乙型肝炎病毒X蛋白通过激活Fas/Fas配体死亡受体途径促进肾小管上皮细胞凋亡  

Hepatitis B virus X protein induces apoptosis in renal tubular epithelial cells through upregulating death receptor pathway

作　　者：何平[1] 闵柳畅 张蓓茹[1] 刘大军[1] 杨旭[1] 周光宇[1] 边晓慧[1] 王艳秋[1] 李德天[1] 

机构地区：[1]中国医科大学附属盛京医院肾内科,辽宁沈阳110004

出　　处：《上海医学》2016年第1期16-19,共4页Shanghai Medical Journal

基　　金：辽宁省博士科研启动基金(201501005);辽宁省教育厅科研资助项目(L2013295)资助

摘　　要：目的探讨乙型肝炎病毒X蛋白(HBx)诱导大鼠肾小管上皮细胞凋亡及其可能的机制。方法以HBx的表达载体转染体外培养的大鼠肾小管上皮细胞(NRK-52E)建立HBx过表达模型。分别转染空质粒pc-DNA3.1(+)24h(空质粒转染组),以及转染pc-DNA3.1(+)HBx质粒24h(HBx24h组)、48h(HBx48h组)和72h(HBx72h组),并设对照组(未做转染)。应用流式细胞仪测定细胞凋亡率。分别采用Western印迹法检测Fas、Fas配体(FasL)、Bcl-2、Bax的蛋白质表达水平,分光光度测定法测定半胱氨酸天冬氨酸蛋白酶(Caspase)8活性。结果 HBx24h组、HBx48h组、HBx72h组的HBx、Fas、FasL、Bax的蛋白质相对表达量,以及Caspase8的相对活性逐渐增加,且组间差异均有统计学意义(P值分别<0.05、0.01);Bcl-2的蛋白质相对表达量逐渐降低,组间差异亦有统计学意义(P值分别<0.05、0.01)。对照组与空质粒转染组间细胞凋亡率的差异无统计学意义(P>0.05);HBx24h组、HBx48h组和HBx72h组的细胞凋亡率逐渐升高,组间差异有统计学意义(P值均<0.05),且均显著高于对照组和空质粒转染组(P值均<0.05)。结论 HBx基因通过Fas/FasL死亡受体途径促进NRK-52E细胞凋亡,Caspase8的活化可能参与其中。Objective To investigate the mechanism of hepatitis B virus X protein（HBx）-induced apoptosis in rat renal tubular epithelial cells（NRK-52E）.Methods An HBx expression vector（pc-DNA3.1／[＋／]-HBx）was used to transfect NRK-52 Ecells to establish an HBx overexpression model.A control group was not transfected and another control group was transfected with plasmid lacking the HBx-encoding insert.The rate of apoptosis was determined by flow cytometry.The expression of Fas,Fas ligand（FasL）,B cell lymphoma/lewkmia-2（Bcl-2）,and Bcl-2associated X protein（Bax）were determined by Western blotting,and the activity of Caspase8 was measured by spectrophotometry.Results Transfection of NRK-52 Ecells with pc-DNA3.1（＋）-HBx led to inhibition of proliferation and increased apoptosis of NRK-52 Ecells relative to the controls.Transfected cells had increased protein expression of Fas,FasL,and Bax and decreased protein expression of Bcl-2relative to the controls（P〈0.05 or 0.01）.In addition,transfected cells had increased Caspase8 activity relative to the controls.Conclusion Our results suggest that HBx can induce apoptosis in NRK-52 Ecells,at least in part through activation of the Fas/FasL pathway.The activation of Caspase8 appears to mediate the induction of apoptosis.

关 键 词：细胞凋亡 Fas FAS配体 乙型肝炎病毒X蛋白 肾小管上皮细胞 

分 类 号：R692[医药卫生—泌尿科学] R512.62[医药卫生—外科学]