio, M. H.; Nascimento, M. S. J.; Pedro, M.; Cerqueira, F.; Pinto, M. M. M., Synthetic Secofriedelane and Friedelane Derivatives as Inhibitors of Human Lymphocyte Proliferation and Growth of Human Cancer Cell Lines in Vitro. Journal of Natural Products 2001, 64, (10), 1273-1277.
149.Reyes-Chilp, R.; Estrada-Muñiz, E.; Apan, T. R.; Amekraz, B.; Aumelas, A.; Jankowski, C. K.; Vázquez-Torres, M., Cytotoxic effects of mammea type coumarins from Calophyllum brasiliense. Life Sciences 2004, 75, (13), 1635-1647.
150.Chang, K. C.; Duh, C. Y.; Chen, I. S.; Tsai, I. L., A Cytotoxic Butenolide, Two New Dolabellane Diterpenoids, a Chroman and a Benzoquinol Derivative Formosan Casearia membranacea. Planta Medica 2003, 69, (7), 667-672.
151.Mitaine-Offer, A. C.; Sauvain, M.; Deharo, E.; Muñoz, V.; Zèches-Hanrot, M., A New Diterpene from Tanaecium jaroba. Planta Medica 2002, 68, 568-569.
152.Lenta, B. N.; Devkota, K. P.; Ngouela, S.; Boyom, F. F.; Naz, Q.; Choudhary, M. I.; Tsamo, E.; Rosenthal, P. J.; Sewald, N., Anti-plasmodial and Cholinesterase Inhibiting Activities of some Constituents of Psorospermum glaberrimum. Chemical and Pharmaceutical Bulletin 2008, 56, (2), 222-226.
153.Fukuda, T.; Ito, H.; Yoshida, T., Antioxidative polyphenols from walnuts (Juglans regia L.) Phytochemistry 2003, 63, (7), 795-801.
154.Dong, H.; Chen, S. X.; Kini, R. M.; Xu, H. X., Effects of Tannins from Geum japonicum on the Catalytic Activity of Thrombin and Factor Xa of Blood Coagulation Cascade. Journal of Natural Products 1998, 61, (11), 1356-1360.
155.Mook-Jung, I.; Kim, H.; Fan, W.; Tezuka, Y.; Kadota, S.; Nishijo, H.; Jung, M. W., Neuroprotective Effects of Constituents of the Oriental Crude Drugs, Rhodiola sacra, R. sachalinensis and Tokaku-joki-to, against Beta-amyloid Toxicity, Oxidative Stress and Apoptosis. Biological & Pharmaceutical Bulletin 2002, 25, (8), 1101-1104.
156.Kurihara, H.; Shibata, H.; Fukui, Y.; Kiso, Y.; Xu, J. K.; Yao, X. S.; Fukami, H., Evaluation of the Hypolipemic Property of Camellia sinensis Var. ptilophylla on Postprandial Hypertriglyceridemia. Journal of Agricultural and Food Chemistry 2006, 54, (14), 4977-4981.
157.Chen, W. J.; Chang, C. Y.; Lin, J. K., Induction of G1 phase arrest in MCF human breast cancer cells by pentagalloylglucose through the down-regulation of CDK4 and CDK2 activities and up-regulation of the CDK inhibitors p27Kip and p21Cip Biochemical Pharmacology 2003, 65, (11), 1777-1785.
158.Ho, L. L.; Chen, W. J.; Lin-Shiau, S. Y.; Lin, J. K., Penta-O-galloyl-β-d-glucose inhibits the invasion of mouse melanoma by suppressing metalloproteinase-9 through down-regulation of activator protein-1 European Journal of Pharmacology 2002, 453, (2-3), 149-158.
159.Lee, S. J.; Lee, H. K.; Jung, M. K.; Mar, W., In Vitro Antiviral Activity of 1,2,3,4,6-Penta-O-galloyl-β-D-glucose against Hepatitis B Virus. Biological & Pharmacetical Bulletin 2006, 29, (10), 2131-2134.
160.Lee, S. H.; Park, H. H.; Kim, J. E.; Kim, J. A.; Kim, Y. H.; Jun, C. D.; Kim, S. H., Allose Gallates Suppress Expression of Pro-Inflammatory Cytokines through Attenuation of NF-κB in Human Mast Cells. Planta Medica 2007, 73, (8), 769-773.
161.Valencia, E.; Valenzuela, E.; Barros, E.; Aedo, V.; Gebauer, M. T.; García, C.; González, A. G.; Bermejo, J., Constituents of Coriaria ruscifolia fruits Fitoterapia 2001, 72, (5), 555-557.
162.Atta-Ur-Rahman; Ngounou, F. N.; Choudhary, M. I.; Malik, S.; Makhmoor, T.; Nur-E-Alam, M.; Zareen, S.; Lontsi, D.; Ayafor, J. F.; Sondengam, B. L., New Antioxidant and Antimicrobial Ellagic Acid Derivatives from Pteleopsis hylodendron. Planta Medica 2001, 67, (4), 335-339.
163.Wang, R. F.; Xie, W. D.; Zhang, Z.; Xing, D. M.; Ding, Y.; Wang, W.; Ma, C.; Du, L. J., Bioactive Compounds from the Seeds of Punica granatum (Pomegranate). Journal of Natural Products 2004, 67, (12), 2096-2098.
164.Sinigaglia, M.; Lehmann, M.; Baumgardt, P.; Amaral, V. S. d.; Dihl, R. R.; Reguly, M. L.; Andrade, H. H. R. d., Vanillin as a modulator agent in SMART test: Inhibition in the steps that precede N-methyl-N-nitrosourea-, N-ethyl-N-nitrosourea-, ethylmethanesulphonate- and bleomycin-genotoxicity Mutation Research/Genetic Toxicology and Environmental Mutagenesis 2006, 607, (2), 225-230.
165.Ueda, J. y.; Awale, S.; Tezuka, Y.; Shimamura, E.; Hirai, K. i.; Nobukawa, T.; Sato, A.; Kadota, S., Growth Inhibitory Activity of Wood of Taxus yunnanensis and its Liquid Chromatography Fourier-Transform Mass Spectrometry Analysis. Planta Medica 2006, 72, (13), 1241-1244.
166.Lee, C. L.; Huang, P. C.; Hsieh, P. W.; Hwang, T. L.; Hou, Y. Y.; Chang, F. R.; Wu, Y. C., (-)-Xanthienopyran, a New Inhibitor of Superoxide Anion Generation by Activated Neutrophils, and Further Constituents of the Seeds of Xanthium strumarium. Planta Medica 2008, 74, (10), 1276-1279.
167.Lirdprapamongkol, K.; Sakurai, H.; Kawasaki, N.; Choo, M.-K.; Saitoh, Y.; Aozuka, Y.; Singhirunnusorn, P.; Ruchirawat, S.; Svasti, J.; Saiki, I., Vanillin suppresses in vitro invasion and in vivo metastasis of mouse breast cancer cells. European Journal of Pharmaceutical Sciences 2005, 25, (1), 57-65.
168.Chen, J. J.; Chou, E. T.; Duh, C. Y.; Yang, S. Z.; Chen, I. S., New Cytotoxic Tetrahydrofuran- and Dihydrofuran-Type Lignans from the Stem of Beilschmiedia tsangii. Planta Medica 2006, 72, (4), 351-357.
