九節茶葉部促進U937細胞株吞噬作用之研究

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Title:	九節茶葉部促進U937細胞株吞噬作用之研究 Studies on enhancing U937 cells phagocytosis from the leaves of Chloranthus glabra
Authors:	李美璇 Mei-hsuan Lee
Contributors:	黃秀琴嘉南藥理科技大學：藥物科技研究所
Keywords:	九節茶葉 U937細胞株毒性試驗標示綠色螢光蛋白之克雷白氏肺炎桿菌促進 U937 細胞吞噬作用有效成分分離 isolate effective constituents enhancing phagocytosis of U937 cells leaves of Chloranthus glaber U937 cell line cytotoxicity test Klebsiella pneumoniae-gfp
Date:	2007
Issue Date:	2010-06-01 09:21:32 (UTC+8)
Abstract:	本論文目的主要建立增強免疫弁鄐尿z選平台，進行台灣草藥有效成分之分離。本研究選擇台灣產之九節茶葉及莖部 (CGL，CGS)、黃藤 (DM)、山澤蘭 (EF) 及龍葵 (SN) 等四種草藥，分別以水及乙醇萃取，可得四種草藥之水及乙醇萃取物。本研究建立類巨噬細胞之U937細胞株，以cell counting kit-8方法進行萃取物之細胞毒性試驗，及促進U937細胞株吞噬標示綠色螢光蛋白之克雷白氏肺炎桿菌作用之篩選平台，將此平台進行四種台灣草藥之水及乙醇萃取物篩選。結果顯示九節茶葉部乙醇萃取物 (CGLE) 在100 μg/ml 濃度對 U937 細胞仍有最低毒性及最佳吞噬能力。因此，進行 CGL 之乙醇大量 (1.43 Kg) 萃取，並以 CH2Cl2 及 H2O 溶媒進行分配萃取，可得 CH2Cl2 層、乳化層及 H2O層等萃取物。將此三層萃取物進行細胞毒性試驗及促進 U937 細胞株吞噬作用，結果顯示CH2Cl2 層在50 μg/ml 濃度仍有99.8％之 U937 細胞存活率，於100 μg/ml 濃度則有最佳 (62.4％) 之促進 U937 吞噬能力。因此，進行CH2Cl2 層之矽膠管柱層析分離，以CH2Cl2：MeOH (60：1) 溶媒沖提，可得 CGLEC 1 ~ CGLEC 6 六個分層。再將此六個分層進行細胞毒性試驗及促進 U937細胞株吞噬作用，結果顯示 CGLEC 5於100 μg/ml對U937細胞株仍有97％之細胞存活率及73.5％之最佳吞噬效果。 CGLEC 5之有效成分有待進一步分離。 The purpose of this study is to set up a screening platform for immunity-enhancing function and isolate the effective constituents from Taiwanese herbal medicines (THMs) through this plateform. Four herbal medicines, including the leaves and stem of Chloranthus glaber (CGL, CGS), Daemonorops margaritae (DM), Eupatorium formosanum (EF) and Solanum nigrum (SN), were collected in Taiwan. They were respectively extracted with H2O and EtOH, and H2O and EtOH extracts were obtained. U937 cells, kind of macrophage-like cells, were used to do the cytotoxicity test by cell counting kit-8 method. The number of Klebsiella pneumoniae-gfp Phagocytosed by U937 cells was used as phagocytosis-enhancing assay by THMs. These two methods were applied to screen H2O and EtOH extracts of four THMs, respectively. The results showed that EtOH extracts of the leaves of CG (CGLE) at 100 μg/ml had the lowest cytotoxicity and was the most effective in enhancing phagocytosis of U937 cells. So a large amount (1.43 Kg) of CGL was extracted with EtOH and then partitioned and extracted with CH2Cl2 and H2O solvents. Three layers, namely, CH2Cl2 layer, emulsion layer and H2O layer extracts were respectively obtained. The three layers extracts were examined by cytotoxicity and phagocytosis tests. The results indicated that the CH2Cl2 layer extract at 50 μg/ml still had 99.8％ viability of U937 cells and at 100 μg/ml was the most effective (62.4％) in enhancing phagocytosis of U937 cells. The CH2Cl2 layer extract was subjected to chromatography on a silica gel column and eluted with CH2Cl2-MeOH (60：1) to give six fractions. The six fractions were again screened by cytotoxicity and phagocytosis tests. CGLEC 5 at 100 μg/ml still had 97％ viability of U937 cells and was the most effective (73.5％) in enhancing phagocytosis of U937 cells. The effective constituents to enhance immune system in CGLEC 5 remains to be isolated.
Relation:	校內校外均不公開
Appears in Collections:	[Dept. of Pharmacy] Dissertations and Theses

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