6-磷酸葡萄糖脱氢酶在乙型肝炎病毒复制中的作用及其机制

Role of glucose-6-phosphate dehydrogenase in hepatitis B virus repfication and its possible mechanism of action

目的探讨6-磷酸葡萄糖脱氢酶（G6PD）在乙型肝炎病毒（HBV）复制中的作用及其可能机制。方法采用组织芯片、实时定量PCR和Westernblot技术对HBV阳性和阴性的肝组织和细胞株进行G6PD表达的差异性进行分析；采用siRNA技术对HepG2．2．15细胞的G6PD基因进行敲减培养48h。采用化学发光法对培养上清液中的HBsAg和HBeAg进行定量检测；实时定量PCR进行HBVDNA和I型干扰素（IFN）及其下游的干扰素刺激基因检测。两组间的比较采用f检验。结果G6PD在HBV阳性的肝组织和细胞中的表达显著高于阴性组，染色强度及免疫组织化学得分为89．69±54．92对比31．90±18．62，P〈0．05。siRNA干扰HepG2．2．15细胞表达G6PD后，培养上清液中的HBVDNA、HBsAg、HBeAg定量水平均明显下降，差异具有统计学意义；IFNCα1、IFNβ1以及下游的5个干扰素刺激基因（OAS1、ISG15、OAS3、EIF2α、PKR）的mRNA水平均明显升高，差异具有统计学意义，尸值均〈0．05。结论G6PD对HBV复制具有重要的促进作用，其调控HBV复制的机制可能与I型干扰索途径相关。

Objective To investigate the role of glucose-6-phosphate dehydrogenase （G6PD） in hepatitis B virus （HBV） replication and its possible mechanism of action. Methods Tissue microarray, quantitative real- time PCR, and Western blot were performed to analyze the differences in G6PD expression levels in the HBV- positive and HBV-negative liver tissues, HepG2.2.15 cells, and HepG2 cells. The siRNA transfection technique was used to knock down G6PD gene in HepG2.2.15 cells for 48 hours. Chemiluminescence was used for HBsAg and HBeAg quantification in supernatant, and quantitative real-time PCR was used to measure HBV DNA, type I interferon （IFN）, and downstream IFN-stimulated genes. The t-test was used for comparison between groups. Results G6PD expression was significantly upregulated in the HBV-positive liver tissues and cells compared with HBV-negative liver tissues and cells, and the stain intensity and immunohistochemical scores were 89.69±54.92 and 31.90±18.62, respectively （P 〈 0.05）. After G6PD expression in HepG2.2.15 cells was interfered by siRNA, the quantitative levels of HBV DNA, HBsAg, and HBeAg in supernatant were reduced significantly, and the mRNA expression levels of IFNαl, IFN[31, and five downstream IFN-stimulated genes （OAS1, ISG15, OAS3, EIF2α, and PKR） increased significantly （allP 〈 0.05）. Conclusion G6PD plays a vital role in HBV replication, and its mechanism of action in regulating HBV replication may be related to type I IFN signaling pathway.