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系統識別號 U0007-2807201118451400
論文名稱(中文) 臺灣特有種植物之抗光老化活性系列探討—山枇杷
論文名稱(英文) A series of studies on the anti-photoaging activity in Taiwanese endemic plants—Eriobotrya deflexa
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 99
學期 2
出版年 100
研究生(中文) 郭香君
研究生(英文) Hsiang-Chun Kuo
學號 M303098008
學位類別 碩士
語文別 中文
口試日期 2011-07-08
論文頁數 147頁
口試委員 委員-郭悅雄
委員-張溫良
委員-徐鳳麟
委員-侯文琪
指導教授-李美賢
中文關鍵字 山枇杷  人類皮膚纖維母細胞  抗光老化 
英文關鍵字 Eriobotrya deflexa  human dermal fibroblasts  photoaging 
學科別分類
中文摘要 山枇杷 (Eriobotrya deflexa (Hemsl.) Nakai) 薔薇科 (Rosaceae)枇杷屬 (Eriobotrya) 常綠喬木,為台灣特有種植物,分布於台灣全島低、中海拔闊葉林中。本論文乃將山枇杷葉以 95% 乙醇萃取,分別以正己烷、乙酸乙酯、正丁醇及水進行部份劃分,其中乙酸乙酯層及正丁醇層萃取物,經由各種管柱層析方法進行分離純化,於乙酸乙酯層得到 5 個化合物,正丁醇層得到 8 個化合物,經理化及光譜相關數據與參考文獻比對後,確認構造為 7 個 flavonol glycosides 化合物:(1)、(2)、(3)、(4)、(5)、(6) 及 (9) ﹔1 個 flavone 化合物:(10) ﹔以及 2 個 caffeoyl 衍生之化合物:(7) 及 (8),其中有 3 個 flavonol glycosides 化合物:(3)、(5) 及 (8) 同時存在於兩層萃取物中。進而以高效液相層析-固相萃取-核磁共振 (LC-SPE-NMR) 進行分析,於乙酸乙酯層分析出含有 5 個 flavonol glycosides:(2)、(3)、(5)、(6)和 (9) 以及 5 個含有 p-coumaryl 基團化合物 ﹔於正丁醇層經管柱層析前處理,分析出含有 1 個 flavonol glycosides 化合物:(9)、1 個 caffeoyl 衍生之化合物、2 個含有 p-coumaryl 基團 及 2 個含有 caffeoyl 基團的化合物。進而以人類皮膚纖維母細胞,經紫外線 B 照射來誘導細胞大量分泌基質金屬蛋白酶 (MMPs),利用 MMP-1 做為抗光老化活性指標。結果顯示:在低細胞毒性下,化合物 (1)、(2)、(5) 及 (7) 於 100 μM 時,具有較明顯之抑制作用。
英文摘要 Eriobotrya deflexa (Hemsl.) Nakai (Rosaceae) is an evergreen tree and endemic throughout of Taiwan. The leaves of E. deflexa were extracted with 95% ethanol, then partitioned with a sequence of n-hexane, ethyl acetate (EtOAc), and n-butanol (n-BuOH). The EtOAc layer and n-BuOH layer were passed by a series of chromatographic analysis, isolated, and purified. The structures of isolated compounds were identified by various physical and spectroscopic characterizations. There were five compounds obtained from the EtOAc layer and eight compounds from the n-BuOH layer, but three compounds were in both layers at the same time. Totally, ten compounds were obtained from E. deflexa, including seven flavonol glycosides: (1), (2), (3), (4), (5), (6), (9), one flavone: (10), and two caffeoyl derivatives: (7), (8) in the present study. By using high-performance liquid chromatography with postcolumn solid-phase extraction to nuclear magnetic resonance spectroscopy (LC-SPE-NMR) to analyse the constituents of EtOAc layer and fraction 3 of n-BuOH layer from E. deflexa. There were five flavonol glycosides: (2), (3), (5), (6), (9) and five compounds with p-coumaryl group in EtOAc layer, and there are one flavonol glycoside: (9), one caffeoyl derivative, two compounds with p-coumaryl group and two compounds with caffeoyl group in fraction 3 of n-BuOH layer with LC-SPE-NMR analysis. The isolated compounds were evaluated the anti-photoaging activity using MMP-1 as a target on the production of MMP-1 secretions after
XV
UVB irradiation in WS-1. The results showed that compounds (1), (2), (5) and (7) (100 μM) exhibited the MMP-1 inhibitory effects at low cytotoxicity in WS-1 cells.
論文目次 目錄 I
圖目錄 V
表目錄 IX
縮寫表 XII
摘要 XIII
Abstract XIV
壹、緒論 1
一、 山枇杷基本介紹及其同屬植物成分研究回顧 1
(一) 山枇杷基本介紹 1
(二) 枇杷屬植物相關介紹 2
(三) 枇杷屬植物活性研究回顧 7
(四) 枇杷屬植物之化學成分研究 8
二、 LC-SPE-NMR 之介紹 15
三、 研究背景 17
(一) 皮膚結構介紹 17
(二) 紫外線對皮膚影響之介紹 20
(三) 基質金屬蛋白酶之介紹 21
(四) 膠原蛋白 22
(五) 光老化之機制 24
四、 研究動機 26
貳、實驗材料與方法 27
一、 山枇杷之萃取與分離 27
(一) 試藥及溶媒 27
(二) 色層分析法材料 27
(三) 儀器 28
(四) LC-SPE-NMR 技術之儀器設備 29
(五) 萃取及分離流程 30
(六) 水解及單醣的組成分析 35
(七) LC-SPE-NMR分析 36
(八) 各成分之物理數據 38
二、 山枇杷之活性實驗 46
(一) 藥品與試劑 46
(二) 細胞株及細胞培養液 46
(三) 儀器 47
(四) 分析軟體 47
(五) WS-1細胞株及細胞培養 47
(六) WS-1細胞株存活率試驗 48
(七) MMP-1 酵素活性染色試驗 48

參、實驗結果 50
一、 山枇杷之化合物及其結構鑑定 50
(一) Compound (1) 之結構解析 50
(二) Compound (2) 之結構解析 54
(三) Compound (3) 之結構解析 58
(四) Compound (4) 之結構解析 62
(五) Compound (5) 之結構解析 66
(六) Compound (6) 之結構解析 70
(七) Compound (7) 之結構解析 74
(八) Compound (8) 之結構解析 77
(九) Compound (9) 之結構解析 80
(十) Compound (10) 之結構解析 84
二、 LC-SPE-NMR之化合物分析 87
(一) 山枇杷之乙酸乙酯層分析 87
(二) 山枇杷之正丁醇層部分分析 90
三、 山枇杷葉之活性成分探討 92
(一) 人類皮膚纖維母 (WS-1) 之細胞存活率試驗 92
(1) 山枇杷四層劃分部分於人類皮膚纖維母之細胞存活率試驗 92
(2) 山枇杷分離之成分於人類皮膚纖維母之細胞存活率試驗 93

(二) 人類皮膚纖維母細胞之基質金屬蛋白酶-1 (MMP-1) 活性染色試驗 (Zymography) 93
(1) 山枇杷四層劃分部分於人類皮膚纖維母細胞之MMP-1活性染色試驗 93
(2) 山枇杷分離之成分於人類皮膚纖維母之MMP-1 活性染色試驗 93
肆、討論 95
伍、結論 101
參考文獻 104
附圖 112

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