表没食子儿茶素没食子酸酯对晚期糖基化终产物诱导HUVEC凋亡的影响及可能的作用机制
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辽宁百千万人才工程入选资助项目(2012921033);辽宁省教育厅项目(L2011133);辽宁省自然科学基金(联合基金)(2013021022)


Effect of Epigallocatechin-3-gallate on Advanced Glycosylation End Product-induced Apoptosis in Human Umbilical Vein Endothelial Cell and the Possible Mechanism
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    目的 探讨表没食子儿茶素没食子酸酯(EGCG)对晚期糖基化终产物(AGE)诱导的人脐静脉内皮细胞(HUVEC)凋亡的影响及可能的作用机制。方法 用200 mg/L AGE培养HUVEC 24 h;以50、100 μmol/L EGCG预处理8 h后再用200 mg/L AGE作用于HUVEC;对照组用200 mg/L牛血清白蛋白作用于HUVEC 24 h。噻唑蓝比色法测定细胞活性,Hoechst-33258染色观察HUVEC形态变化,Annexin V-FITC/PI双标记法流式细胞仪检测细胞凋亡,活性氧检测以反映HUVEC内氧化应激水平。结果 与对照组相比,AGE组HUVEC发生染色质固缩、核碎裂等细胞凋亡的形态学改变,细胞活性降低(P<0.01),细胞凋亡率升高(P<0.01),细胞内活性氧水平增加(P<0.01)。EGCG预处理后能够使细胞凋亡的形态学改变明显减轻,细胞活性升高(P<0.01),细胞凋亡率减低(P<0.01),活性氧水平降低(P<0.01);EGCG作用呈浓度依赖性。结论 EGCG呈浓度依赖性抑制AGE诱导的HUVEC凋亡,这一作用可能通过抑制HUVEC内氧化应激反应而实现。

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    Aim To investigate the effect of epigallocatechin-3-gallate (EGCG) on advanced glycosylation end product (AGE) induced apoptosis in human umbilical vein endothelial cell (HUVEC) and the possible mechanism.Methods HUVEC was incubated with 200 mg/L AGE for 24 h in the presence or absence of different concentrations of EGCG (50,100 μmol/L) for further 8 h. The same concentration and action time of unmodified bovine serum albumin (BSA) was put into use as control group. Cell viability was evaluated by methyl thiazolyl tetrazolium (MTT) assay. Morphological change in HUVEC was observed by Hoechst-33258 staining,and cell apoptosis was analysed by Annexin V-FITC/PI double staining assay. Oxidative stress in HUVEC was evaluated by reactive oxygen species (ROS).Results Compared with BSA,AGE induced obvious morphological changes of cell apoptosis,such as chromatin pyknosis,karyorrhexis,reduced cell viability (P<0.01),increased apoptosis rate (P<0.01),enhanced oxidative stress reaction (P<0.01). Pretreatment with EGCG (50,100 μmol/L) in a concentration-dependent manner increased cell viability (P<0.01),relieved the morphological changes of cell apoptosis,decreased AGE-induced apoptosis and ROS production (P<0.01).Conclusion Our findings indicated that EGCG decreased AGE-induced apoptosis in HUVEC via the inhibition of oxidatie stress reaction.

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刘婷婷,孟 馨,金智一,贾 楠.表没食子儿茶素没食子酸酯对晚期糖基化终产物诱导HUVEC凋亡的影响及可能的作用机制[J].中国动脉硬化杂志,2015,23(05):480~484.

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  • 收稿日期:2014-05-23
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