RIG-I配体poly(dAT:dAT)诱导Bewo细胞抑制乙型肝炎病毒复制的研究  

Study of RIG-I ligand poly(dAT:dAT)-induced inhibition hepatitis B virus replication in Bewo cells

作　　者：王芳 资捷 刘泽滨 方小龙 邱丽影 朱国胜 WANG Fang;ZI Jie;LIU Ze - bin;FANG Xiao - long;QIU Li - ying;ZHU Guo - sheng(Shenzhen Futian Women and Children Health Care Hospital, Shenzhen Guangdong 518026, P. R. China)

机构地区：[1]深圳市福田区妇幼保健院,广东深圳518026

出　　处：《中国计划生育和妇产科》2017年第11期58-62,共5页Chinese Journal of Family Planning & Gynecotokology

基　　金：深圳市科创委基金资助项目(编号:JCYJ20150402154553256)

摘　　要：目的探讨维甲酸诱导基因-I(retinoic acid inducible gene I,RIG-I)配体poly(d AT:d AT)抑制人滋养层细胞Bewo中乙型肝炎病毒(hepatitis B virus,HBV)复制的作用及机制,为防治HBV宫内感染提供依据。方法首先将2μg 1.3倍HBV全基因重组质粒pc DNA3.1(+)-HBV1.3转染Bewo细胞12 h后,以RIG-I配体poly(d AT:d AT)处理24 h。然后,以poly(d AT:d AT)处理Bewo细胞,观察干扰素-β(interferon-β,IFN-β)表达的动力学。采用微粒子酶免疫分析法和荧光定量聚合酶链反应法分别检测HBs Ag、HBe Ag和HBV DNA水平,并以酶联免疫吸附测定和逆转录PCR分别检测IFN-β水平,RIG-I、线粒体抗病毒信号蛋白(mitochondrial antiviral signaling protein,MAVS)、干扰调节因子3(interferon regulatory factor 3,IRF 3)和核因子-κB(NF-κB)的表达。结果 poly(d AT:d AT)可显著诱导Bewo细胞产生IFN-β(P<0.05),呈时间和剂量依赖性。与对照组比较,poly(d AT:d AT)处理组HBs Ag、HBe Ag和HBV DNA水平显著下降(P<0.05),可显著抑制Bewo细胞中HBV复制。且poly(d AT:d AT)可诱导HBV重组质粒转染的Bewo细胞表达RIG-I、MAVS、IRF 3和NF-κB mRNA。结论通过RIG-I/MAVS信号途径诱导Bewo细胞产生IFN-β,RIG-I配体poly(d AT:d AT)可显著抑制HBV复制。Objective To explore the effect and mechanism of retinoic acid inducible I （ RIG - I） ligand poly （ dAT： dAT） - mediated inhibition hepatitis B virus （HBV） replication in Bewo cells. Methods Firstly, 2μg 1.3 - fold HBV recombinant vector pcDNA3.1 （ ＋ ） - HBV1.3 were transfected into Bewo ceils. After 12 h the cells were treated with RIG - I hgand poly（dAT：dAT） for 24 h. Then Bewo ceils were exposed to poly （dAT：dAT） to observe the kinetics of interferon - β （IFN - β） expression in Bewo ceils. To detect HBsAg, HBeAg and HBV DNA level in the culture supernatant by microparticle enzyme immunoassay （MEIA） and fluorescence quantitative polymerase chain reaction （PCR）. And to assay IFN - β level and RIG - I, MAVS, IRF3, NF - κB reRAN expression by ELISA and RT - PCR, respectively. Results Compared with control, poly （ dAT ： dAT） could striking induce the production of IFN -β in Bewo cells（P 〈0. 05） ,in time - and dose -dependent manners. And it could significantly suppresse HBV replication in Bewo cells （ P 〈 0. 05 ）. Compared with control group, the mRNA levels of RIG - I, MAVS, IRF3 and NF - κB were significantly induced by poly（dAT：dAT） in the Bewo cells transfected with this recombinant vector. Conclusion RIG - I ligand poly （dAT：dAT） could significantly suppress HBV replication by inducing IFN -β production in Bewo ceils mainly via the RIG - I/MAVS signal pathway.

关 键 词：维甲酸诱导基因-I 线粒体抗病毒信号蛋白 乙型肝炎病毒 病毒复制 信号传导 

分 类 号：R512.62[医药卫生—内科学]