目的:观察失荅刺知丸对脑缺血再灌注大鼠海马神经元形态学和Caspase-3、Caspase-9表达的影响。方法:130只健康雄性SD大鼠随机分为假手术组、脑缺血再灌注组(模型组)、失荅刺知丸组(实验组)和金纳多组(阳性对照组),其中后三组根据取材时间点又各分为12 h、24 h和72 h共三个亚组,每组13只。采用线栓法制成大鼠大脑中动脉再灌注模型,透射电镜下观察海马区神经元超微结构变化,免疫组织化学法和Western-blot印迹法检测大鼠海马组织凋亡相关基因Caspase-3、Caspase-9蛋白表达的变化。结果:与假手术组比较,再灌注组大鼠海马神经元出现胞核及线粒体损伤,并且Caspase-3、Caspase-9蛋白的表达明显升高(P〈0.05,P〈0.01);与再灌注组比较,失荅刺知丸组大鼠海马神经元胞核及线粒体损伤明显减轻,而Caspase-3、Caspase-9蛋白的表达明显降低(P〈0.05,P〈0.01)。结论:失荅刺知丸可减轻脑缺血再灌注大鼠海马神经元的损伤,其作用机制可能与抑制Caspase-3、Caspase-9蛋白表达有关。
Objective: To investigate the effects of Shida Lazhi Pill on the morphology and expressions of Caspase-3 and Caspase-9 in hippocampal neurons after cerebral ischemia reperfusion in rats. Methods: 130 Male SD rats were randomly divided into 4 groups,sham-operated group,cerebral ischemia-reperfusion group( reperfusion group),Shida Lazhi Pill intervention group( experiment group) and ginaton intervention group. According to reperfusion time,rats of model were divided into 12,24,72 h subgroups,13 in each group. By using thread-tie method to make middle cerebral artery ischemia-reperfusion model,the changes of hippocampal neurons ultrastructure was observed by transmission electron microscopy. Immunohistochemistry and Western blot were used to measure the expressions of Caspase-3 and Caspase-9. Results: Compared with sham-operated group,nuclear and mitochondrial damage was found in reperfusion group and the expressions of Caspase-3 and Caspase-9 increased obviously in hippocampus( P〈0. 05,P〈0. 01). Compared with the reperfusion group,the damage in nuclear and mitochondria was obviously relieved in experiment group and the expressions of Caspase-3 and Caspase-9 in hippocampus were significantly decreased( P〈0. 05,P〈0. 01). Conclusion: Shida Lazhi Pill can reduce pathological damage of hippocampal neurons after cerebral ischemia and reperfusion in rats and the mechanism is related with inhibiting of Caspase-3 and Caspase-9.