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研究生(外文):Thu-Ching Hung
論文名稱(外文):Effect of Arecoline on Oxidative stress in Human Umbilical Vein Endothelial Cells
指導教授(外文):Kee-Lung Chang
外文關鍵詞:ArecolineHuman Umbilical Vein Endothelial CellsHO-1
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本研究發現人類血管內皮細胞經檳榔素刺激後,細胞週期會停滯在G2/M期,並抑制DNA合成。檳榔素會促進人類臍靜脈內皮細胞產生大量ROS,並促進細胞黏著分子intercellular cell adhesion molecule-1 (ICAM-1)及vascular cell adhesion molecule-1 (VCAM-1)大量表現。由於細胞內ROS的產生,造成細胞內抗氧化酵素及成分含量減少:包括superoxide dismutase (SOD)、catalase以及glutathione (GSH)。在加入檳榔素處理的細胞中再加入抗氧化劑GSH、N-acetyl-L-cysteine (NAC)可以減少ROS的產生與ICAM-1及VCAM-1的表現,而加入buthionine sulfoximine (BSO) 則會有加重的情形。也發現檳榔素會促使氧化壓力蛋白HO-1大量表現,利用HO-1 抑制劑zinc protoporphyrin-Ⅸ (ZnPP) 則會增加細胞的氧化傷害及ICAM-1與VCAM-1 的表現增加。實驗中分別利用ERK、JNK、p38 MAPK以及tyrosine kinase pathway的抑制劑加以作用,發現檳榔素主要是藉由JNK、p38 MAPK以及tyrosine kinase pathway調控細胞內HO-1的表現,而非ERK pathway。並利用細胞轉殖的方式,使人類臍靜脈內皮細胞大量表現HO-1蛋白,實驗結果得知,HO-1的大量表現可抑制檳榔素對於血管內皮細胞的毒性作用。

Arecoline, the most abundant areca alkaloid, has been reported to stimulate reactive oxygen species (ROS) production in several cells. Overpeoduction of ROS has been implicated to atherogenesis. Hemeoxygenase-1 (HO-1) has cytoprotective activities in vascular tissues. This study investigated the effect of arecoline on the pathogenesis of atherosclerosis and explored the role of HO-1 in this process. Human Umbilical Vein Endothelial Cells (HUVECs) were treated with arecoline, followed by analysis of ROS, adhesion molecules, and HO-1 expressions, meanwhile, the potential signaling pathways were determined. After arecoline treatment, ROS production was stimulated, adhesion molecules ICAM and VCAM expressed coincidently. Glutathione pretreatment could totally inhibit arecoline-stimulated ROS production and VCAM expression but not ICAM expression. Arecoline simultaneously induced HO-1 expression and which was partially relative to ROS stimulation. Inhibition of c-jun N-terminal kinase (JNK) by SP600125, p38 by SB 203580 or tyrosine kinase by genistein reduced arecoline-induced HO-1 expression. In contrast, inactivation of ERK (extracellular signal-related MAP kinase) by PD98059, had no effect. Transfection of GFP/HO-1 gene to HUVEC with 5-folds HO-1 activities could efficiently but not completely attenuate the decrease in cell viability by arecoline. This study demonstrates in HUVEC arecoline stimulated ROS production associated with ICAM and VCAM expressions. Under this condition, HO-1 expression was up-regulated through ROS, tyrosine kinase and MAPK (JNK and p38) signaling pathways.

目 錄

壹、中文摘要----------------------------------------------- 1
貳、英文摘要----------------------------------------------- 3
參、前言----------------------------------------------------- 4
血管內皮細胞----------------------------------------- 4
動脈粥狀硬化----------------------------------------- 9
檳榔---------------------------------------------------- 19
自由基------------------------------------------------- 23
血紅素氧化酶---------------------------------------- 26
肆、研究動機與目的------------------------------------- 31
伍、實驗材料---------------------------------------------- 32
陸、實驗方法---------------------------------------------- 39
柒、實驗結果---------------------------------------------- 54
捌、討論---------------------------------------------------- 63
玖、參考文獻---------------------------------------------- 73
拾、圖表--------------------------------------------------- 102
拾壹、附錄------------------------------------------------ 135

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