IFN-γ通过激活JAK_STAT信号转导途径上调MMC内HMGB1表达.docVIP

  IFN-γ通过激活 JAK/STAT 信号转导途径 上调 MMC 内 HMGB1 表达# 封晓娟，张玉军，徐宁，刘淑霞** 5 10 15 20 25 30 35 （河北医科大学病理教研室，石家庄 050017） 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（mouse mesangial cell, MMC），无血清培养基 同步化后分为正常对照组、IFN-γ(500 IU/ml)刺激组和 INF-γ+AG490 ( INF-γ500 IU/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 1.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；2. AG490 能够降低 IFN-γ 介导的 JAK1，JAK2，STAT1 的活化，并呈时间依赖性。3. 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。 关键词：狼疮性肾炎；IFN-γ；JAK/STAT；HMGB1 中图分类号：R363，R593 Interferon-γ Induces the expression of HMGB1 in mouse mesangial cell by activating JAK / STAT signaling pathway FENG Xiaojuan, ZHANG Yujun, XU Ning, LIU Shuxia (Department of Pathology, Hebei Medical University, ShiJiaZhuang 050017) Abstract: Objective To detect the expression of p-JAK1, JAK1, p-JAK2, JAK2, p-STAT1, STAT1 and HMGB1 in order to investigate whether the activation of JAK / STAT signaling pathway by interferon-γ in mesangial cells caused the expression of HMGB1. Methods 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 bloting 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 1. IFN-γ promoted JAK, STAT1 phosphorylation and nuclear translocation of STAT1 in MMC in time-dependent fashion. 2. 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. 3. IFN-γ up-regulated HMGB1 mRNA and protein expression of MMC cells, the re