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研究生:徐詩雯
研究生(外文):Shih-Wen Hsu
論文名稱:建立血管發炎的離體模型並探討生藥粉防己對內皮細胞ICAM-1表現及嗜中性白血球的黏附作用
論文名稱(外文):The establishment of an in vitro vascular inflammation model and its application for studying the effect of herbal medicine Stephania tetrandra S. Moore in ICAM-1expression of endothelial cells and the neutrophil-endothelial cell adhesion
指導教授:廖志飛廖志飛引用關係汪貴珍汪貴珍引用關係
指導教授(外文):Jyh-Fei LiaoGuei-Jane Wang
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:藥理學研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:中文
論文頁數:72
中文關鍵詞:細胞黏附分子嗜中性白血球粉防己粉防己鹼
外文關鍵詞:cellular adhesion moleculesneutrophilStephania tetrandra S. Mooretetrandrine
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抑制細胞黏附分子的表現,對於控制各種發炎性疾病非常重要。本研究建立一個體外發炎模式,探討粉防己部分純化抽提物 (partial purified extract of Stephania tetrandra S. Moore)及其生物鹼粉防己鹼 (tetrandrine)的抗發炎作用。
以西式墨點法來觀察TNF-a (0.1 ng/ml)引起人類臍靜脈內皮細胞ICAM-1之表現。實驗結果顯示,內皮細胞處理粉防己部分純化抽提物 (10 mg/ml)或粉防己鹼 (130-260 ng/ml)可抑制TNF-a活化ICAM-1的表現,抑制率分別為48.8 %及83.9%-88.9%。以螢光顯微鏡與微盤螢光光度儀兩種方式,觀察PKH2螢光劑標誌之的嗜中性白血球與內皮細胞間的黏附作用。粉防己部分純化抽提物及粉防己鹼均可抑制細胞間的黏附作用,但此抑制作用也許不經內皮細胞產生。綜合以上結果,粉防己部分純化抽提物及粉防己鹼可以抑制內皮細胞表面ICAM-1表現,以及嗜中性白血球與內皮細胞在共同培養模式中的黏附作用。但是內皮細胞ICAM-1的表現並不能完全決定其對嗜中性白血球的黏附作用,防己部分純化抽提物及粉防己鹼所表現的抑制作用可能是由於抑制了白血球的活化。
Inhibition of the expression of cell adhesion molecules has been shown to be important in controlling various inflammatory diseases. To study the anti-inflammatory effects of partial purified extract of Stephania tetrandra S. Moore and tetrandrine, an in vitro vascular inflammation model was established.
TNF-a (0.1 ng/ml)-stimulated ICAM-1 expression in human umbilical vein endothelial cells (HUVECs) was analyzed by Western blotting. Both the partial purified extract of Stephania tetrandra (10 mg/ml) and tetrandrine (130-260 ng/ml) could inhibit TNF-a-stimulated ICAM-1 expression in HUVECs by 48.8% and 83.9%-88.9%, respectively. The neutrophil-endothelial cell adhesion measured by fluorescent PKH2 labeling of neutrophil using a fluorescence microscope and microplate reader. Both the partial purified extract of Stephania tetrandra and tetrandrine inhibited neutrophils adhere to HUVECs induced by TNF-a. However, the inhibitory effects on neutrophil-HUVECs adhesion of the partial purified extract of Stephania tetrandra and tetrandrine were not related to their inhibition on ICAM-1 expression of HUVECs. Taken together, the present results demonstrated that the partial purified extract of Stephania tetrandra and its active alkaloid tetrandrine could inhibit TNF-a-induced ICAM-1 expression of HUVECs and neutrophil-HUVEC adhesion in coculture system. The inhibition of the partial purified extract of Stephania tetrandra and tetrandrine on neutrophil-HUVEC adhesion may be dependent on neutrophil activation by TNF-a.
謝誌
目錄
縮寫表…………………………………………………………………1
中文摘要………………………………………….………….………2
英文摘要………………………………………………………………3
壹、緒論………………………………………….………………….5
一、 發炎的防禦機制…………………………………….………..5
二、 黏附分子………………………………………..……...……6
三、 嗜中性白血球的外滲作用……………………………..……10
四、 生藥粉防己及其生物鹼粉防己鹼的生物活性……....……11
貳、研究動機….……………………………….……………..….13
參、實驗材料與方法…………………………………….………..15
肆、實驗結果……………………………….………………...….28
伍、討論………………………………………..…………...……37
參考文獻…………………………………………………………….42
圖…………………………………………..……………………….47
附註……………/…………………………………………………..62
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中藥大辭典 (1977) 上海科技出版社
蘇純閨 編譯,藥理學 (1998) 台灣:合記圖書出版社
鄭永銘 編譯,基礎病理學(1999)台灣:合記圖書出版社
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