戊型肝炎病毒开放读码框3蛋白抑制肿瘤坏死因子-α介导肝细胞p65的核转移

Open reading frame 3 protein of hepatitis E virus suppresses TNF-α-mediated NF-κB activation in hepatocyte

目的:研究Ⅰ型和Ⅳ型戊型肝炎病毒(HEV)开放读码框3蛋白(ORF3)对肝细胞系L02核因子-kap-pa B(Nuclear Factor kappa B,NF-κB)通路的影响。方法:分别用未转染质粒、转染Ⅰ型和Ⅳ型ORF3真核表达质粒及空载质粒的L02细胞,48小时后加入TNF(肿瘤坏死因子)-α刺激,采用免疫荧光观察细胞内p65的核转运,并用western blot检测p65蛋白的表达。结果:未转染质粒的L02细胞加入TNF-α刺激后,p65由胞浆转入胞核;转染空载的L02细胞加入TNF-α刺激后,与未转染的L02相似,p65由胞浆转入胞核;表达Ⅰ型和Ⅳ型ORF3蛋白的细胞,加入TNF-α刺激后,p65未见明显核转移。结论:Ⅰ型和Ⅳ型ORF3蛋白可抑制TNF-α介导的p65核转移。

Objective：To study the influence of ORF3 protein of type Ⅰand type Ⅳ hepatitis E virus（HEV） on NF-κB pathway in hepatocyte.Methods：Each plasmids（with or without full-length of orf3 gene） were transfected into L02 cells.After 48 hours,the cells were stimulated with TNF-α.Immonofluorescence assay and western blot analysis were used to observe the translocation of p65.Results：In our study,we could observe that p65 translocated from cytoplasm to nuclear,neither L02 transfected without plasmid nor with control plasmid after stimulate of TNF-α.However,there were no obvious nuclear translocation in ORF3 transfected cells after stimulate of TNF-α.Conclusion： ORF3 protein of hepatitis E virus suppresses TNF-α-mediated NF-κB p65 activation in hepatocyte.