目的 探讨p38/Fas/FasL对大鼠心肌缺血再灌注损伤诱导细胞凋亡的调控作用.方法 SD大鼠20只随机分成2组(假手术组、模型组),每组10只.结扎冠状动脉左前降支,30 min后剪断结扎线制作心肌缺血再灌注模型.对心肌组织进行苏木精-伊红(H-E)染色,观察心肌组织病理变化;免疫印迹(Western Blotting)检测Fas、Fas配体(FasL)、p38促分裂素原活化蛋白激酶(p38 MAPK)以及磷酸化p38 MAPK(p-p38 MAPK)在缺血再灌注心肌组织中的表达;TdT介导的dUTP缺口末端标记技术(TUNEL)法检测心肌细胞凋亡模型.结果 H-E染色结果显示,模型组心肌纤维变性;Western Blotting结果显示模型组大鼠心肌组织中Fas、FasL、p38 MAPK及p-p38 MAPK蛋白的表达明显增高;TUNEL结果显示模型组的心肌组织中细胞凋亡明显增多.结论 心肌缺血再灌注时Fas、FasL、p38 MAPK和p-p38 MAPK表达明显增多,且与细胞凋亡率呈正相关,提示心肌缺血再灌注损伤诱导的心肌细胞凋亡可能是通过激活Fas/FasL/p38 MAPK途径实现的.%Objective To investigate the effects of p38/Fas/FasL on myocardial apoptosis in rats with ischemia/ reperfusion injury. Methods Twenty male Sprague - Dawley rats were selected and randomly divided into two groups (n = 10 each) : ① Sham - operated group ; ② ischemia/reperfusion (I/R) group. The myocardial ischemia/reperfusion injury model was established by ligating left anterior descending coronary artery for 30 min followed by 2 h reperfusion. The cardiac muscle tissue was stained by hematoxylin and eosin (H - E) to observe its pathological changes. Western blotting was used to detect the expression of Fas /FasL/p38 MAPK/p - p38 MAPK. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was used to detect the cardiomyocyte apoptosis . Results H - E staining showed that cardiac muscle cells were degenerated in I/R group. Western blotting showed that the expressions of Fas/FasL/p38 MAPK/p -p38 MAPK were significantly higher in ischemic area in the IR group compared to sham -operated group (P <0.01). TUNEL result showed that apoptotic cells increased in I/R group compared to sham - operated group (P < 0.01). Conclusion Fas/FasL/p38 MAPK/p - p38 MAPK are expressed aboundantly following myocardial ischemia / reperfusion and the expressions are positively correlated with apoptosis rate . The results suggest that the mechanism of myocardial apoptosis induced by myocardial ischemia /reperfusion injury may be associated with the activation of Fas / FasL/p38MAPK signal pathway.
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