ISGF3:IFN-α抗乙型肝炎病毒信号转导机制的重要因子  

ISGF3, a critical factor of the IFN-α pathway in the antiviral action of HBV

作　　者：张权[1] 魏来[1] 王燕[1] 

机构地区：[1]北京大学人民医院北京大学肝病研究所,100044

出　　处：《中华实验和临床病毒学杂志》2005年第2期110-113,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家 973计划项目 (G1990 5 410 6)

摘　　要：目的　研究干扰素 α(IFN α)抗乙型肝炎病毒(HBV)的信号转导机制。方法　用定量聚合酶链反应(PCR)检测IFNα2b处理前、后的HepG2 2 15细胞上清的乙型肝炎病毒脱氧核糖核酸(HBVDNA)水平。半定量逆转录聚合酶链反应(RT PCR)检测IFNα2b处理后不同时点的HepG2 2 15细胞和其母源细胞HepG2 细胞内信号转导和转录活化因子1(STAT1)、STAT2、干扰素刺激基因因子3γ(ISGF3γ)、2′5′寡腺苷酸合成酶(2′5′OAS)、RAN依赖蛋白激酶(PKR)的mRNA表达水平。WesternBlot检测ISGF3γ的蛋白质表达水平。并用染料木黄酮阻断IFNα2b信号转导后再次检测上述指标。结果　IFNα2b处理8h后,HepG2 2 15细胞上清HBVDNA平均减少0 72log 10copies ml,而加染料木黄酮后则无减少。IFNα2b处理后,HepG2 和HepG2 2 15细胞中STAT1、STAT2、ISGF3γ、2′5′OAS和PKRmRNA水平明显升高;加染料木黄酮后前3个因子mRNA水平仍然能检测到,而2′5′OAS和PKRmRNA则受到抑制。WesternBlot检测也提示IFNα2b处理后ISGF3γ蛋白质水平升高,加染料木黄酮后其表达受到抑制。结论　Janus酪氨酸激酶(JAK) STAT途径在IFN α抗HBV中发挥主要作用,而ISGF3是该途径的一个重要因子。Objective To study the mechanism of signal transduction in anti-HBV action of IFN-α. Methods The HBV DNA in HepG 2.2.15 cell line supernatant with/without IFNα-2b were monitored by fluorescence real-time quantitive PCR. STAT1,STAT2,ISGF3γ,PKR, 2′5′-OAS mRNA levels from HepG 2 and HepG 2.2.15 cell lines that were treated with/without IFNα-2b at different times were detected by semi-quantitive RT-PCR. And the ISGF3γ protein was detected by Western blot. Then, these items were detected again after inhibiting the JAK-STAT pathway with genistein.Results The HBV DNA in 2215 supernatant that were treated with IFNα-2b for 8 hours decreased 0.72 log 10 copies/ml. But the basal levels of DNA in cells pretreatred with genistein, followed by IFN-α2b did not decrease. The STAT1,STAT2,ISGF3γ,2′5′-OAS,PKR mRNA levels were upregulated by IFN-α2b. The same phenomena were observed with STAT1,STAT2,ISGF3γ mRNA when pretreated with genistein then treated with IFN-α2b,but the levels of 2′5′-OAS, PKR mRNA were decreased in this situation. The expression of the protein of ISGF3γ was also augmented by IFNα-2b,and was blocked by genistein. Conclusion The JAK-STAT pathway seems to be a critical pathway in IFNα-2b action against HBV, and ISGF3 is most probably a key factor of the route.

关 键 词：信号转导机制 重要因子 抗乙型肝炎病毒 HepG2.2.15细胞 乙型肝炎病毒脱氧核糖核酸 半定量逆转录聚合酶链反应 IFNΑ-2B 乙型肝炎病毒(HBV) 聚合酶链反应(PCR) Western 染料木黄酮 BLOT检测 mRNA水平 细胞内信号转导 

分 类 号：R96[医药卫生—药理学]