小续命汤对急性脑缺血再灌注线粒体相关蛋白Hsp60、Mitofilin表达的影响

Xiaoxuming Tang Has Effect on Expression of Mitochondria-related Protein Hsp60 and Mitofilin in Acute Cerebral Ischemia-reperfusion

目的:观察小续命汤对大鼠急性脑缺血再灌注损伤的神经保护作用及对线粒体相关蛋白表达的影响。方法:60只雄性SPF级SD大鼠随机分为假手术组、模型组、小续命汤组、环孢素A组、溶媒组5组,每组12只。采用大脑中动脉线栓法建立脑缺血再灌注损伤模型,观察各组大鼠的一般情况、神经功能缺损,采用Nissl染色观察神经细胞损伤,Western blot和免疫组织化学法检测脑缺血再灌注后缺血半暗带皮层线粒体热休克蛋白60 (Heat shock protein 60,Hsp60)和mitofilin蛋白的表达。结果:与假手术组比较,模型组出现明显神经功能缺损,细胞发生缺血再灌注损伤,缺血半暗带皮层Hsp60、mitofilin蛋白的表达上调(P <0.05)。与模型组比较,小续命汤组、环孢素A组可显著改善大鼠神经功能缺损评分,减轻神经细胞的缺血再灌注损伤,并显著上调脑缺血半暗带皮层Hsp60、mitofilin蛋白的表达,差异均有统计学意义(P <0.05)。结论:小续命汤可改善大鼠神经功能缺损评分,减轻神经细胞的缺血再灌注损伤,上调脑缺血半暗带皮层Hsp60、Mitofilin蛋白的表达,从而发挥脑保护的作用。

Objective： To observe the neuroprotective effect of Xiaoxuming tang on rats with acute cerebral ischemia-reperfusion injury and its effect on the expression of mitochondria-related protein. Methods： Divided 60 male SD rats in SPF grade into 5 groups randomly including the sham operation group,the model group,the Xiaoxuming tang group,the cyclosporine group A and the solvent group,12 rats in each group. Established the model of cerebral ischemia-reperfusion injury by using the method of suture embolization in middle cerebral artery, and observed the general condition and neurologic impairment（NIHSS） of rats in each group. Observed the injury of nerve cells via Nissl staining,and detected the expression of heat shock protein 60（Hsp60） and mitofilin protein in mitochondria of ischemic penumbra cortexafter cerebral ischemia-reperfusion via Western blot and immunohistochemistry. Results： Comparing with the sham operation group,the model group appeared evident neurologic impairment and ischemia-reperfusion injury in cells,which also induced the up-regulation of the expression of Hsp60 and mitofilin protein in ischemic penumbra cortex（P〈0.05）. Comparing with the model group, the scores of NIHSS were improved, ischemia-reperfusion injury of nerve cells was alleviated, and the expression of Hsp60 and mitofilin protein in ischemic penumbra cortex was up-regulated in the Xiaoxuming tang group and the cyclosporine group A,difference being significant（P〈0.05）. Conclusion： Xiaoxuming tang can improve the scores of NIHSS in rats, alleviate ischemia-reperfusion injury of nerve cells and up-regulate the expression of Hsp60 and mitofilin protein in ischemic penumbracortex,thus to take effect in cerebral protection.