二苯乙烯苷对氧化应激诱导的内皮细胞凋亡的影响

项目名称： 二苯乙烯苷对氧化应激诱导的内皮细胞凋亡的影响

项目编号： No.30800474

项目类型： 青年科学基金项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 龙石银

作者单位： 南华大学

项目金额： 19万元

中文摘要： 血管内皮细胞受损是触发动脉粥样硬化（As）的始动因素之一，抗内皮损伤、保护内皮功能是防治As的重要靶点。二苯乙烯苷（TSG）是何首乌主要的药效成分，具有抗氧化、降血脂和防治动脉粥样硬化作用，但机制未明。本项目以过氧化氢诱导人脐静脉内皮细胞损伤，采用MTT法、细胞形态学观察、流式细胞仪分析等分别筛选了合适的过氧化氢浓度造模人脐静脉内皮细胞氧化损伤和细胞凋亡、以及TSG的最佳作用浓度，并运用RT-PCR、Western blot、ELISA等方法检测了TSG对氧化损伤内皮细胞炎症应答相关基因（NF-κ#12289;Iκ#12289;TNF-α12289;IL-6），粘附分子（P-selectin、E-selectin、ICAM-1、VCAM-1和MCP-1）以及凋亡相关基因（Caspase-3、bcl-2、bax、PARP、p53）的表达，初步探讨了TSG抗氧化、抗内皮细胞凋亡的分子信号机制。结果显示TSG能调节氧化诱导损伤的内皮细胞中上述炎症应答相关基因、粘附分子及凋亡分子的表达，具有抗氧化、抗凋亡，保护内皮细胞的作用，本研究对确定二苯乙烯苷的分子作用靶点及开发应用具有重要的意义。

中文关键词： 二苯乙烯苷；人脐静脉内皮细胞；细胞凋亡；氧化应激

英文摘要： Vascular endothelial cell damage was one of the initiating factors of trigger atherosclerosis (As). Anti-endothelial injury, protecting of endothelial function were the important target for prevention and treatment of atherosclerosis. 2,3,5,4′tetrahydroxystilbene-2-O-β-glucoside (TSG) was a pharmacological compounds in Polygonum multiflorum, and had the roles of antioxidant, anti-atherosclerosis, and could reduce the lever of lipid, but the mechanism was unknown, so the aim of this study was to explore the effects of TSG and its molecular mechanisms on oxidative stress human umbilical vein endothelial cells (HUVEC) induced by hydrogen peroxide. The concentrations of hydrogen peroxide and TSG on HUVEC were established via MTT, cell morphological, Hoechst33258 staining, and Flow Cytometry. The expressions of NF-κ Iκ TNF-alpha, IL-6, P-selectin, E-selectin, ICAM-1, VCAM-1, MCP-1, Caspase-3, bcl-2, bax, PARP and p53 were detected by RT-PCR, Western blot and ELISA respectively. The results showed that the TSG could regulate the expression of above mentioned inflammatory response related genes, adhesion molecules and related apoptotic molecules in oxidation-induced damage of endothelial cells. TSG had the role of antioxidant, anti-apoptotic protecting effects in endothelial cells, and the molecular mechanism might be related with NF-κ Iκsignal pathway and apoptosis signal pathway. This study might contribute to determine the exact molecular targets, and benefit to the development and application of TSG.

英文关键词： 2; 3; 5; 4′Tetrahydroxystilbene-2- O-β- glucoside(TSG); Human vein endothelial cell(HUVEC); Apoptosis; Oxidative stress