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目的探讨细胞内病毒识别受体RIG-I激活后对EV71复制的影响。方法分别用Poly(I:C)和表达RIG-I N端的质粒转染RD细胞,24 h后感染EV71,感染24 h后收获细胞和上清。提取细胞RNA并反转录为cDNA,Real-time PCR测定细胞中RIG-I与IFN-β的mRNA的表达水平和EV71的RNA水平。用免疫印迹的方法检测RIG-I和EV71的蛋白表达水平。将感染病毒的细胞上清进行病毒滴度的测定。结果 RIG-I被激活之后RIG-I与IFN-β的mRNA的表达水平上升,并且RIG-I的蛋白表达可以被检测到。感染细胞中的EV71的RNA水平和病毒蛋白表达水平下降,并且病毒滴度显著降低。结论 RIG-I的激活以及其诱导的IFN-β对EV71的复制有抑制效应。
Objective To investigate the effect of intracellular viral recognition receptor RIG-I on EV71 replication. Methods RD cells were transfected with Poly (I: C) and RIG-I N-terminal plasmids, respectively. EV71 cells were infected 24 hours later and cells and supernatant were harvested 24 hours after infection. The cellular RNA was extracted and reverse transcribed into cDNA. The mRNA expression of RIG-I and IFN-β in the cells and the RNA level of EV71 were determined by Real-time PCR. Western blotting was used to detect the protein expression of RIG-I and EV71. The virus-infected cell supernatant was assayed for virus titer. As a result, the mRNA level of RIG-1 and IFN-β was increased after RIG-I was activated, and the protein expression of RIG-1 was detected. The level of RNA and the expression of viral proteins in EV71 in infected cells decreased, and the virus titer was significantly reduced. Conclusion The activation of RIG-I and its induction of IFN-β have an inhibitory effect on the replication of EV71.