Toll样受体激动剂抑制土拨鼠肝炎病毒作用研究  

Study of toll-like receptor (TLR) agonists on the inhibition of woodchuck hepatitis virus

作　　者：孟忠吉[1] 张永红[2] 李文兵[3] 汤守兵[1] 李东[1] 柯昌征[1] 陈悦[1] 

机构地区：[1]湖北医药学院附属太和医院感染科,十堰442000 [2]湖北医药学院附属太和医院中西医结合科,十堰442000 [3]湖北医药学院附属太和医院检验科,十堰442000

出　　处：《肝脏》2013年第6期378-381,共4页Chinese Hepatology

基　　金：湖北省科技厅自然科学基金重点项目(2010CDZ036);湖北医药学院基金项目(2009QD138)

摘　　要：目的研究Toll样受体(TLR)激动剂对土拨鼠肝炎病毒(WHV)复制的抑制作用。方法从WHV慢性感染的土拨鼠肝脏分离原代肝细胞,转染Poly I:C、CpG或直接用LPS刺激,Southern印迹检测WHV复制中间体,病毒保护试验检测细胞培养上清液中I型干扰素的水平;通过尾静脉注射Poly I:C、LPS、CpG到WHV转基因小鼠体内,提取肝脏DNA和RNA,实时PCR检测MxA和OAS1的mRNA水平。结果 Poly I:C、LPS和CpG在原代土拨鼠肝细胞和WHV转基因小鼠显著抑制病毒复制;Poly I:C、LPS和CpG处理后的原代肝细胞分泌高水平的I型干扰素,肝脏的MxA和OAS1 mRNA上调表达。结论TLR通路的活化通过诱导I型干扰素而发挥抗病毒作用,在抗HBV治疗中可能发挥重要作用。Objective To study the Toll-like receptor （TLR） agonists on the inhibition of woodchuck hepatitis virus （WHV）. Methods Primary woodchuck hepatocytes （PWHs） were isolated from woodchucks infected with WHV.. The hepatocytes were transfected with agonists of TLR-3 （Poly It C）, -9 （CpG） or directly stimulated with TLR-4 agonist （LPS）. WHV transgenic （Tg） mice were intravenously injected with Poly I： C, LPS and CpG from tail vein. WHV replication was detected using southern blot, and type I interferon （IFN） level in cell culture supernatant was detected using virus protection assay. The levels of mouse MxA and OAS1 mRNA were determined by real-time RT- PCR. Results WHV replication was inhibited significantly by Poly It C, LPS, and CpG treatment both in PWHs and in WHV Tg mice. Induced by Poly I： C, LPS, and CpG, high-level of type I IFN in the supernatants of PWHs was detected, and levels of MxA and OASI mRNA in the liver from WHV Tg mice were significantly up-regulated. Conclusion TLR agnoists can inhibit WHV replication through type I IFN induction, which might play an important role in anti-HBV therapy.

关 键 词：TOLL样受体 土拨鼠肝炎病毒 原代肝细胞 转基因小鼠 

分 类 号：R512.6[医药卫生—内科学]