干扰素α抗病毒活性的实验研究  

An experimental study on antiviral effects of IFN α

作　　者：卢年芳[1] 黄爱龙[2] 唐霓[2] 郑瑞强[1] 林华[1] 朱亚彬[1] 吴莹[2] 陶鹏[2] 

机构地区：[1]扬州大学医学院附属医院,江苏省苏北人民医院,225001 [2]重庆医科大学病毒性肝炎研究所

出　　处：《中华肝脏病杂志》2005年第12期892-896,共5页Chinese Journal of Hepatology

基　　金：国家自然科学基金(30300298)

摘　　要：目的 比较不同亚型干扰素α(IFN α2b、IFN α2a、IFN α1b)对 JAK-信号转导活化转录 因子(STAT)通道中重要信号传导分子 STAT1、STAT2、干扰素α受体(IFNAR)、蛋白激酶(PKR)、核 糖核酸酶 L(R Nase L)表达的影响，研究其抗病毒活性的差别，并评价 IFN α抗病毒活性的关键性信号传 导分子。方法 1000 U/ml 的不同亚型 IFN α分别作用于 HepG2细胞后，用逆转录聚合酶链反应(RT- PCR)方法检测 HepG2细胞内 STAT1、STAT2、IFNAR、PKR、RNase L mRNA 表达水平。用 western blot 方法检测细胞内 STAT1及 IFNAR 蛋白表达水平。结果 RT-PCR 的实验结果：(1)IFN α1b 处理组 IFNAR、 STAT1、STAT2 mRNA 的表达水平较 IFN α2b 组高，两组比较差异无统计学意义。IFN α1b、IFN α2b 处 理组 IFNAR、STAT1、STAT2 mRNA 水平明显较 IFN α2a 组高，差异均有统计学意义，F 值分别为5．26、 15．6、4．67，P 值均＜0．05。(2)IFN α1b、IFN α2b、IFN α2a 处理后的 PKR 表达水平相似，3组比较差异 无统计学意义。(3)RNase L 表达量极少，无法比较不同处理组间 RNase L mRNA 表达水平的差异性。Western blot 实验结果：(1)IFN α1b 处理组 IFNAR、STAT1蛋白水平较 IFN α2b 处理组高，两者比较差异无统计 学意义，IFN α1b、IFN α2b 处理组 IFNAR、STAT1蛋白水平较 IFN α2a 处理组明显升高，差异有统计学 意义，F 值分别为17．7、20．1，P 值均＜0．05。结论 IFN α2b、IFN α2a、IFN α1b 均可促进 JAK-STAT 信号通道中 STAT1、STAT2、IFNAR mRNA 及蛋白表达，IFN α1b 和 IFN α2b 作用较强，IFN α2a 作用 较弱。初步表明，IFN α1b、IFN α2b 的抗病毒活性较 IFN α2a 强，IFNAR、STATl、STAT2可作为评价 IFN α抗病毒活性的关键性指标，而 PKR、RNase L 能否作为评价指标有待进一步的实验证实。Objectives To investigate the effects of different subtypes IFN α（IFN α2b, IFNα2a, and IFN α1b） transduction molecular STAT1, STAT2, IFNAR, PKR, and RNase L, and to study the differences of their antiviral effects and to evaluate the key signaling transduction molecules. Methods （1） After HepG2 cells were treated with IFN α2b, IFN α2a, or IFN α1b, the mRNA levels of STAT1, STAT2, IFNAR, PKR, and RNase L were detected by RT-PCR. （2） After HepG2 cells were treated with 1000 U/ml IFN α2b, IFN α2a, or IFN α1b, the protein expression levels of STAT1 and IFNAR were examined by Western blot. Results RT-PCR results： （1） IFNAR, STAT1, and STAT2 mRNA expression levels were slightly higher in the IFN α1b group than those in the IFN α2b group （P 〉 0.05）. The mRNA expression levels in IFN α1b or IFN α2b groups were significantly higher than in the IFN α2a group （P 〈 0.05）. （2） The PKR mRNA expression showed no significant differences among IFN α1b, IFN α2b, and IFN α2a groups. （3） The RNase L mRNA expression was very weak. We could not compare the differences of the RNase L mRNA levels in different groups by RT-PCR. Western blot results： （1） The IFNAR, and STAT1 protein expressions were greatly upregulated after IFN a induction compared with the untreated group （P 〈 0.05）. （2） The IFNAR, and STAT1 protein expression levels in IFN a lb group were slightly higher than the IFNα2b group. IFNAR, and STAT1 protein levels of IFNa lb or IFNa2b group were significantly higher than IFNα2a group （P 〈 0.05）. Conclusion STAT1, STAT2, IFNAR mRNA and protein expressions could all be markedly upregulated after IFNa treatment. Effects of IFNα lb or IFNα 2b were greatly stronger than IFNα2a. The PKR mRNA expression also was greatly upregulated after IFNα treatment. Expression levels of PKR in IFNα1b, IFNα2b, and IFNα 2a groups were all similar. The mRNA level results were consistent with the protein level results. Our results showed that the antiviral activi

关 键 词：干扰素Α 信号传导 治疗学 

分 类 号：R96[医药卫生—药理学]