丙肝病毒核心蛋白抑制乙肝病毒复制的研究  

Study on inhibition of HCV core protein to the replication of HBV

作　　者：应桑雨[1] 高国生[1] 徐晓珍[1] 陆传统 

机构地区：[1]宁波市第二医院,浙江宁波315010 [2]宁波市卫生信息中心,浙江宁波315010

出　　处：《中国卫生检验杂志》2016年第6期772-774,共3页Chinese Journal of Health Laboratory Technology

基　　金：宁波市自然科学基金(2012A610248;2012A610183)

摘　　要：目的探讨丙肝病毒(HCV)核心蛋白(Core)对乙肝病毒(HBV)复制的影响,并探讨其调节机制。方法将Core蛋白真核表达载体Flag2B-Core、HBV感染性克隆p HBV1.3及其启动子p HBV-Luc分别转染Hep G2和Hep G2.2.15细胞;酶联免疫吸附试验(ELISA)检测细胞上清液中HBs Ag和HBe Ag含量;荧光定量PCR法检测细胞闭合环状DNA(ccc DNA);Luminometer荧光检测仪检测HBV启动子活性。结果随着HCV Core蛋白浓度的不断增加,Hep G2.2.15细胞上清中的HBs Ag和HBe Ag表达逐渐降低,0.0μg组、0.5μg组均显著高于1.0μg组、2.0μg组(P<0.05);Hep G2.2.15细胞内HBV ccc DNA含量亦逐渐降低,并呈剂量依赖效应;Hep G2细胞荧光素酶的活性不断降低,0.0μg组、0.5μg组均显著高于1.0μg组、2.0μg组(P<0.05),Core蛋白下调HBV启动子活性呈剂量依赖效应,Spearman相关分析显示两者相关性良好(P<0.01)。结论 HCV Core蛋白在细胞水平抑制HBV的复制。Objective To explore the inhibition effect of hepatitis C virus（ HCV） core protein on the replication of Hepatitis B virus（ HBV）,and to explore its regulatory mechanism. Methods Hep G2 and Hep G2. 2. 15 cells were infected with plasmid carrying Core,Flag2B- Core,HBV infectious clone p HBV1. 3 and its promoter p HBV- Luc,respectively; HBs Ag and HBe Ag in the supernatant were measured by enzyme linked immunosorbent assay（ ELISA）; closed circular DNA（ ccc DNA） levels were measured by real- time PCR,and HBV promoter activity was measured by luminometer fluorescence detector. Results With the increasing of HCV Core protein concentration,expression of Hbs Ag and HBe Ag in Hep G2. 2. 15 supernatant were downregulated,and the groups of 0. 0 μg and 0. 5 μg were much higher than the groups of 1. 0 μg and 2. 0 μg（ P〈0. 05）; HBV ccc DNA in Hep G2. 2. 15 was also downregulated and had a dose- dependent effect; the activity of Hep G2 luciferase decreased,and the groups of 0. 0 μg and 0. 5 μg were much higher than the groups of 1. 0 μg and 2. 0 μg（ P〈0. 05）,hence,HBV promoter activity was downregulated by Core protein in a dose- dependent manner,indicating that there was a good correlation between them by Spearman correlation analysis（ P〈0. 01）. Conclusion HCV Core protein inhibits the replication of HBV at the cell level.

关 键 词：乙肝病毒 核心蛋白 酶联免疫吸附 复制 

分 类 号：R373.21[医药卫生—病原生物学]