血红素加氧酶-1通过诱导抗病毒蛋白的表达增强IFN-α抗HBV效应  

Heme oxygenase-1 enhances the anti-HBV effect of IFN-α by inducing the expression of antiviral proteins

作　　者：笪蔚 王琴[1] 魏安邦 张浩[1] 汪任冰 刘倩 周强[1] Da Wei;Wang Qin;Wei Anbang;Zhang Hao;Wang Renbing;Liu Qian;Zhou Qiang(Dept of Laboratory Medicine,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601;Dept of Pediatrics,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601)

机构地区：[1]安徽医科大学第二附属医院检验科,合肥230601 [2]安徽医科大学第二附属医院儿科,合肥230601

出　　处：《安徽医科大学学报》2024年第2期324-330,共7页Acta Universitatis Medicinalis Anhui

基　　金：安徽省转化医学研究院科研基金项目(编号:2021zhyx-C47);安徽省高校自然科学研究重点项目(编号:2023AH053169)。

摘　　要：目的探讨血红素加氧酶-1(HO-1)对HBV复制的作用及HO-1联合α-干扰素(IFN-α)的抗病毒效应。方法以HepG2.2.15细胞和HBV 1.3质粒转染HepG2细胞即HepG2-HBV1.3为HBV复制细胞模型;血红素(Hemin)分别处理HepG2.2.15和HepG2-HBV1.3细胞,诱导HO-1表达;CCK-8评估Hemin对HepG2、HepG2.2.15的毒性作用;化学发光法分析Hemin处理组及si-HO-1等实验组上清液中HBsAg、HBeAg;RT-qPCR分析HO-1、IFN-β、HBV-DNA;Western blot分析IRF-3、JAK/STAT信号通路中相关分子的表达;Hemin联合IFN-α处理HepG2.2.15,监测HO-1是否具有协同IFN-α抗病毒效应。结果Hemin剂量依赖性诱导HO-1,HO-1被诱导后发挥显著的抗HBV效应,同时IFN-β、IRF-3及JAK/STAT信号通路中IRF-9、MxA的表达均增加。沉默HO-1表达能逆转Hemin诱导组的抗病毒效应,同时I型干扰素IFN-β也呈现低表达,JAK/STAT信号通路中的IRF-9、MxA的表达也被抑制。Hemin联合IFN-α发挥更强的抗病毒作用。结论HO-1能够发挥抗HBV效应,这种效应可能是增加IRF-3的磷酸化诱导I型干扰素表达来激活JAK/STAT信号通路发挥抗病毒效应;HO-1可以协同IFN-α发挥抗病毒作用。Objective To investigate the role of heme oxygenase-1(HO-1) on HBV replication and the antiviral effect of HO-1 combined with α-interferon(IFN-α).Methods HepG2.2.15 cells and HBV1.3-transfected HepG2 cells(HepG2-HBV1.3) were used as HBV replicating cell models;Hemin treated HepG2.2.15 and HepG2-HBV1.3 cells,to induce the expression of HO-1 molecules.CCK-8 method was used to assess the toxic effects of Hemin on HepG2 and HepG2.2.15;chemiluminescence method was used to analyze HBsAg and HBeAg in the supernatants of Hemin-treated group and si-HO-1 and other experimental groups;RT-qPCR was used to analyze HO-1,IFN-β and HBV-DNA;Western blot was used to analyze the expression of IRF-3 and the expression of related molecules in the JAK/STAT signaling pathway;Hemin combined with IFN-α treated HepG2.2.15 to monitor whether HO-1 had synergistic IFN-α antiviral effect.Results Hemin dose-dependently induced HO-1,and HO-1 was induced to exert a significant anti-HBV effect,while the expression of IFN-β,IRF-3,and IRF-9 and MxA,downstream molecules of the JAK/STAT signaling pathway,were all increased.Silencing HO-1 expression reversed the antiviral effect in the Hemin-induced group,and at the same time,type I interferon IFN-β showed low expression,and the expression of IRF-9 and MxA in the JAK/STAT signaling pathway was inhibited as well.Hemin combined with IFN-α exerted stronger antiviral effects.Conclusion HO-1 can exert an anti-HBV effect,which may be due to increased phosphorylation of IRF-3 to induce type I interferon expression and thus activate the JAK/STAT signaling pathway to exert an antiviral effect;HO-1 can synergize with IFN-α to exert an antiviral effect.

关 键 词：血红素 血红素加氧酶-1 JAK/STAT IFN-Β 

分 类 号：R512.62[医药卫生—内科学]