摘要
目的:本实验以小鼠系膜细胞为研究对象,IFN-γ为刺激物,通过检测JAK/STAT信号途经的激活及HMGB1mRNA及蛋白表达,探讨γ干扰素刺激系膜细胞HMGB1表达上调的可能机制。方法:常规培养小鼠系膜细胞(Mousemesangial cell,MMC),无血清培养基同步化后分为正常对照组、IFN-γ(500 U/ml)刺激组和INF-γ+AG490(INF-γ500 U/ml+AG490 200μmol/ml)组。Western blot检测JAK、STAT1和HMGB1蛋白的表达变化,RT-PCR检测HMGB1mRNA的表达变化。结果:Western blot结果显示IFN-γ能够促进小鼠系膜细胞JAK、STAT1磷酸化和STAT1核转位,并呈时间依赖性;AG490能够降低IFN-γ介导的JAK1、JAK2、STAT1的活化,并呈时间依赖性。IFN-γ能够上调系膜细胞中HMGB1mRNA及蛋白表达,并随着刺激时间延长,其相对表达量呈上升趋势,AG490处理后,系膜细胞中HMGB1mRNA蛋白表达降低,并随时间延长而下调。结论:IFN-γ能够促进小鼠系膜细胞HMGB1表达上调,JAK/STAT信号通路激活可能是其主要机制之一。
Objective：To investigate whether the activation of JAK/STAT signaling pathway by interferon-γ in mesangial cells caused the expression of HMGB1.Methods：Mouse mesangial cell（MMC） selected for the study were randomly divided into control group,IFN-γ stimulation group and IFN-γ＋AG490 group.The cells were collected after 0,2,4,6,8,12 h.Western blot was used to detect the expression of p-JAK1,JAK1,p-JAK2,JAK2,p-STAT1,STAT1 and HMGB1 protein.RT-PCR was used to detect the mRNA of HMGB1.Results：IFN-γ promoted JAK,STAT1 phosphorylation and nuclear translocation of STAT1 in MMC in time-dependent fashion.Specific JAK inhibitor AG490 could reduce IFN-γ-mediated JAK1,JAK2,STAT1 activation,p-JAK1,p-JAK2,p-STAT1 protein expression significantly decreased in the（IFN-γ ＋ AG490） group compared with IFN-γ group in time-dependent manner.IFN-γ up-regulated HMGB1 mRNA and protein expression of MMC cells,the relative level of HMGB1 increased with prolonged stimulation,while AG490 pretreatment inhibited the HMGB1mRNA and protein expression.Conclusion：IFN-γ can upregulate HMGB1 expression in MMC cells,JAK/STAT signaling pathway activation may be one of the main mechanisms.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2012年第10期872-876,共5页
Chinese Journal of Immunology
基金
国家自然科学基金(No.81000301)
河北省自然科学基金(No.C2010000463)
教育部博士点基金(20101323120007)
