HBx蛋白抑制乙型肝炎病毒诱导肝细胞表达Ⅰ型干扰素的研究  

HBx protein inhibits expression of type Ⅰ interferon in hepatocytes induced byhepatitis B virus

作　　者：潘颖 杨凯 陈谨[1] 孙蓓蓓[2] 田平平 张发苏 PAN Ying;YANG Kai;CHEN Jin;SUN Beibei;TIAN Pingping;ZHANG Fasu(Department of Medical Tech-nology,Anhui Medical College,Hefei 230601,China;Department of Laboratory Medicine,the Second Affiliated Hospital of Anhui Medical University,Hefei 230601,China)

机构地区：[1]安徽医学高等专科学校医学技术学院,合肥230601 [2]安徽医科大学第二附属医院检验科,合肥230601

出　　处：《中国免疫学杂志》2024年第2期252-255,共4页Chinese Journal of Immunology

基　　金：安徽省自然科学基金项目(2108085MH301);安徽省高校省级自然科学研究重点项目(KJ2020A0861)。

摘　　要：目的:研究乙型肝炎病毒(HBV)对Ⅰ型干扰素表达的影响,并进一步探讨相关机制。方法:以HepG2细胞和稳转HBV基因组的HepG2.2.15细胞为模型,转染针对乙型肝炎病毒X(HBx)基因的小干扰RNA(siRNA)[HBx-siRNA]至HepG2.2.15细胞,转染HBx蛋白表达质粒pEGFP-HBx至HepG2细胞,荧光定量PCR技术分析各细胞中的IFN-α、IFN-β mRNA含量,荧光显微镜观察HepG2细胞内质粒pEGFP-HBx的表达,Western blot技术分析各细胞中HBx和p-IRF3蛋白含量变化,以探讨HBV通过HBx蛋白对Ⅰ型干扰素表达的影响。结果:IFN-α和IFN-β mRNA在HepG2.2.15细胞中的含量略高于HepG2细胞,差异无统计学意义(P>0.05);RNA干扰抑制HepG2.2.15细胞中HBx蛋白的表达后,细胞中IFN-α、IFN-β mRNA含量显著升高,转染pEGFP-HBx的HepG2细胞中Ⅰ型干扰素的基因表达量和p-IRF3的蛋白表达量均降低。结论:HBV通过HBx蛋白抑制IRF3蛋白的活化,从而抑制Ⅰ型干扰素的表达。Objective:To investigate the effect of hepatitis B virus(HBV)on the expression of typeⅠinterferon and its possi-ble mechanism.Methods:HepG2 cells and HBV stably transfected HepG2.2.15 cells were used as the model,cells were transfected with pEGFP-HBx plasmid or HBx-siRNA,the levels of IFN-αand IFN-βmRNA in cells were detected by fluorescence quantitative PCR,and the expression of pEGFP-HBx plasmid in HepG2 cells was observed by fluorescence microscope,the changes of HBx and p-IRF3 protein in cells were analyzed by Western blot,to investigate the effect of HBV on the expression of typeⅠinterferon.Results:The levels of IFN-αand IFN-βmRNA in HepG2.2.15 cells were slightly higher than those in HepG2 cells,there was no significant dif-ference(P>0.05);after silencing HBx expression by using RNA interference in HepG2.2.15 cells,the levels of IFN-αand IFN-βmRNA were significantly increased.Moreover,the levels of typeⅠinterferon genes expression and p-IRF3 protein expression in HepG2 cells transfected with pEGFP-HBx were both decreased.Conclusion:HBV inhibits the expression of typeⅠinterferon by reg-ulating the activation of IRF3 protein through HBx protein.

关 键 词：乙型肝炎病毒 HBX蛋白 Ⅰ型干扰素 IRF3蛋白 

分 类 号：R392.1[医药卫生—免疫学]