三种IFN-α抗病毒活性新方法的比较  

Comparison among three new ways of antiviral activity about interferon alpha

作　　者：卢年芳[1] 吴莹[1] 唐霓[1] 郑瑞强[2] 朱亚彬[2] 闫歌[1] 张秉强[1] 黄爱龙[1] 

机构地区：[1]重庆医科大学病毒性肝炎研究所国家级重点实验室,重庆市400010 [2]扬州大学医学院附属江苏省苏北人民医院,江苏省扬州市225001

出　　处：《世界华人消化杂志》2005年第16期1964-1969,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30300298~~

摘　　要：目的:体外研究不同亚型干扰素-α(IFN-α2b,IFN-α2a,IFN-α1b)抗HBV活性和信号分子基因应答水平的差异性,以探讨用信号分子作为一种新的抗病毒评价标准的可能性.方法:0.5,1,2,4,8MU/LIFN-α2b,IFN-α2a,IFN-α1b作用于2.2.15细胞后,用Abbott诊断试剂盒分别检测上清中HBsAg,HBeAg的含量,并计算其抑制率;1MU/LIFN-α2b,IFN-α2a,IFN-α1b作用于HepG2细胞后,利用RT-PCR及Westernblotting方法检测胞内STAT1,IFNARmRNA及蛋白表达水平的差异性.结果:IFN-α对HBsAg,HBeAg抑制率随药物浓度增加而增强,当IFN-α浓度为0.5,1MU/L时,三组不同亚型IFN-α对HBsAg,HBeAg抑制率无统计学差异.当浓度为2,4,8MU/L时,IFN-α1b组对HBsAg,HBeAg抑制率明显较IFN-α2b,IFN-α2a组高(HBsAg:F=4.51,6.23;HBeAg:F=3.11,4.72,P<0.05),而IFN-α2b,IFN-α2a组间无统计学差异.RT-PCR和Westernblotting结果1Mu/L时,IFN-α1b处理组IFNAR,STAT1mRNA及蛋白表达水平均较IFN-α2b组高,两组比较无统计学差异.IFN-α1b,IFN-α2b处理组IFNAR,STAT1mRNA及蛋白表达水平明显较IFN-α2a组高,均存在统计学差异(mRNA:F=5.26,15.6;蛋白:F=17.7,20.1,P<0.05).结论:IFN-α1b,IFN-α2b抗HBV活性较强,IFN-α2a较弱.用信号分子STAT1,IFNAR来评价IFN-α抗病毒活性则更为敏感.AIM： To investigate the antiviral activity on Hepatitis B Virus and the gene responses of interferon alpha with different subtypes in vitro, and to assess the feasibility of using the signal transduction molecules as a new standard for evaluating the antiviral activities of IFN-α（subtypes. METHODS： After 2.2.15 cells were respectively treated with IFN-α（2b, IFN-α（ 2α, IFN-α 1 b of varied concentrations （0.5,1,2, 4, 8 MU/L）, the contents of HBsAg and HBeAg in the supernatant were measured by Abbot kit and the inhibitory rates on HBsAg and HBeAg were calculated. After HepG2 cells were treated with 1 MU/L IFN-α（2b, I FN-α2b, IFN-α 1b, the levels of STAT1, IFNAR mRNA and protein of were detected by reverse transcription polymerase chain reaction （RT-PCR） and Western blotting. RESULTS： The inhibitory rate of IFNα2b, IFN-α2a, IFNα 1 b on HBsAg and HBeAg showed no statistical differences at the concentration of 0.5 or 1 MU/L. At the concentrations of 2, 4 and 8 MU/L, the inhibitory rate of IFN-αlbwas significantly higher than that of IFN-α 2b or IFN-α 2a （HBsAg： F= 4.51,6.23; HBeAg： F= 3.11,4.72, all P〈0.05）, while there was no significant difference between IFN-α 2b and IFN-α 2a. The levels of IFNAR, STAT1 mRNA and protein expressionwere slightly higher in IFN-α I b group than that in IFN-α 2b group. However, the levels of mRNA and protein expression in IFN-αI b or IFN-α2b group mark- edly higher than that in IFN-α2a group （mRNA： F= 5.26, 15.6; protein： F= 17.7, 20.1, all P〈0.05）. CONCLUSION： IFN-α I b and IFN-α 2b have stronger antiviral activity on HBV than IFN-α 2a. The signal transduction molecules （STAT1, IFNAR） are more sensitive in evaluating the antiviral activity of IFN-α.

关 键 词：干扰素-Α 乙型肝炎病毒 信号分子 IFN-Α2B 抗病毒活性 IFN-Α2A BLOTTING 差异RT-PCR 蛋白表达水平 Western 

分 类 号：R96[医药卫生—药理学]