p38MAPK介导的Fas/FasL凋亡信号通路在大鼠抗GBM肾炎中的作用

摘要

目的:探讨p38MAPK介导的Fas/FasL凋亡信号通路在大鼠抗肾小球基底膜(GBM)肾炎中的作用.方法:1:复制大鼠抗GBM肾炎模型,于实验第2、7、14、21、28天取肾组织行免疫印迹法检测p-p38MAPK、p38MAPK、FasL、Fas、Caspase-3的表达.2:应用p38MAPK阻断剂SB203580处理大鼠抗GBM肾炎模型,于实验第2、7、14天检测24小时尿蛋白、血肌酐及血尿素氮含量;第14天取肾组织,经光镜观察肾组织病理变化;TUNEL法检测肾组织细胞凋亡情况;免疫印迹和免疫组织化学法检测肾组织p-p38MAPK、p38MAPK、FasL、Fas、Caspase-3表达情况.结果:在成功复制大鼠抗GBM肾炎模型基础上,免疫印迹结果显示肾炎模型组大鼠p-p38MAPK、FasL、Fas、Caspase-3蛋白表达各时间点均明显高于正常对照组,而p38MAPK表达与正常对照组相比无明显差异.在成功复制大鼠抗GBM肾炎模型基础上,阻断剂SB203580降低了大鼠抗GBM肾炎生化指标,减轻了肾炎病理反应,抑制了p-p38MAPK、FasL、Fas、Caspase-3的表达,降低了肾组织细胞凋亡数.结论:p38MAPK介导的Fas/FasL凋亡信号通路在大鼠抗GBM肾炎发病机制中发挥重要作用,其阻断剂SB203580减轻了大鼠抗GBM肾炎的病理改变,为临床防治人类新月体性肾炎提供实验理论依据.%Objective:To investigate the effects of Fas/FasL pathway induced by p38MAPK on anti-glomerular basement membrane (GBM) nephritis in rats. Methods;1: Reproduced the anti-GBM nephritis model. The kidney specimens were collected on the 2nd day, the 7th day, the 14th day, the 21st day and the 28th day and the protein expression of p-p38MAPK,p38MAPK, FasL, Fas, Caspase-3 were detected by Western blot. 2: The nephritis rats were injected intraperitoneally with SB 203580 ( an inhibitor of p38MAPK). All rats were observed for 24 h urinary protein, serum creatine, serum urea on the 2nd day, the 7th day and the 14th day. On the 14th day, the kidney specimens were collected to observe the pathological changes ; the numbers of apoptotic cells were detected by TUNEL method; the expression of p-p38MAPK, p38MAPK, FasL, Fas and Caspase-3 were detected by Western blot and immunohistochemical method. Results:Results from immunoblotting showed that p-p38MAPK, FasL, Fas and Caspase-3 expression of the nephritis group were increased obviously compared with the normal control group at each time point , but there is no difference of p38MAPK expression between the two groups. On the anti-GBM nephritis model in rats , SB203580 depressed the biochemical indicators; lessened the pathological changes ; inhibited the expression of p-p38MAPK, FasL, Fas and Caspase-3; reduced the numbers of apoptotic cells. Conclusion:The apoptosis induced by p38MAPK through Fas/FasL pathway play important effects on anti-GBM nephritis in rats. The renal pathological changes of anti -GBM nephritis can be lessened by SB203580, which maybe provide experimental basis for clinical control.