169.Harish, R.; Divakar, S.; Srivastava, A.; Shivanandappa, T., Isolation of Antioxidant Compounds from the Methanolic Extract of the Roots of Decalepis hamiltonii (Wight and Arn.). Journal of Agricultural and Food Chemistry 2005, 53, (20), 7709-7714.
170.Kanegae, M. P. P.; Fonseca, L. M. d.; Brunetti, I. L.; Silva, S. d. O.; Ximenes, V. F., The reactivity of ortho-methoxy-substituted catechol radicals with sulfhydryl groups: Contribution for the comprehension of the mechanism of inhibition of NADPH oxidase by apocynin. Biochemical Pharmacology 2007, 74, (3), 457-464.
171.Epifano, F.; Genovese, S.; Sosa, S.; Tubaro, A.; Curini, M., Synthesis and anti-inflammatory activity of 3-(4'-geranyloxy-3'-methoxyphenyl)-2-trans propenoic acid and its ester derivatives. Bioorganic and Medicinal Chemistry Letters 2007, 17, (20), 5709-5714.
172.Fang, J. J.; Ye, G.; Chen, W. L.; Zhao, W. M., Antibacterial phenolic components from Eriocaulon buergerianum Phytochemistry 2008, 69, (5), 1279-1286.
173.Tian, X. Y.; Wang, Y. H.; Liu, H. Y.; Yu, S. S.; Fang, W. S., On the Chemical Constituents of Dipsacus asper. Chemical and Pharmaceutical Bulletin 2007, 55, (12), 1677-1681.
174.Yang, C.; Shi, Y. P.; Jia, Z. J., Sesquiterpene Lactone Glycosides, Eudesmanolides, and Other Constituents from Carpesium macrocephalum. Planta Medica 2002, 68, 626-630.
175.Wu, T. S.; Shi, L. S.; Wang, J. J.; Iou, S. C.; Chang, H. C.; Chen, Y. P.; Kuo, Y. H.; Chang, Y. L.; Teng, C. M., Cytotoxic and Antiplatelet Aggregation Principles of Ruta graveolens. Journal of the Chinese Chemical Society 2003, 50, (1), 171-178.
176.Gu, J. Q.; Graf, T. N.; Lee, D.; Chai, H. B.; Mi, Q.; Kardono, L. B. S.; Setyowati, F. M.; Ismail, R.; Riswan, S.; Farnsworth, N. R.; Cordell, G. A.; Pezzuto, J. M.; Swanson, S. M.; Kroll, D. J.; Falkinham, J. O.; Wall, M. E.; Wani, M. C.; Kinghorn, A. D.; Oberlies, N. H., Cytotoxic and Antimicrobial Constituents of the Bark of Diospyros maritima Collected in Two Geographical Locations in Indonesia. Journal of Natural Products 2004, 67, (7), 1156-1161.
177.Dall'Acqua, S.; Viola, G.; Piacente, S.; Cappelletti, E. M.; Innocenti, G., Cytotoxic Constituents of Roots of Chaerophyllum hirsutum. Journal of Natural Products 2004, 67, (9), 1588-1590.
178.Abas, F.; Lajis, N. H.; Shaari, K.; Israf, D. A.; Stanslas, J.; Yusuf, U. K.; Raof, S. M., A Labdane Diterpene Glucoside from the Rhizomes of Curcuma mangga. Journal of Natural Products 2005, 68, (7), 1090-1093.
179.Kim, E. K.; Kwon, K. B.; Shin, B. C.; Seo, E. A.; Lee, Y. R.; Kim, J. S.; Park, J. W.; Park, B. H.; Ryu, D. G., Scopoletin induces apoptosis in human promyeloleukemic cells, accompanied by activations of nuclear factor κB and caspase-3 Life Sciences 2005, 77, (7), 824-836.
180.Chen, J. J.; Wang, T. Y.; Hwang, T. L., Neolignans, a Coumarinolignan, Lignan Derivatives, and a Chromene: Anti-inflammatory Constituents from Zanthoxylum avicennae. Journal of Natural Products 2008, 71, (2), 212-217.
181.Felczak, K.; Drabikowska, A. K.; Vilpo, J. A.; Kulikowski, T.; Shugar, D., 6-Substituted and 5,6-Disubstituted Derivatives of Uridine: Stereoselective Synthesis, Interaction with Uridine Phosphorylase, and in Vitro Antitumor Activity. Journal of Medicinal Chemistry 1996, 39, (8), 1720-1728.
182.Lee, C. L.; Chang, F. R.; Hsieh, P. W.; Chiang, M. Y.; Wu, C. C.; Huang, Z. Y.; Lan, Y. H.; Chen, M.; Lee, K. H.; Yen, H. F.; Hung, W. C.; Wu, Y. C., Cytotoxic ent-abietane diterpenes from Gelonium aequoreum. Phytochemistry 2008, 69, (1), 276-287.
183.Shizuri, Y.; Kosemura, S.; Yamamura, S.; Ohba, S.; Ito, M.; Saito, Y., Isolation and Structures of Helioscopinolide, New Diterpenes from Euphorbia helioscopia L. Chemistry Letters 1983, 12, (1), 65-68.
184.Crespi-Perellino, N.; Garofano, L.; Arlandini, E.; Pinciroli, V.; Minghetti, A.; Vincieri, F. F.; Danieli, B., Identification of New Diterpenoids from Euphorbia calyptrata Cell Cultures. Journal of Natural Products 1996, 59, (8), 773-776.
185.Borghi, D.; Baumer, L.; Ballabio, M.; Arlandini, E.; Perellino, N. C.; Minghetti, A.; Vincieri, F. F., Structure Elucidation of Helioscopinolides D and E from Euphorbia calyptrata Cell Cultures. Journal of Natural Products 1991, 54, (6), 1503-1508.
186.Yan, R. Y.; Tan, Y. X.; Cui, X. Q.; Chen, R. Y.; Yu, D. Q., Diterpenoids from the Roots of Suregada glomerulata. Journal of Natural Products 2008, 71, (2), 195-198.
187.He, F.; Pu, J. X.; Huang, S. X.; Xiao, W. L.; Yang, L. B.; Li, X. N.; Zhao, Y.; Ding, J.; Xu, C. H.; Sun, H. D., Eight New Diterpenoids from the Roots of Euphorbia nematocypha. Helvetica Chimica Acta 2008, 91, (11), 2139-2147.
188.Jamal, A. K.; Yaacob, W. A.; Din, L. B., A Chemical Study on Phyllanthus Columnaris. European Journal of Scientific Research 2009, 28, (1), 76-81.
189.Ageta, H.; Arai, Y.; Suzuki, H.; Kiyotani, T.; Kitabayashi, M., NMR Spectra of Triterpenoids. III. Oleanenes and Migrated Oleanenes. Chemical and Pharmaceutical Bulletin 1995, 43, (2), 198-203.
190.Olea, R. S. G.; Torres, L. M. B.; Roque, L. C.; Roque, N. F., I3C NMR Spectroscopic Data for Glutinol and Derivatives. Magnetic Resonance in Chemistry 1994, 32, (6), 378-379.
191.Antoniog.Gonzalez; Ferro, E.; Ravelo, A., Triterpenes from Maytenus horrida. Phytochemistry 1987, 26, (10), 2785-2788.
