【摘要】 目的:探讨肝细胞生长因子抑制糖基化终产物诱导内皮细胞凋亡的作用及其相关分子机制。方法:体外培养人脐静脉内皮细胞,分为实验对照组、糖基化终产物及糖基化终产物+肝细胞生长因子组,采用MTT法测定各组血管内皮细胞生长抑制率;瑞氏-吉姆萨染色观察细胞形态学变化、流式细胞术测定细胞凋亡率;蛋白免疫印迹法分析各组凋亡基因Bax、Bc l-2蛋白的表达及酶联反应法测定caspase-3活性。结果:糖基化终产物诱导培养的内皮细胞出现明显的凋亡形态学改变,内皮细胞凋亡率与糖基化终产物的浓度和作用时间呈依赖关系,肝细胞生长因子干预后可显著降低不同时间的内皮细胞凋亡率;肝细胞生长因子作用内皮细胞抗凋亡基因Bc l-2表达明显升高,而促凋亡基因Bax表达无明显变化;caspase-3活性显著降低。结论:肝细胞生长因子抑制糖基化终产物诱导内皮细胞凋亡,其作用机制可能是上调抗凋亡基因Bc l-2水平、抑制caspase-3的激活。更多还原

【Abstract】 Objective: To investigate the inhibitory effect of hepatocyte growth factor(HGF) on vascular endothelial cell apoptosis induced by advanced glycosylation end products(AGEs) and the possible mechanism.Methods: Human umbilical vein endothelial cells(HUVECs)were cultured in vitro and treated with different concentrations of AGEs and HGF.The growth inhibition rates of the cells in each group at different time points (12,24,48,and 72 hours after intervention) were determined by methyl thiazolyl tetrazolium(MTT) assay.The cell apoptosis was detected by flow cytometry,and the cell morphology was observed by Wright’s-Giemsa staining.The expressions of apoptosis-associated genes Bax and Bcl-2 were detected by Western blotting,and the activity of caspase-3 was measured by enzyme-linked immunosorbent assay.Results: Morphological observation indicated that AGEs induced characteristic apoptotic changes in HUVECs in a dose-and time-dependent manner.HGF significantly inhibited the apoptosis of HUVECs induced by AGEs.HGF significantly increased the expression of Bcl-2 and decreased the activity of caspase-3,without affecting Bax expression.Conclusion: AGEs can induce the apoptosis of vascular endothelial cells in vitro.HGF effectively inhibits AGEs-induced apoptosis by upregulating the expression of Bcl-2 and inhibiting the activation of caspase-3.更多还原