MxA蛋白对HepG2.2.15细胞HBV复制的影响

The inhibition of HBV replication by MxA protein in HepG2.2.15 cells

目的分析比较α干扰素(IFN-α)、抗黏液病毒A(MxA)蛋白及IFN-α联合MxA蛋白对HepG2.2.15细胞中乙型肝炎病毒(HBV)复制的影响。方法以肝胚瘤细胞株HepG2.2.15细胞为细胞模型,以IFN-α、基因转染表达MxA蛋白及IFN-α与MxA蛋白联合等方式处理细胞,Western blot法检测MxA蛋白在细胞中表达;ELISA法和实时荧光定量PCR技术分析细胞上清HBV抗原和细胞外HBV DNA的表达。结果 IFN-α处理后,HepG 2.2.15细胞分泌的HBV抗原未见显著减少,而MxA蛋白单处理组HepG2.2.15细胞分泌的HBV抗原显著减少(P<0.05);联合处理组具有抑制HBV抗原分泌的作用(P<0.01),但3种处理组均不能抑制细胞外HBV DNA的表达。结论在体外细胞模型中,IFN-α单独处理不能有效抑制HepG2.2.15细胞中HBV复制及抗原分泌;MxA蛋白能够抑制HBV抗原分泌;IFN-α与MxA蛋白联合作用具有抑制HBV抗原分泌的作用,但不影响细胞外HBV DNA复制。

Objective To analyse IFN-α,MxA protein and IFN-α combined with MxA protein on HBV replication in HepG2.2.15 cells.Methods HepG2.2.15 cells were treated with IFN-α,transfected with pcDNA3.1-Flag-MxA（wild-type） plasmid,and IFN-α combined with MxA protein.The expression of MxA protein in HepG2.2.15 cells was analysed by Western blot.Meanwhile,HBV DNA was detected by real-time PCR and HBsAg/HBeAg were analysed by ELISA assay.Results MxA protein was expressed in HepG2.2.15 cells transfected with pcDNA3.1-Flag-MxA（wild-type）.When the cells were treated with MxA protein,compared with the control group cells,the expression of HBsAg and HBeAg in the cell culture supernantants decreased（P0.05）.Respectively,the amounts of HBsAg and HBeAg in combination group cells decreased significantly（P0.01）.However,the extracellular HBV DNA in the three groups was not inhibited significantly.Conclusion In vitro,IFN-α treatment alone can not inhibit HBV replication in HepG2.2.15 cells.MxA protein can inhibit the secretion of HBV antigen,and IFN-α combined with MxA protein has more significant role in inhibiting HBV replication,but not affect the extracellular HBV DNA.