192.Matsunaga, S.; Tanaka, R.; Akagi, M., Triterpenoids from Euphorbia maculata. Phytochemistry 1988, 27, (2), 535-537.
193.Leong, Y. W.; Harrison, L. J., (20R,23E)-Eupha-8,23-diene-3β,25-diol from Tripetalum cymosum. Phytochemistry 1999, 50, (5), 849-857.
194.Lin, J. H.; Ku, Y. R.; Lin, Y. T.; Teng, S. F.; Wen, K. C.; Liao, C. H., Preparative Isolation and Gas Chromatography-Mass Spectrometry Analysis of Triterpenoids in Kansui Radix. Journal of Food and Drug Analysis 2000, 8, (4), 278-282.
195.Lavie, D.; Shvo, Y.; Glower, E., TERPENOIDS II - The Nuclear Magnetic Resonance Spectra of Tetracyclic Triterpenes. Tetrahedron 1963, 19, (12), 2255-2259.
196.Lee, S. C.; Kwon, Y. S.; Son, K. H.; Kim, H. P.; Heo, M. Y., Antioxidative Constituents from Paeonia lactiflora. Archives of Pharmacal Research 2005, 28, (7), 775-783.
197.Vasconcelos, J. M. J.; Silva, A. M. S.; Cavaleiro, J. A. S., Chromones and flavanones from artemisia campestris subsp. maritima. Phytochemistry 1998, 49, (5), 1421-1424.
198.Duarte, N. l.; Ferreira, M. J. U., Lagaspholones A and B: Two New Jatropholane-Type Diterpenes from Euphorbia lagascae. Organic Letters 2007, 9, (3), 489-492.
199.Yang, S. W.; Cordell, G. A., Metabolism Studies of Indole Derivatives Using a Staurosporine Producer, Streptomyces staurosporeus. Journal of Natural Products 1997, 60, (1), 44-48.
200.Asha, K. N.; Chowdhury, R.; Hasan, C. M.; Rashid, M. A., Steroids and polyketides from Uvaria hamiltonii stem bark. Acta Pharmaceutica 2004, 54, (57-63).
201.Wright, J. L. C.; Mcinness, A. G.; Shimizu, S.; Smith, D. G.; Walter, J. A., Identification of C-24 alkyl epimers of marine sterols by 13C nuclear magnetic resonance spectroscopy. Canadian Journal of Chemistry 1978, 56, 1898-1903.
202.Kontiza, I.; Abatis, D.; Malakate, K.; Vagias, C.; Roussis, V., 3-Keto steroids from the marine organisms Dendrophyllia cornigera and Cymodocea nodosa. Steroids 2006, 71, (2), 177-181.
203.Kim, D. S.; Baek, N. l.; Oh, S. R.; Jung, K. Y.; Lee, I. S.; Kim, J. H.; Lee, H. K., Anticomplementary Activity of Ergosterol Peroxide from Naematoloma fasciculare and Reassignment of NMR Data. Archives of Pharmacal Research 1997, 20, (3), 201-205.
204.Kuo, Y. H.; Lo, J. M.; Chan, Y. F., Cytotoxic Components from the Leaves of Schefflera taiwaniana. Journal of the Chinese Chemical Society 2002, 49, (3), 427-431.
205.Kimura, J.; Maki, N., New Loliolide Derivatives from the Brown Alga Undaria pinnatifida. Journal of Natural Products 2002, 65, (1), 57-58.
206.Wu, T. S.; Chang, F. C.; Wu, P. L.; Kuoh, C. S.; Chen, I. S., Constituents of Leaves of Tetradium glabrifolium. Journal of the Chinese Chemical Society 1995, 42, (6), 929-934.
207.Tsai, I. L.; Jeng, Y. F.; Duh, C. Y.; Chen, I. S., Cytotoxic Constituents from the Leaves of Litsea akoensis. The Chinese Pharmaceutical Journal 2001, 53, 291-301.
208.Min, Y. D.; Choi, S. U.; Lee, K. R., Aporphine Alkaloids and their Reversal Activity of Multidrug Resistance (MDR) from the Stems and Rhizomes of Sinomenium acutum. Archives of Pharmacal Research 2006, 29, (8), 627-632.
209.Chiang, Y. M.; Liu, H. K.; Lo, J. M.; Chien, S. C.; Chan, Y. F.; Lee, T. H.; Su, J. K.; Kuo, Y. H., Cytotoxic Constituents of the Leaves of Calocedrus formosana. Journal of the Chinese Chemical Society 2003, 50, (1), 161-166.
210.Wongsinkongman, P.; Brossi, A.; Wang, H. K.; Bastow, K. F.; Lee, K. H., Antitumor Agents. Part 209: Pheophorbide-a Derivatives as Photo-Independent Cytotoxic Agents. Bioorganic & Medicinal Chemistry 2002, 10, (3), 583-591.
211.Cheng, H. H.; Wang, H. K.; Ito, J.; Bastow, K. F.; Tachibana, Y.; Nakanishi, Y.; Xu, Z.; Luo, T. Y.; Lee, K. H., Cytotoxic Pheophorbide-Related Compounds from Clerodendrum calamitosum and C. cyrtophyllum. Journal of Natural Products 2001, 64, (7), 915-919.
212.Gogoi, S.; Argade, N. P., Synthesis of potassium 2,3,4-trihydroxy-2-methylbutanoate: a leaf-closing substance of Leucaena leucocephalam. Tetrahedron 2004, 60, (41), 9093-9093.
213.Kis, K.; Wungsintaweekul, J.; Eisenreich, W.; Zenk, M. H.; Bacher, A., An Efficient Preparation of 2-C-Methyl-d-Erythritol 4-Phosphoric Acid and Its Derivatives. The Journal of Organic Chemistry 2000, 65, (2), 587-592.
214.Fernandes, R. A.; Kumar, P., Enantioselective synthesis of (R)-(-)-mevalonolactone via cyclic sulfate methodology. Tetrahedron: Asymmetry 1999, 10, 4349-4356.
215.Amagata, T.; Usami, Y.; Minoura, K.; Ito, T.; Numata, A., Cytotoxic Substances Produced by a Fungal Strain from a Sponge: Physico-chemical Properties and Structures. The Journal of Antibiotics 1998, 51, (1), 33-40.
216.Eschenmoser, A.; Arigoni, D., Revisited after 50 Years: The 'Stereichemical Interpretation of the Biogenetic Isoprene Rule for the Triterpenes'. Helvetica Chimica Acta 2005, 88, (12), 3011-3050.
217.Afshan, F., Studies on the chemical constituents of the leaves of Azadirachta indica (neem). H.E.J Research Institute Of Chemistry /University Of Karachi 2002, 1-184.

