凋亡相关蛋白在汉坦病毒诱导HEK-293细胞凋亡过程中的表达及意义

目的 探讨Bcl-2、Bax及Caspase-3在汉坦病毒诱导人胚肾细胞(HEK-293)凋亡过程中的变化,为研究汉坦病毒诱导人胚肾细胞损伤的机制提供参考.方法 体外培养HEK-293细胞,分为正常对照组和汉坦病毒处理的感染组,采用间接免疫荧光法检测HEK-293内汉坦病毒抗原；用Westen blot方法检测Bcl-2、Bax及Caspase-3蛋白表达水平.结果 汉坦病毒感染HEK-293细胞1d后即开始出现荧光阳性细胞,随着时间延长,荧光强度逐渐增强,细胞胞浆内出现大量片状或颗粒性的黄绿色荧光；Western blot结果显示,与对照组比较,汉坦病毒感染1d后Bcl-2、Bax蛋白及Caspase-3酶原活化片段表达量未见明显变化(P＞0.05),感染3d和5d后Bcl-2蛋白表达降低,Bax蛋白表达上调,Caspase-3酶原活化片段表达升高(P＜0.05).结论 汉坦病毒可感染HEK-293细胞并在其体内增殖,其损伤机制可能与Bcl-2表达减少和Bax蛋白表达上调,通过线粒体途径诱导HEK-293细胞发生凋亡有关.

Expression and significance of relative factors in HEK-293 apoptosis induced by hantavirus

Objective To provide reference to possible mechanism of injury in human embryonic kidney 293 cell(HEK-293),which was induced by hantavirus by means of investigating the variation of Bcl-2、Bax and Caspase-3.Methods HEK-293 in vitro culture was divided into normal control group and infection group processed by hantavirus.Hantavirus antigen in HEK-293 cells were detected by indirectimmunofluorescent assay and the expression of Bcl-2、Bax and Caspase-3 were assessed by Western blot.Results Fluorescence-positive cells emerged one day after HEK-293 cells were infected with hantavirus,and with time going on,fluorescence intensity increased gradually,as well as a large amount of foliated or granular greenyellow fluorescence in intracytoplasm.As Western blot results showed,in contrast to control group,the expression of Bcl-2、Bax and Caspase-3 did not present marked change 1 day postinfection(P>0.05),and the expression of Bcl-2 decreased with that of Bax and Caspase-3 activation increasing 3 or 5 days postinfection(P<0.05).Conclusion The injury mechanism produced by hantavirus which infected and proliferated in HEK-293 cell could be relative to the decrease of Bcl-2 expression and the up-regulation of Bax protein,as well as apoptosis of HEK-293 cells induced by mitochondrial-mediated manner.