痉挛性截瘫蛋白21对乙型肝炎病毒复制的影响及其机制  

Study of the effect and mechanism of spastic paraplegia 21 protein on the repfication of hepatitis B virus

作　　者：高国生[1] 翁彭剑[1] 李永燕[1] 丁世雄[1] 

机构地区：[1]浙江省宁波市第二医院宁波市传染病医院,315010

出　　处：《中华肝脏病杂志》2011年第10期747-750,共4页Chinese Journal of Hepatology

基　　金：宁波市自然科学基金（2010A610056）

摘　　要：目的 探讨人痉挛性截瘫蛋白21（SPG21）对乙型肝炎病毒（HBV）复制的影响，并探讨其调节机制。方法将HBV感染性克隆pHBV1．3及其启动子pHBV—Luc分别转染HepG2细胞，加入不同浓度的SPG21蛋白，采用酶联免疫吸附法检测细胞上清液中HBsAg和HBeAg含量；RT—PCR和Westernblot法检测IlepG2细胞内HBV核心蛋白mRNA和蛋白表达；荧光定量PCR法检测细胞闭合环状DNA（cccDNA）的水平；采用Luminometer荧光检测仪分析HBV启动子活性的变化。实验数据用均数±标准差（x±s）表示，组间比较采用t检验。结果SPG21能够上调细胞上清液中HBsAg和HBeAg的含量，同时能够促进细胞内HBV核心蛋白的表达和HBVcccDNA的产生，并上调HBV启动子活性，其调节作用呈剂量依赖效应；与对照组比较，加入50μg／ml，100μg／ml，200μg／mlSPG21蛋白48h后，HBV启动子上调倍数分别为1．63、3．09和4．66（P〈0．05）。结论SPG21能够促进HBV在HepG2细胞中的复制。Objective To study the effect of human spastic paraplegia 21 protein （SPG21） on the replication of hepatitis B viru（HBV） and its regulatory mechanism. Methods HBV infectious clone pHBV1.3 and its promoter pHBV-Luc were transfected respectively into HepG2 cells with SPG21 of different concentrations, HBsAg and HBeAg in the supernatants were measured by enzyme linked immunosorbent assay （ELISA）, expression of HBV core mRNA and protein were detected by RT-PCR and western blot, covalently closed circular DNA（ccc DNA） levels were measured by real-time PCR, and HBV promoter activity was measured by luminometer fluorescence detector. Results Expression of HBsAg, HBeAg, HBV core protein and cccDNA were upregulated by SPG21 as well as HBV promoter activity in a dose-dependent approach. The activity of HBV promoter increased to 1.63, 3.09 and 4.66 times in HepG2 cells treated with 50μg/ml, 100μg/ml and 200μg/ml SPG21 respectively during 48 hour-treated （P〈 0.05）, as compared to the control group. Conclusions SPG21 can enhance the replication of HBV in HepG2 cells.

关 键 词：肝炎病毒 乙型 重组人痉挛性截瘫蛋白21 复制 

分 类 号：R51[医药卫生—内科学]