目的:研究Fas信号在同型半胱氨酸致人脐静脉内皮细胞凋亡过程中的作用。方法:以人脐静脉内皮细胞株为实验对象,用含不同浓度的同型半胱氨酸的DMEM培养液对其处理,MTT法测定细胞增殖能力;流式细胞术测定细胞凋亡水平;荧光定量PCR测定Fas基因、caspase-3基因及caspase-8基因的mRNA表达;Western blot测定Fas蛋白、caspase-3蛋白及caspase-8蛋白表达量。结果:同型半胱氨酸对人脐静脉内皮细胞的增殖能力有明显抑制作用,随着同型半胱氨酸浓度增高,实验结果显示细胞增殖能力逐渐降低,细胞凋亡率呈上升现象;与对照组相比,实验组Fas、caspase-3及caspase-8mRNA及蛋白表达明显上升。结论:同型半胱氨酸可能通过上调Fas、caspase-3及caspase-8的mRNA及蛋白的表达,促进人脐静脉内皮细胞的凋亡。
Objective:To study the role of Fas in human umbilical endothelial cell apoptosis induced by homocysteine(Hcy).Method:Human umbilical endothelial cells(HUVEC)were treated with different concentrations of Hcy,MTT was used to measure the proliferation of HUVEC.The flow cytometry assay was used to detect the level of cell apoptosis.The expressions of Fas,caspase-3and caspase-8were correspondingly analyzed by fluorescence quantitative PCR and Western blotting.Result:The proliferation of HUVEC was obviously inhibited by homocysteine.The rate of apoptosis in HUVEC cells increased.Compared to the control group,the expression of Fas and caspase-8and caspase-3mRNA and protein significantly increased.Conclusion:Hcy might induce apoptosis of HUVEC through up-regulating the Fas,caspase-3and caspase-8expression.