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系統識別號 U0007-2807200911104900
論文名稱(中文) 開發以靈芝廢渣做為雞隻免疫調節飼料之研究
論文名稱(英文) Studies on the develpoment of Ganoderma residues as chicken immunomodulatory feeds
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 97
學期 2
出版年 98
研究生(中文) 林冠伶
學號 M303096013
學位類別 碩士
語文別 中文
口試日期 2009-07-02
論文頁數 106頁
口試委員 委員-林耀輝
委員-劉得任
委員-鄧明中
共同指導教授-商惠芳
指導教授-侯文琪
關鍵字(中) 愛拔益加肉雞
靈芝廢渣
免疫調節
非特異性免疫
腸道菌相
益生菌
關鍵字(英) chick
Ganoderma residues
immunomodulatory
innate immune
intestinal microflora
probiotic bacteria
學科別分類
中文摘要 本研究主要探討將靈芝廢渣 (L) 以及靈芝廢渣之木黴菌固態發酵物 (F) 混合於雞隻飼料中以提升雞隻非特異性免疫之可行性。一日齡愛拔益加肉雞以小雞飼料餵養一週後,將靈芝廢渣及其發酵物以4%和8%之比例混合於小雞飼料後餵養兩週,於雞隻滿三週齡時抽血測其週邊血吞噬活性以及自然殺手細胞活性,以確認先天性免疫之活性,並取糞便檢體測其腸道菌相,另一組為一日齡愛拔益加肉雞以小雞飼料餵養兩週後,將靈芝廢渣及其發酵物以4%和8%之比例混合於中雞飼料後餵養三週,於雞隻滿五週齡時抽血並取糞便檢體。另於體外試驗中,針對RAW 264.7細胞株進行細胞存活性、一氧化氮產生、與脂多醣之結合力以及吞噬活性試驗,並將靈芝廢渣及其發酵物和LGG乳酸桿菌、比菲德菌及大腸桿菌共同培養,看其對菌落增生之影響。體內實驗結果顯示,在吞噬活性方面,於三週齡時8%添加量之實驗組活性優於僅餵食小雞飼料之控制組,而五週齡時,各實驗組之活性皆優於僅餵食中雞飼料之控制組;自然殺手細胞之活性實驗中,三週齡小雞及五週齡小雞於靈芝廢渣發酵物之添加下皆有較好的自然殺手細胞活性。體內腸道菌相檢測中實驗組可提升雞隻腸道內之益生菌數量,但不影響大腸桿菌數量。而在體外試驗中,靈芝廢渣及其發酵物亦可在不造成巨噬細胞毒性之濃度內 (1 mg/mL以下) 有效活化巨噬細胞產生一氧化氮以及增進吞噬活性,並抑制巨噬細胞和脂多醣結合;靈芝廢渣及其發酵物和益生菌於體外共同培養,亦可促進益生菌增生。由此結果顯示,於雞隻飼料中混入靈芝添加物及其發酵物可提升雞隻之非特異性免疫,且β-glucan含量較高之發酵物於自然殺手細胞之毒殺活性較好,可知靈芝廢渣可開發為提升非特異性免疫之雞隻飼養添加,並可進一步測試是否可以取代抗生素之使用,且可對於提高靈芝廢渣之免疫調節指標成分β-glucan做進一步的研究。
英文摘要 Ganoderma is known to have immunomodulatory activities by a series of in vitro and in vivo immune assay. In our study, we focus on the effect of supplemented Ganoderma residues and their fermented products (4% and 8%) on the phagocytosis activities, natural killer cell cytotoxicity and prebiotic effect on intestinal microflora of Arbor Acres chicks. One group was two-week supplemented Ganoderma residues and their fermented products of one-week-old chicks, the other was three-week supplemented of two-week-old chicks. The effect of Ganoderma residues and their fermented products on cell proliferation, NO production, LPS-binding affinity and phagocytosis activities on RAW 264.7 and in vitro probiotic bacteria proliferation were also investigated. The results of in vivo immunomodulatory assay showed that fermented products of Ganoderma residues can enhance natural killer cell cytotoxicity of chicks and both of Ganoderma residues and their fermented products can enhance the phagocytosis activities of chicks. In the in vitro assay, the results showed that Ganoderma residues and their fermented products can enhance NO production, phagocytosis activities and inhibit LPS-binding affinity on RAW 264.7. Ganoderma residues and their fermented products also enhance the probiotic bacteria proliferation in vitro and in vivo assay. In conclusion, Ganoderma residues and their fermented products are effective immunomodulatory products and can enhance the innate immune of chick.
論文目次 中文摘要…………………………………………………………………1
英文摘要…………………………………………………………………3
第一章 前言
壹、靈芝
一、靈芝簡介…………………………………………………4
二、靈芝有效成分介紹………………………………………5
三、靈芝廢渣免疫調節試驗之文獻探討……………………7
貳、免疫系統介紹
一、免疫系統…………………………………………………9
二、一氧化氮與免疫系統…………………………………..10
三、脂多醣與免疫系統……………………………………..11
參、益生菌
一、益生菌…………………………………………………..12
二、益生素…………………………………………………..12
三、益生菌與免疫系統……………………………………..13
肆、雞隻飼料添加物之研究現況…………………………….....14
第二章 實驗動機與目的……………………………………………..15
第三章 實驗材料與方法
第一節 實驗材料與動物………………………………………...17
第二節 靈芝廢渣及其發酵物之體外試驗……………………...18
1.靈芝廢渣及其發酵物之分子量及化學成分檢測………..19
2.靈芝廢渣及其發酵物對RAW 264.7細胞株免疫調節體外試驗之影響………………………………………………..21
3.靈芝廢渣及其發酵物對益生菌增生之影響……………..29
第三節 愛拔益加肉雞餵食靈芝廢渣及其發酵物之體內試驗
1. 餵食流程…………………………………………………..35
2. 愛拔益加肉雞之非特異性免疫活性試驗………………..36
3. 愛拔益加肉雞之腸道菌相檢測…………………………..40
第四節 統計分析………………………………………………...45
第四章 結果與討論
第一節 靈芝廢渣及其發酵物之分子量及化學成分檢測…… 46
第二節 靈芝廢渣及其發酵物對RAW 264.7細胞株免疫調節體外試驗之影響
1.RAW 264.7細胞株之存活率試驗………………………...47
2.RAW 264.7細胞株之一氧化氮含量試驗………………...47
3.RAW 264.7細胞株與脂多醣結合力試驗………………...47
4.RAW 264.7細胞株之吞噬活性試驗……………………...48
討論 靈芝廢渣及其發酵物對RAW 264.7細胞株免疫調節試驗之影響………………………………………………………...49
第三節 靈芝廢渣及其發酵物對益生菌增生之影響
1.靈芝廢渣及其發酵物對LGG乳酸桿菌生長的影響……52
2.靈芝廢渣及其發酵物對比菲德菌生長的影響…………..52
3.靈芝廢渣及其發酵物對大腸桿菌生長的影響…………..52
討論 靈芝廢渣及其發酵物對益生菌增生之影響……………….54
第四節 愛拔益加肉雞餵食靈芝廢渣及其發酵物之體內試驗
1.愛拔益加肉雞餵食過程…………………………………..56
2.愛拔益加肉雞體重紀錄…………………………………..56
3.愛拔益加肉雞週邊血液白血球吞噬能力………………..56
4.愛拔益加肉雞自然殺手細胞活性………………………..56
5.愛拔益加肉雞腸道菌相檢測:乳酸桿菌………………….57
6.愛拔益加肉雞腸道菌相檢測:比菲德菌………………….57
7.愛拔益加肉雞腸道菌相檢測:大腸桿菌………………….58
討論一 靈芝廢渣及其發酵物對愛拔益加肉雞非特異性免疫之影響…………………………………………………………...59
討論二 靈芝廢渣及其發酵物對愛拔益加肉雞腸道菌相之影響
…………………………………………………………….61
結論……………………………………………………………………..62
參考資料………………………………………………………………..63
實驗結果圖表…………………………………………………………..66
附錄……………………………………………………………………..86
參考文獻 1.Su, C.H., et al., Fungal mycelia as the source of chitin and polysaccharides and their applications as skin substitutes. Biomaterials, 1997. 18(17): p. 1169-74.
