MyD88抑制乙型肝炎病毒复制依赖活化NF-κB信号通路  

Inhibition of hepatitis B virus replication by MyD88 is mediated by nuclear factor-kappa B activation

作　　者：林珊珊[1] 邬敏[1,2] 徐杨[1] 熊炜[1] 张小楠[1,2] 易志刚[1] 袁正宏[1,2,3] 

机构地区：[1]复旦大学上海医学院医学分子病毒学重点实验室,上海200032 [2]上海市公共卫生中心科研中心 [3]复旦大学生物医学研究院

出　　处：《微生物与感染》2007年第2期73-77,106,共6页Journal of Microbes and Infections

摘　　要：目的研究髓样细胞分化蛋白(MyD88)抗乙型肝炎病毒(HBV)效应的作用机制。方法构建MyD88的截短突变体,获得核因子kappa B(NF-κB)超抑制剂IkBa-SR或者NF-κB信号通路激活剂IKKα/IKKβ的表达质粒,分别与HBV复制型质粒瞬时转染Huh7细胞,检测细胞上清液中HBeAg,HBsAg的表达以及胞质中HBV复制中间体DNA的含量,并以NF-κB依赖的荧光素酶报道系统检测它们活化NF-κB的程度。结果MyD88全长蛋白和2个截短突变体M(1-151)、M(151-296)活化NF-κB的程度与其抑制HBV蛋白以及复制中间体DNA合成的能力相一致。与空载相比,表达NF-κB信号通路激活剂IKKα/IKKβ的质粒共同瞬转细胞后,转染MyD88和HBV表达质粒的细胞中NF-κB的通路明显活化,同时HBV core蛋白的合成显著降低;而NF-κB的超抑制剂IκBα-SR共同瞬转的细胞中core蛋白的表达量显著增加,检测细胞培养上清液中HBeAg和HBsAg及胞质中HBV复制中间体DNA的合成,得到相似结果。结论NF-κB信号通路的活化在MyD88抑制HBV复制中发挥了关键作用。Objective To investigate the mechanism of MyD88 inhibition of hepatitis B virus （HBV） replication. Methods Two truncated versions of MyD88, M （1-151） and M （152-296） were constructed. Plasmid IκBα-SR encoding NF-κB super-repressor or plasmid IKKa/IKI~ which can strongly activate NF-κB signaling were used. Huh7 cells were transiently transfected with HBV dimer, MyD88 or the above plasmids. Assays of HBsAg, HBeAg, core protein, HBV DNA and NF-κB activation were performed. Results The expression of the full length MyD88, and the two truncated forms, M （ 1-151 ） and M （ 152-296）, showed different induction patterns of NF-κB activity; patterns that are consistent with their inhibitory effect on viral protein synthesis and core particle-associated HBV DNA replication. Similarly, the expression of MyD88 resulted in a significant reduction of core protein and the co-expression of IKKα/IKKβ dramatically inhibited the core protein level. Furthermore, the co-expression of IκBα-SR dramatically restored the core protein level. Similar results were also observed for HBeAg, HBsAg and HBV core particle DNA. Conclusion Results from these studies support a role of NF-κB signaling activation in HBV viral replication and suggest a novel mechanism for the inhibition of HBV replication by MyD88.

关 键 词：乙型肝炎病毒 髓样细胞分化蛋白 抗病毒活性 核因子KAPPA B 

分 类 号：R373[医药卫生—病原生物学]