【作者】 贾德安；

【导师】 朱鹏立；

【作者基本信息】 福建医科大学 ， 老年医学， 2006， 硕士

【摘要】 背景：红葡萄酒的心血管保护作用可归功于其中的多酚类物质，尤其是白藜芦醇，而白藜芦醇的抗动脉粥样硬化作用源于其强大的抗炎作用。由于动脉粥样硬化是一种以内皮功能不全为突出表现的炎症反应过程，因此研究白藜芦醇在内皮细胞炎症反应中的作用对于动脉粥样硬化的防治具有重大意义。CD40途径参与了动脉粥样硬化中主要细胞成分的炎症反应调节，本文初步探讨白藜芦醇是否能够通过影响CD40途径发挥心血管保护作用。 目的：本实验利用肿瘤坏死因子(TNF)-α、sCD40L活化体外培养的人脐静脉内皮细胞(HUVEC)，目的研究白藜芦醇对活化的HUVEC炎症基因表达及CD40途径的影响。 方法：将培养的HUVEC随机分组，分别为对照组、TNF-α(10μg／L)组、sCD40L(10μg／ml)组、不同浓度白藜芦醇+TNF-α组和白藜芦醇+sCD40L组。药物处理组实验顺序为白藜芦醇预孵育后加入TNF-α或sCD40L刺激，显微镜下观察细胞大体形态的改变并以流式细胞术检测HUVEC表面CD40、E-selectin分子的表达；半定量RT-PCR方法检测CD40、MMP-2、MMP-9、E-selectin、MCP-1基因的转录水平。 结果：HUVEC经TNF-α、sCD40L刺激24h后，可见细胞坏死脱落，残余细胞形态肿胀、排列稀疏，而经白藜芦醇预孵育的HUVEC只有少量细胞坏死脱落，残余细胞形态良好，排列规整。可见白藜芦醇对TNF-α、sCD40L损伤的HUVEC具有保护作用。 经过TNF-α或sCD40L刺激后，流式细胞术检测HUVEC表达CD40和E-selectin的阳性细胞百分比及平均荧光强度较对照组HUVEC明显升高(P＜0.01)；白藜芦醇(10μmol／L)预孵育的HUVEC，在经过TNF-α或sCD40L刺激后，其表面CD40及E-selectin分子的表达明显少于单用TNF-α或sCD40L刺激时的表达(P＜0.05)。 TNF-α刺激后可显著增加HUVEC的CD40(P＜0.05)、MMP-9(P＜0.05)、E-selectin(P＜0.01)、MCP-1(P＜0.01)基因转录水平，白藜芦醇在0.1μmol／L～10μmol／L范围内可呈浓度依赖性的抑制CD40、MMP-9、E-selectin、MCP-1基因转录水平更多还原

【Abstract】 BACKGROUND: The cardioprotective effect of red wine owes to its polyphenols,especially resveratrol. And the anti-atherosclerosis effect of resveratrol derives fromits powerful anti-inflammatory effect. Atherosclerosis is an inflammatory processcharacteristic of endothelial dysfunction, so it is of significant importance toinvestigate the role of resveratrol in inflammatory endothelial cells. CD40 pathwayparticipates in the regulation of inflammatory reaction of atherosclerosis, so it is alsoessential to investigate whether resveratrol can protect cardiovascular system throughCD40 pathway.OBJECTIVE: In this study, the effect of resveratrol on inflammation geneexpression and CD40 pathway of HUVEC stimulated by TNF-a or sCD40L wasobserved.METHODS: Divide the cultivated HUVEC randomly into groups as the control,TNF-α(10μg/L), sCD40L(10μg/ml), resveratrol of different concertrations +TNF-αand resveratrol +sCD40L(10μg/ml). The empirical sequence of medication group isTNF-a or sCD40L followed by the preincubation of resveratrol. Observe the changeof cells in morphology with microscope and detect the expression of CD40 andE-selectin molecule on the surface of HUVEC with flow cytometric analysis, anddetect the transcriptional level of CD40, MMP-2, MMP-9, E-selectin, MCP-1 genewith RT-PCR.RESULTS: Observation of cell morphology reveals that after the stimulation ofTNF-a or sCD40L, HUVEC necrosis and defluxio, remnant cells present a swellingmorphous and array sparsenessly. After the preincubation of resveratrol, only a fewcells necrosis and defluxio and the remnant cells present a fairly morphous. Itindicates that resveratrol has a protective effect on the TNF-a or sCD40L injuredHUVEC.After the stimulation of TNF-a or sCD40L, the positive cell percentage and mean更多还原