2.Gao, Y., et al., Ganoderma lucidum polysaccharide fractions accelerate healing of acetic acid-induced ulcers in rats. J Med Food, 2004. 7(4): p. 417-21.
3.Sliva, D., Ganoderma lucidum (Reishi) in cancer treatment. Integr Cancer Ther, 2003. 2(4): p. 358-64.
4.Shao, B.M., et al., Immune receptors for polysaccharides from Ganoderma lucidum. Biochem Biophys Res Commun, 2004. 323(1): p. 133-41.
5.Wang, G., et al., Antitumor active polysaccharides from the Chinese mushroom Songshan lingzhi, the fruiting body of Ganoderma tsugae. Biosci Biotechnol Biochem, 1993. 57(6): p. 894-900.
6.Wasser, S.P. and A.L. Weis, Therapeutic effects of substances occurring in higher Basidiomycetes mushrooms: a modern perspective. Crit Rev Immunol, 1999. 19(1): p. 65-96.
7.Lin, Z.B. and H.N. Zhang, Anti-tumor and immunoregulatory activities of Ganoderma lucidum and its possible mechanisms. Acta Pharmacol Sin, 2004. 25(11): p. 1387-95.
8.Shieh, Y.H., et al., Evaluation of the hepatic and renal-protective effects of Ganoderma lucidum in mice. Am J Chin Med, 2001. 29(3-4): p. 501-7.
9.Su, C.H., et al., High-performance liquid chromatographic analysis for the characterization of triterpenoids from Ganoderma. J Chromatogr Sci, 2001. 39(3): p. 93-100.
10.Liu, X., et al., Antitumor activity of the sporoderm-broken germinating spores of Ganoderma lucidum. Cancer Lett, 2002. 182(2): p. 155-61.
11.Kino, K., et al., Isolation and characterization of a new immunomodulatory protein, ling zhi-8 (LZ-8), from Ganoderma lucidium. J Biol Chem, 1989. 264(1): p. 472-8.
12.Chiesa, M.D., et al., Refocusing of B-cell responses following a single amino acid substitution in an antigen. Immunology, 2001. 103(2): p. 172-8.
13.Girotti, M., et al., Cytosolic phospholipase A2 translocates to forming phagosomes during phagocytosis of zymosan in macrophages. J Biol Chem, 2004. 279(18): p. 19113-21.
14.Zhu, L., C. Gunn, and J.S. Beckman, Bactericidal activity of peroxynitrite. Arch Biochem Biophys, 1992. 298(2): p. 452-7.
15.Schumann, R.R., et al., Structure and function of lipopolysaccharide binding protein. Science, 1990. 249(4975): p. 1429-31.
16.Kitchens, R.L., Role of CD14 in cellular recognition of bacterial lipopolysaccharides. Chem Immunol, 2000. 74: p. 61-82.
17.Fuller, R., Probiotics in man and animals. J Appl Bacteriol, 1989. 66(5): p. 365-78.
18.Hamilton-Miller, J.M., The role of probiotics in the treatment and prevention of Helicobacter pylori infection. Int J Antimicrob Agents, 2003. 22(4): p. 360-6.
19.Wollowski, I., G. Rechkemmer, and B.L. Pool-Zobel, Protective role of probiotics and prebiotics in colon cancer. Am J Clin Nutr, 2001. 73(2 Suppl): p. 451S-455S.
20.Sanders, M.E., Considerations for use of probiotic bacteria to modulate human health. J Nutr, 2000. 130(2S Suppl): p. 384S-390S.
21.Niedzielin, K., H. Kordecki, and B. Birkenfeld, A controlled, double-blind, randomized study on the efficacy of Lactobacillus plantarum 299V in patients with irritable bowel syndrome. Eur J Gastroenterol Hepatol, 2001. 13(10): p. 1143-7.
22.Gibson, G.R., et al., Selective stimulation of bifidobacteria in the human colon by oligofructose and inulin. Gastroenterology, 1995. 108(4): p. 975-82.
23.Erickson, K.L. and N.E. Hubbard, Probiotic immunomodulation in health and disease. J Nutr, 2000. 130(2S Suppl): p. 403S-409S.
24.Vaarala, O., Immunological effects of probiotics with special reference to lactobacilli. Clin Exp Allergy, 2003. 33(12): p. 1634-40.
25.Liu, Y.W., et al., Immunomodulatory activity of dioscorin, the storage protein of yam (Dioscorea alata cv. Tainong No. 1) tuber. Food Chem Toxicol, 2007. 45(11): p. 2312-8.
26.Misko, T.P., et al., A fluorometric assay for the measurement of nitrite in biological samples. Anal Biochem, 1993. 214(1): p. 11-6.
27.Carracedo, J., et al., The effect of LPS, uraemia, and haemodialysis membrane exposure on CD14 expression in mononuclear cells and its relation to apoptosis. Nephrol Dial Transplant, 2002. 17(3): p. 428-34.
28.Gamal-Eldeen, A.M., et al., Chemically-modified polysaccharide extract derived from Leucaena leucocephala alters Raw 264.7 murine macrophage functions. Int Immunopharmacol, 2007. 7(6): p. 871-8.
29.Foukas, L.C., et al., Phagocytosis of Escherichia coli by insect hemocytes requires both activation of the Ras/mitogen-activated protein kinase signal transduction pathway for attachment and beta3 integrin for internalization. J Biol Chem, 1998. 273(24): p. 14813-8.
30.Okazaki, W., et al., Induction of lymphoid leukosis transplant able tumours and the establishment of lymphoblastoid cell lines. Avian Pathol, 1980. 9(3): p. 311-29.
31.Kushima, K., et al., Flow cytometric analysis of chicken NK activity and its use on the effect of restraint stress. J Vet Med Sci, 2003. 65(9): p. 995-1000.
32.Green, L.C., et al., Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem, 1982. 126(1): p. 131-8.
33.Gibson, G.R. and M.B. Roberfroid, Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J Nutr, 1995. 125(6): p. 1401-12.
34.Roberfroid, M.B., et al., Colonic microflora: nutrition and health. Summary and conclusions of an International Life Sciences Institute (ILSI) [Europe] workshop held in Barcelona, Spain. Nutr Rev, 1995. 53(5): p. 127-30.
35.O'Mahony, L., et al., Probiotic impact on microbial flora, inflammation and tumour development in IL-10 knockout mice. Aliment Pharmacol Ther, 2001. 15(8): p. 1219-25.
36.Su, C.H., et al., Use of Ganoderma tsugae for the treatment of Human Chronic Skin Ulcers. Mushroom Science, 2004. XVI. 659-662.
