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

Effects of Shida Lazhi Pill on Morphology and Expressions of Caspase-1 and Caspase-9 in Hippocampal Neurons after Cerebral Ischemia Reperfusion in Rats

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.