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目的探讨肠道病毒71型(enterovirus 71,EV71)感染对Ⅰ型干扰素信号通路中核转位复合物干扰素刺激基因因子3(ISGF3)核转位的影响。方法 Western印迹检测VR1432感染VeroE6对干扰素诱导的STAT1和STAT2磷酸化的影响。将pEGFP-C1-STAT1重组质粒转染Vero E6细胞,随后进行EV71感染以及IFN-α2b刺激,将细胞进行亚细胞分离检测ISGF3的核转位情况。结果 VR1432以感染复数(MOI)为1感染Vero E6细胞后未影响干扰素刺激的STAT1和STAT2的磷酸化;亚细胞分离后Western印迹检测磷酸化的STAT1,结果显示VR1432感染后,p-STAT1在细胞核内表达水平明显降低,说明EV71感染抑制了p-STAT1的核转位。结论 EV71感染抑制Ⅰ型干扰素信号通路中ISGF3(STAT1/STAT2/IRF9)的核转位,从而影响干扰素效应途径中抗病毒蛋白的表达。
Objective To investigate the effect of enterovirus 71 (EV71) infection on the nuclear translocation of interferon stimulating factor 3 (ISGF3) in the type Ⅰ interferon signaling pathway. Methods Western blotting was used to detect the effect of VR1432 infection on Vero E6 on IFN-induced STAT1 and STAT2 phosphorylation. The recombinant plasmid pEGFP-C1-STAT1 was transfected into Vero E6 cells, followed by EV71 infection and IFN-α2b stimulation. The cells were sub-cultured for the detection of nuclear translocation of ISGF3. Results The expression of STAT1 and STAT2 in Vero E6 cells infected with VR1432 at a multiplicity of infection (MOI) of 1 did not affect the phosphorylation of STAT1 and STAT2 stimulated by interferon. After subcellular isolation, phosphorylated STAT1 was detected by Western blotting. In the nucleus, the expression of p-STAT1 was significantly decreased, indicating that EV71 infection inhibited the nuclear translocation of p-STAT1. Conclusion EV71 infection inhibits the nuclear translocation of ISGF3 (STAT1 / STAT2 / IRF9) in the type I interferon signaling pathway and thus affects the expression of antiviral proteins in the IFN-a pathway.