37.林育審 飼糧添加黃耆、淫羊藿及補骨脂對蛋雞產蛋性狀與免疫反應之影響 2008



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系統識別號 U0007-2907200914041300
論文名稱(中文) 天然植物資源對人類神經膠質瘤細胞抑制之研究
論文名稱(英文) Inhibitory Effects of Natural Botany Resources on Human Glioma Cells
校院名稱 臺北醫學大學
系所名稱(中) 生藥學研究所
系所名稱(英) Graduate Institute of Pharmacognosy
學年度 97
學期 2
出版年 98
研究生(中文) 尤郁雯
學號 M303096010
學位類別 碩士
語文別 中文
口試日期 2009-07-14
論文頁數 100頁
口試委員 委員-張睌E
委員-許金玉
委員-蔣永孝
委員-陳彥州
指導教授-楊玲玲
關鍵字(中) 人類神經膠質瘤
細胞毒性
抗氧化
自由基
脂質過氧化
關鍵字(英) human malignant glioma
cytotoxicity
antioxidant
free radical
lipid peroxidation
學科別分類
中文摘要 神經膠質瘤在原發性腦瘤中最為常見,其中有三分之二屬惡性。美國每年約有17,000 名被診斷出患有原發性腦瘤,而神經膠質瘤就佔 60 %。根據世界衛生組織分類為高惡性度屬於第三級退化性星狀細胞 (anaplastic astrocytoma, AA) 及第四級多型性神經膠質母細胞瘤 (glioblastoma multiforme, GBM) 最為普遍。腦瘤治療通常以手術切除為主,輔以化學及放射線治療。但腦瘤治療的預後不佳,特別是化學及放射線治療會產生游離自由基,不僅會殺死癌細胞,連正常細胞也會殺死。最近有研究指出,抗氧化劑能提高治療效果、降低副作用並保護正常細胞。天然植物資源豐富種類繁多,據科學證實其具保健並防止癌症在復發之潛力,且多酚具有預防癌症之效果。
本研究共採集 60 種天然植物資源製備成冷凍乾燥甲醇取物,利用人類神經膠質腦瘤細胞屬於第三級的 U-87 MG 與第四期的 GBM 8401 以 MTT 還原能力法來評估萃取物對於神經膠質瘤細胞抑制生長之作用進行篩選。其中以 YY-02 甲醇萃取物抑制率最高,接著再以不同極性之溶劑正己烷、乙酸乙酯、丙酮及甲醇依序進行分離萃取,再評估其總多酚及類黃酮之含量、抗氧化及抗脂質過氧化能力,結果以甲醇再分離萃取的效果最好,並且對此兩種人類惡化神經膠質瘤細胞均具有抑制效果並具有濃度依存性,另由細胞形態及細胞週期分析結果亦發現 YY-02 甲醇分離萃取物之抗腫瘤機制乃經由細胞凋亡之模式造成神經膠質瘤細胞死亡,因此 YY-02 將來可能成為一具有開發潛力的藥物以做為治療神經膠質瘤之新應用。
英文摘要 Gliomas are the most common type of primary brain tumor. Nearly two-thirds of gliomas are highly malignant lesions that account for a disproportionate shape of brain tumor-related morbidity and mortality. Approximately 17,000 primary brain tumors are diagnosed every year, and of those, about 60% are gliomas. Anaplastic astrocytoma (AA, grade III) and glioblastoma multiforme (GBM, grade IV), the high grade gliomas, are the most common type hence we employed the human malignant glioma cell lines of U87 MG (III) and GBM 8401 (IV) as target to examined antiproliferation activity by MTT reduction.
Brain tumor therapy usually gets surgical resection for the first priority and followed by radiotherapy and chemotherapy. All brain tumor therapies now usually result in the poor prognosis. Especially, radiotherapy and chemotherapy would generate free radicals, kill not only cancer cells but also healthy cells. Recent studies have demonstrated that antioxidant could enhance the therapeutic effects, decrease the side effects and protect normal tissue damage. Phytochemicals are plant-derived compounds under scientific research for potential health promoting properties and recurrence of cancer. Epidemiological and preclinical evidence suggests that polyphenolic phytochemicals possess cancer chemopreventive properties.
In this study, a high through screening method was established to investigate 60 kinds of natural botany resource on anti-brain tumor activity. YY-02 methanol extract (ME) is the most efficacy of antiproliferation on human malignant glimoas. Furthermore, ME was fractionated with n-hexane (HeF), ethyl acetate (EA), acetone (AcF) and methanol (MeF). Outcomes of total polyphenol, flavonoid content, and DPPH radical scavenged, chelated ferrous ion, lipid peroxidation (LPO) inhibition of liver, kidney and brain tissue measurement, that exerted the MeF is a highest potential fraction in YY-02. Otherwise, view of morphological assay and cell cycle analysis, YY-02 exhibited the cell death pathway by apoptosis. In conclusion, YY-02 is a phytochemical of chemotherapy agents on brain tumor.
論文目次 前言-------------------------------------------------------1
材料及儀器-------------------------------------------------9
一、試藥與試劑---------------------------------------------9
二、實驗用儀器--------------------------------------------14
三、樣品萃取與分離----------------------------------------15
四、細胞株------------------------------------------------17
五、試液之配製--------------------------------------------18
實驗方法--------------------------------------------------23
一、細胞株之繼代培養--------------------------------------23
二、細胞凍存----------------------------------------------23
三、冷凍細胞之活化----------------------------------------23
四、腦細胞初代培養----------------------------------------24
五、對正常腦細胞及人類神經膠質瘤細胞之增生試驗------------24
六、細胞之鏡檢反應分析------------------------------------26
七、細胞週期分析------------------------------------------26
八、總多酚量之測定----------------------------------------27
九、總黃酮類量之測定--------------------------------------27
十、抗氧化活性評估----------------------------------------28
1.清除 DPPH 自由基能力之測定------------------------------28
2.還原能力之測定(ferric reducing antioxidant power, FRAP)-29
3.螯合亞鐵離子 (ferrous ion, Fe2+ ) 能力之測定------------30
4.清除超氧陰離子 (superoxide radical, O2• -) 能力之測定--31
5.抑制黃嘌呤氧化酶 (XOD) 活性之檢測-----------------------32
6.脂質過氧化 (lipid peroxidation, LPO) 活性---------------33
十一、統計分析--------------------------------------------38
結果------------------------------------------------------39
一、60種台灣原生種植物對人類腦瘤及正常腦細胞之毒性結果----39
二、YY-02 誘導人類腦瘤細胞凋亡----------------------------45
三、YY-02 總多酚及黃酮類之含量----------------------------54
四、YY-02 自由基清除及抗氧化能力之測定結果----------------57
1.清除 DPPH 自由基之能力----------------------------------57
2.螯合亞鐵離子之能力--------------------------------------59
3.還原能力 (FRAP)-----------------------------------------61
4.清除超氧陰離子之能力------------------------------------62
5.抑制 XOD 之能力-----------------------------------------63
6.對於腦均質液,YY-02 能抑制 Fe(NH4)2(SO4)2.6H2O 誘導
所產生之脂質過氧化,但本身卻不誘導脂質過氧化------------64
7.抑制 FeCl2 誘導肝/腎粒線體脂質過氧化之能力--------------67
綜合以上結果----------------------------------------------72
討論------------------------------------------------------73
參考文獻--------------------------------------------------78
附錄------------------------------------------------------89
參考文獻 Afonso V, Champy R, Mitrovic D, Collin P, Lomri A. 2007. Reactive oxygen species and superoxide dismutases: role in joint diseases. Joint Bone Spine., 74(4):324-329.
Agarwal A, Sharma V, Tewari R, Koul N, Josheph C and Sen E. 2008. Epigallocatechin-3-gallate exhibits anti-tumor effect by perturbing redox homeostasis, modulating the release of pro-inflammatory mediators and decreasing the invasiveness of glioblastoma cells. Mol Med Rep., 1:511-515.
