Fas/FasL途径在海水诱导大鼠肺组织细胞凋亡中的作用

Effect of Fas/FasL-mediated pathway on seawater-induced cell apoptosis in lung tissues of rats

目的通过观察和验证Fas/FasL信号途径对海水诱导大鼠肺组织细胞凋亡的影响,研究海水淹溺型肺损伤的致病机制。方法 24只大鼠随机分为正常对照组、2 h海水组和4 h海水组。2个海水处理组大鼠气管内注入4 ml/kg海水,分别作用2 h和4 h,测肺组织湿重/干重(W/D)比,采用HE染色观察肺组织病理形态改变,流式细胞仪检测肺组织细胞凋亡,蛋白质免疫印迹法(Western blotting)分析Fas、FasL及活化caspase-3的蛋白表达。结果与正常对照组相比,海水可导致典型的肺组织损伤,海水作用下肺组织细胞凋亡明显增加(P<0.01),Fas、FasL和活化caspase-3蛋白表达也显著上调(P<0.01)。结论海水可导致大鼠淹溺型肺损伤和肺组织细胞凋亡,Fas/FasL介导的死亡受体信号途径参与了凋亡的调控。

Objective To observe and verify the influence of Fas/FasL signaling pathway on seawater-induced lung tissue cell apoptosis of rat,and to explore the mechanism of seawater drowning-induced acute lung injury（SWD-ALI）.Methods Twenty-four rats were randomly divided into three groups：normal control group,2 h seawater group and 4 h seawater group.Seawater（4 ml/kg） was instilled into both lungs of the rats and then the rats were killed at 2 h and 4 h in seawater groups,respectively.Lung wet/dry weight was calculated.The morphological changes of lung tissues were observed by HE staining.The cell apoptosis in rat lung tissues was measured by flow cytometry（FCM）.The protein expression of Fas,FasL and caspase-3 was analyzed by Western blotting.Results The typical lung tissue injury was successfully induced in Seawater groups.Compared with control group,lung tissue cell apoptosis increased significantly in seawater groups（P0.01）,and the protein expression of Fas,FasL and caspase-3 was obviously up-regulated in seawater groups（P0.01）.Conclusion Seawater could lead to the rat lung injury and lung tissue cell apoptosis,and the Fas/FasL-mediated death signaling pathway is involved in the regulation of apoptosis.