腺病毒介导AMP激活的蛋白激酶的过表达诱导肝星状细胞株LX2凋亡  

Adenovirus-mediated overexpression of AMP-activated protein kinase induces LX2 cell apoptosis

作　　者：张婧[1] 谭丽[1] 林骏[1] 徐爱民[2] 罗深秋[1] 

机构地区：[1]南方医科大学细胞生物教研室,广东广州510515 [2]香港大学医学院

出　　处：《南方医科大学学报》2007年第6期821-823,共3页Journal of Southern Medical University

基　　金：广东省自然科学基金(05102580)~~

摘　　要：目的探讨在肝星状细胞株LX2中,用腺病毒介导AMP激活的蛋白激酶(AMPK)过表达对细胞凋亡的影响。方法病毒感染细胞后,用免疫印记的方法检测外源基因的表达;流式细胞仪分析细胞的凋亡;琼脂糖凝胶电泳检测DNA梯状条带,免疫印迹检测caspase-3以及Bcl-2、Bax表达的变化。结果免疫印迹结果显示表达组成性激活的AMPK的腺病毒感染LX2细胞72h,AMPK过表达。过表达AMPK的LX2细胞用流式细胞仪检测到有凋亡的亚二倍体峰出现;DNA断裂出现梯状条带,casepase-3被激活,Bcl-2在LX2中是低表达的,而Bax表达水平升高。结论腺病毒介导AMPK的过表达可以诱导LX2细胞发生凋亡,其可能的机制之一是上调促凋亡基因bax的表达。Objective To observe the effect of adenovirus-mediated overexpression of AMP-activated protein kinase （AMPK） on apoptosis of hepatic stellate cell line LX2. Methods After adenovirus infection of the LX2 cells, the exogenous gene expression was detected by Western blotting, and cell apoptosis evaluated by flow cytometry with PI staining. Agarose gel electrophoresis was performed to detect the DNA ladder, and the expressions of caspase-3, Bcl-2 and Bax are detected by Western blotting. Result With AMPK overexpression, apoptotic peak and DNA ladder of the infected cells appeared, and pro-caspase-3 was activated to transform into caspase-3 accompanied by up-rugulated Bax expression, whereas Bcl-2 expression was hardly detectable by Western blotting. Conclusion Overexpression of AMPK mediated by the adenovirus can induce LX2 cell apoptosis, possible as a result of up-regulated Bax expression with low Bcl-2 expression.

关 键 词：肝纤维化 肝星状细胞 腺病毒 凋亡 

分 类 号：Q343[生物学—遗传学]