Alley MC, Scudiero DA, Monks A, Hursey ML, Czerwinski MJ, Fine DL, Abbott BJ, Mayo JG, Shoemaker RH, Boyd MR. 1988. Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay. Cancer Res., 48(3):589-601.
Anandakumar PP, Malarkodi SP, Sivaprasad TR, Saravanan GD. 2007. Antioxidant DL-alpha lipoic acid as an attenuator of adriamycin induced hepatotoxicity in rat model. Indian J Exp Biol., 45(12):1045-1049.
Balachandar AV, Malarkodi KP, Varalakshmi P. 2003. Protective role of DLalpha-lipoic acid against adriamycin-induced cardiac lipid peroxidation. Hum Exp Toxicol., 22(5):249-254.
Benzie IF, Strain JJ. 1996. The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": the FRAP assay. Anal Biochem., 239(1):70-76.
Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem., 72:248-254.
Chen HJ. 2004. Effect of EGCG on proliferation inhibition and apoptotic mechanism of human brain tumor cells.
Cheng YL, Chang WL, Lee SC, Liu YG, Chen CJ, Lin SZ, Tsai NM, Yu DS, Yen CY, Harn HJ. 2004. Acetone extract of Angelica sinensis inhibits proliferation of human cancer cells via inducing cell cycle arrest and apoptosis. Life Sci., 75(13):1579-1594.
CTEP. 1998. Cancer Therapy Evaluation Program (CTEP), Division of Cancer Treatment and Diagnosis (DCTD), NCI.
Dario A, Tomei G. 2006. The safety of the temozolomide in patients with malignant glioma. Curr Drug Saf., 1(2):205-222.
Dinis TC, Madeira VM, Almeida LM. 1994. Action of phenolic derivatives (acetaminophen, salicylate, and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Arch of Biochem Biophys., 315(1):161-169.
Earnshaw WC, Martins LM, Kaufmann SH. 1999. Mammalian caspases: structure, activation, substrates, and functions during apoptosis. Annu Rev Biochem., 68:383-424.
Elmore S. 2007. Apoptosis: A Review of Programmed Cell Death. Toxicol Pathol., 35(4):495-516.
Eric R. Kandel, James H. Schwartz, Thomas M. Jessell. 2000. Principles of Neural Science, 4th Edition, McGraw-Hill Medical Companies.
Engel RH, Evens AM. 2006. Oxidative stress and apoptosis: a new treatment paradigm in cancer. Front in Biosci., 11:300-312.
Ferguson PJ, Kurowska E, Freeman DJ, Chambers AF, Koropatnick DJ. 2004. A flavonoid fraction from cranberry extract inhibits proliferation of human tumor cell lines. J Nutr., 134(6):1529-1535.
Ferguson PJ, Kurowska EM, Freeman DJ, Chambers AF, Koropatnick J. 2006. In vivo inhibition of growth of human tumor lines by flavonoid fractions from cranberry extract. Nutr Cancer., 56(1):86-94.
Fuster JJ, Sanz-González SM, Moll UM, Andrés V. 2007. Classic and novel roles of p53: prospects for anticancer therapy. Trends Mol Med., 13(5):192-199.
George S, Brat P, Alter P, Amiot MJ. 2005. Rapid Determination of Polyphenols and Vitamin C in Plant-Derived Products. J. Agric. Food Chem., 53:1370-1373.
Gogvadze V, Orrenius S, Zhivotovsky B. 2009. Mitochondria as targets for chemotherapy. Apoptosis., 14(4):624-640.
Goodlett CR, Horn KH. 2001. Mechanisms of alcohol-induced damage to the developing nervous system. Alcohol Res Health., 25(3):175-184.
Gupta S, Agrawal A, Agrawal S, Su H, Gollapudi S. 2006. A paradox of immunodeficiency and inflammation in human aging: lessons learned from apoptosis. Immun Ageing., 19;3:5.
Huerta S, Goulet EJ, Huerta-Yepez S, Livingston EH. 2007. Screening and detection of apoptosis. J Surg Res., 139(1):143-156.
Joshi G, Sultana R, Tangpong J, Cole MP, St Clair DK, Vore M, Estus S, Butterfield DA. 2005. Free radical mediated oxidative stress and toxic side effects in brain induced by the anti cancer drug adriamycin: insight into chemobrain. Free Radic Res., 39(11):1147-1154.
Kalivendi SV, Konorev EA, Cunningham S, Vanamala SK, Kaji EH, Joseph J, Kalyanaraman B. 2005. Doxorubicin activates nuclear factor of activated T-lymphocytes and Fas ligand transcription: role of mitochondrial reactive oxygen species and calcium. Biochem J., 389(Pt 2):527-539.
Khan S, Ramwani JJ, O'Brien PJ. 1992. Hepatocyte toxicity of mechlorethamine and other alkylating anticancer drugs. Role of lipid peroxidation. Biochem Pharmacol., 43(9):1963-1967.
Kintzel PE. 2001. Anticancer drug-induced kidney disorders. Drug Saf., 24(1):19-38.
Kong LD, Cai YHuang WW, Cheng CH and Tan RX. 2000. Inhibition of xanthine oxidase by some Chinese medicinal plants used to treat gout. J Ethnopharmacol., 73(1-2):199-207.
Koren G, Beatty K, Seto A, Einarson TR, Lishner M. 1992. The effects of impaired liver function on the elimination of antineoplastic agents. Ann Pharmacother., 26(3):363-371.
Koukourakis GV, Kouloulias V, Zacharias G, Papadimitriou C, Pantelakos P, Maravelis G, Fotineas A, Beli I, Chaldeopoulos D, Kouvaris J. 2009. Temozolomide with radiation therapy in high grade brain gliomas: pharmaceuticals considerations and efficacy; a review article. Molecules., 14(4):1561-1577.
Kreisl TN. 2009. Chemotherapy for Malignant Gliomas. Semin Radiat Oncol., 19(3):150-154.
La Rocca RV, Mehdorn HM. 2009. Localized BCNU chemotherapy and the multimodal management of malignant glioma. Curr Med Res Opin., 25(1):149-160.
Lamaison JLC and Carnet A. 1990. Teneurs en principaux flavonoids des fleurs de Crataegeus monogyna Jacq et de Crataegeus laevigata (Poiret D. C) en fonction de la vegetation. Pharm Acta Helv., 65: 315-320.
Langley B, Ratan RR. 2004. Oxidative stress-induced death in the nervous system: cell cycle dependent or independent? J Neurosci Res., 77(5):621-629.
Lee HH, Lin CT, Yang LL. 2007. Neuroprotection and free radical scavenging effects of Osmanthus fragrans. J Biomed Sci., 14(6):819-827.
Lee WH, Jin JS, Tsai WC, Chen YT, Chang WL, Yao CW, Sheu LF, Chen A. 2006. Biological inhibitory effects of the Chinese herb danggui on brain astrocytoma. Pathobiology., 73(3):141-148.
Lin J, Chen LY, Lin ZX, Zhao ML. 2007. The effect of triptolide on apoptosis of glioblastoma multiforme (GBM) cells. J Int Med Res., 35(5):637-643.
