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目的探讨芪棱汤对大鼠局灶性脑缺血再灌注不同时间点caspase-9表达的影响及其对局灶性脑缺血再灌注损伤的保护作用。方法采用改良线栓法制备大鼠大脑中动脉阻塞致短暂局灶性脑缺血模型,于各灌注时间点拔出线栓使血流再通,形成大鼠脑缺血再灌注模型。大鼠随机分为假手术组、生理盐水组、芪棱汤去养阴药组、芪棱汤组,于缺血2h再灌注2、4、6、12、24、48h进行神经功能评分,用免疫组织化学法检测各组梗死边缘区caspase-9表达的动态变化。结果各组大鼠神经功能评分差异均有显著性,芪棱汤组明显优于其他各组。随着再灌注时间的延长,caspase-9阳性细胞计数的表达不断增加,至再灌注24h时到达高峰,再灌注48h时阳性细胞计数表达开始减少。再灌注各时点各组阳性细胞计数显示,芪棱汤去养阴药组和芪棱汤组的阳性细胞数均少于对照组;再灌注4h时芪棱汤组与芪棱汤去养阴药组阳性细胞计数差异无显著性;再灌注6、12、24、48h时芪棱汤组阳性细胞数少于芪棱汤去养阴药组。结论芪棱汤与芪棱汤去养阴药可能通过抑制caspase-9的释放和激活而减少脑缺血再灌注损伤,且芪棱汤疗效优于芪棱汤去养阴药。
Objective To investigate the effect of Qileng Decoction on the expression of caspase-9 at different time points after focal cerebral ischemia and reperfusion in rats and its protective effect on focal cerebral ischemia-reperfusion injury. Methods The model of transient focal cerebral ischemia induced by occlusion of middle cerebral artery in rats was prepared by modified suture embolization method. The line was punctured to recanalize the blood flow at each time of perfusion to form a model of cerebral ischemia and reperfusion in rats. Rats were randomly divided into sham-operated group, normal saline group, Qileng Decoction, and Qileng Decoction group. Neurological function was scored at 2, 4, 6, 12, 24, and 48 hours after ischemia for 2 hours. Immunohistochemistry was used to detect the dynamic changes of caspase-9 expression in the infarct margins of each group. Results The scores of neurological function in each group were significantly different. The Qileng Decoction group was significantly better than other groups. With the increase of reperfusion time, the expression of caspase-9 positive cell count increased continuously, reached the peak at 24 hours after reperfusion, and the expression of positive cell count began to decrease at 48 hours after reperfusion. The number of positive cells in each group at each time point of reperfusion showed that the number of positive cells in the Qileng Decoction to Yangyin Decoction group and Qileng Decoction group were all less than those in the control group; when reperfusion was performed for 4 hours, Qileng Decoction and Qileng Decoction went to nourish Yin. There was no significant difference in the number of positive cell counts in the drug group; the number of positive cells in the Qileng Decoction group was less than that of the Qileng Decoction and Yin Yang Decoction group at 6, 12, 24, and 48 hours after reperfusion. Conclusion Cangling Decoction and Cangling Decoction can decrease the cerebral ischemia-reperfusion injury by inhibiting the release and activation of caspase-9, and the curative effect of Qileng Decoction is better than that of Qilang Decoction and Yin Yang Decoction.