RNA干扰上调天然免疫  

RNAi Enhances Innate Immunity

作　　者：孟忠吉[1] 张永红[2] 李东[1] 汤守兵[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,湖北十堰442000 [2]湖北医药学院附属太和医院中西医结合科,湖北十堰442000

出　　处：《湖北医药学院学报》2013年第4期281-285,共5页Journal of Hubei University of Medicine

基　　金：湖北省自然科学基金(2010CDZ036);湖北医药学院优秀中青年科技创新团队项目(2011CXX02)

摘　　要：目的:研究特异性RNA干扰对于天然免疫的上调作用。方法:分别分离WHV感染的土拨鼠和WHV转基因小鼠原代肝细胞,转染针对WHV、小鼠β-actin和GAPDH的siRNA。Northern blot和Real-time RT-PCR分别检测原代肝细胞中WHV、MxA、MHCⅠ、β-actin、GAPDH、IFN-β和IP-10的mRNA水平。结果:针对WHV、小鼠β-actin和GAPDH的siRNA抑制靶基因mRNA伴随MxA、MHCⅠ、IP-10等干扰素刺激基因的上调表达。这种干扰素刺激基因的上调表达只有在靶基因被特异siRNA降解时出现,呈剂量和沉寂效应依赖性,而且可以被siRNA特异性抑制剂所阻断。结论:RNAi上调天然免疫可能增强siRNA的抗病毒效应,有可能在抗病毒RNAi研究中发挥重要作用。Objective To investigate the effect of RNAi on innate immunity.Methods Primary hepatocytes were isolated from WHV infected woodchucks and WHV transgenic mice,and transfected with siRNAs targeting WHV,mouse β-actin and GAPDH.The mRNA level of WHV,MxA,MHCⅠ,β-actin,GAPDH,IFN-β and IP-10 in primary hepatocytes were determined by Northern blotting and real-time RT-PCR,respectively.Results All the siRNAs led to depletion of their targeting transcripts accompanied with increased level of IFN-β,IP-10,MxA,or MHCⅠ.The ISGs upregulation was only occurred in the effective RNAi with targeting mRNA degradation,dose-and RNAi effect dependently.The RNAi-directed ISGs upregulation could be blocked with siRNA specific inhibitor.Conclusion RNAi directed enhancement of innate immunity might amplify antiviral effects of siRNAs and play an important role in the anti-viral study.

关 键 词：RNA干扰 土拨鼠肝炎病毒 原代肝细胞 天然免疫 干扰素刺激基因 

分 类 号：R392.1[医药卫生—免疫学]