Lin PC, Chen YL, Chiu SC, Yu YL, Chen SP, Chien MH, Chen KY, Chang WL, Lin SZ, Chiou TW, Harn HJ. 2008. Orphan nuclear receptor, Nurr-77 was a possible target gene of butylidenephthalide chemotherapy on glioblastoma multiform brain tumor. J Neurochem., 106(3):1017-1026.
Louis DN. 2006. Molecular Pathology of Malignant Gliomas. Annu Rev Pathol Mech Dis., 1:97-117
Injac R, Strukelj B. 2008. Recent advances in protection against doxorubicin-induced toxicity. Technol Cancer Res Treat., 7(6):497- 516.
Magalhães LM, Segundo MA, Reis S, Lima JL. 2008. Methodological aspects about in vitro evaluation of antioxidant properties. Anal Chim Acta., 613(1):1-19.
Majno G, Joris I. 1995. Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol., 146(1): 3-15.
Malarkodi KP, Balachandar AV, Varalakshmi P. 2003. Protective effect of lipoic acid on adriamycin induced lipid peroxidation in rat kidney. Mol Cell Biochem., 247(1-2):9-13.
Mann CD, Neal CP, Garcea G, Manson MM, Dennison AR, Berry DP. 2009. Phytochemicals as potential chemopreventive and chemotherapeutic agents in hepatocarcinogenesis. Eur J Cancer Prev., 18(1):13-25.
Martin, S. J. and Green, D. R. 1995. Apoptosis and cancer: the failure of controls on cell death and cell survival. Crit Rev Oncol Hematol., 18(2):137-153.
Mastrangelo, A. J. and Betenbaugh, M. J. 1988. Overcoming apoptosis: new methods for improving protein-expression systems. Trends Biotechnol., 16(2):88-95.
McConkey D. J. 1998. Biochemical determinants of apoptosis and necrosis. Toxicol Lett., 99(3):157-168.
Mizutani H, Tada-Oikawa S, Hiraku Y, Kojima M, Kawanishi S. 2005. Mechanism of apoptosis induced by doxorubicin through the generation of hydrogen peroxide. Life Sci., 76(13):1439-1453.
Ogura M. 2001. Adriamycin (doxorubicin). Gan To Kagaku Ryoho., 28(10):1331-1338.
Oltvai ZN, Milliman CL, Korsmeyer SJ. 1993. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell., 74(4):609-619.
Oz E, Ilhan MN. 2006. Effects of melatonin in reducing the toxic effects of doxorubicin. Mol Cell Biochem., 286(1-2):11-15.
Pan MH, Ghai G, Ho CT. 2008. Food bioactives, apoptosis, and cancer. Mol Nutr Food Res., 52(1):43-52.
Singh RP, Sharad S, Kapur S. 2004. Free Radicals and Oxidative Stress in Neurodegenerative Diseases: Relevance of Dietary Antioxidants. J Indian Acad Clin Med., 5(3):218-225.
Reardon DA, Wen PY. 2006. Therapeutic advances in the treatment of glioblastoma: rationale and potential role of targeted agents. Oncologist., 11(2):152-164.
Reed JC, Pellecchia M. 2005. Apoptosis-based therapies for hematologic malignancies. Blood., 106(2):408-418.
Ren W, Qiao Z, Wang H, Zhu L, Zhang L. 2003. Flavonoids: promising anticancer agents. Med Res Rev., 23(4):519-34.
Robak J, Gryglewski RJ. 1988. Flavonoids are scavengers of superoxide anions. Biochem Pharmacol., 37(5):837-841.
Roberto LS. 2008. Organic acids influence on DPPH_ scavenging by ascorbic acid. Food Chem., 107:40-43.
Sathornsumetee S and Rich JN. 2008. Designer Therapies for Glioblastoma Multiforme. Ann N Y Acad Sci., 1142:108-132.
Stan AC, Casares S, Radu D, Walter GF, Brumeanu TD. 1999. Doxorubicininduced cell death in highly invasive human gliomas. Anticancer Res., 19(2A):941-950.
See SJ, Gilbert MR. 2007. Chemotherapy in Adults with Gliomas. Ann Acad Med Singapore., 36(5):364-369.
Shen SC, Ko CH, Tseng SW, Tsai SH, Chen YC. 2004. Structurally related antitumor effects of flavanones in vitro and in vivo: involvement of caspase 3 activation, p21 gene expression, and reactive oxygen species production. Toxicol Appl Pharmacol., 197:84-95.
Shimada K, Fujikawa K, Yahara K, Nakamura T. 1992. Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. J Agric Food Chem., 40: 945-948.
Shimizu T, O'Connor PM, Kohn KW, Pommier Y. 1995. Unscheduled activation of cyclin B1/Cdc2 kinase in human promyelocytic leukemia cell line HL60 cells undergoing apoptosis induced by DNA damage. Cancer Res., 55(2):228-231.
Simone CB 2nd, Simone NL, Simone V, Simone CB. 2007. Antioxidants and other nutrients do not interfere with chemotherapy or radiation therapy and can increase kill and increase survival, part 1. Altern Ther Health Med., 13(1):22-28.
Simone CB 2nd, Simone NL, Simone V, Simone CB. 2007. Antioxidants and other nutrients do not interfere with chemotherapy or radiation therapy and can increase kill and increase survival, part 2. Altern Ther Health Med., 13(2):40-47.
Sulman EP, Guerrero M, Aldape K. 2009. Beyond grade: molecular pathology of malignant gliomas. Semin Radiat Oncol., 19(3):142- 149.
Tsai NM, Lin SZ, Lee CC, Chen SP, Su HC, Chang WL, Harn HJ. 2005. The antitumor effects of Angelica sinensis on malignant brain tumors in vitro and in vivo. Clin Cancer Res., 11(9):3475-3484.
Tsai NM, Chen YL, Lee CC, Lin PC, Cheng YL, Chang WL, Lin SZ, Harn HJ. 2006. The natural compound n-butylidenephthalide derived from Angelica sinensis inhibits malignant brain tumor growth in vitro and in vivo. J Neurochem., 99(4):1251-1262.
Valko M, Rhodes CJ, Moncol J, Izakovic M, Mazur M. 2006. Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chem Biol Interact., 160(1):1-40.
Voss C, Eyol E, Berger MR. 2006. Identification of potent anticancer activity in Ximenia americana aqueous extracts used by African traditional medicine. Toxicol Appl Pharmacol., 211(3):177-187.
Wajant H. 2002. The Fas signaling pathway: more than a paradigm. Science., 296(5573):1635-1636.
Wen PY, Kesari S. 2008. Malignant gliomas in adults. N Engl J Med., 359(5):492-507.
Wong SH, Knight JA, Hopfer SM, Zaharia O, Leach CN. Jr, Sunderman FW Jr. 1987. Lipoperoxides in plasma as measured by liquid- chromatographic separation of malondialdehyde-thiobarbituric acid adduct. Clin Chem., 33(2 Pt 1):214-220.
Wu J, Wu Y, Yang BB. 2002. Anticancer activity of Hemsleya amabilis extract. Life Sci., 71(18):2161-2170.
Yin D, Wakimoto N, Xing H, Lu D, Huynh T, Wang X, Black KL, Koeffler HP. 2008. Cucurbitacin B markedly inhibits growth and rapidly affects the cytoskeleton in glioblastoma multiforme. Int J Cancer., 123(6):1364-1375.

 


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