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目的:研究不同纯化工艺制备的健脑益智胶囊对大鼠脑缺血-再灌注损伤的保护作用,探讨其作用机制。方法:120只SD大鼠随机分为假手术组、模型组及健脑益智胶囊原药液组、醇沉组、壳聚组、膜分离组,建立大脑中动脉缺血(MCAO)-再灌注损伤模型,HE染色后观察大鼠缺血侧大脑皮质神经元形态变化,检测血清中丙二醛(MDA)、一氧化氮(NO)、总超氧化物歧化酶(T-SOD)、还原型谷胱甘肽(GSH-Px)的含量,免疫组化法检测脑组织中凋亡相关蛋白BCL-2、BAX的表达。结果:不同纯化工艺制备的健脑益智胶囊均能保护神经元结构,减轻神经元损伤,显著降低血清中MDA、NO水平,提高T-SOD、GSH-Px活性,上调BCL-2蛋白和下调BAX蛋白的表达,其中醇沉组作用更为明显。结论:健脑益智胶囊通过调控BCL-2/BAX相关蛋白的表达,对抗自由基引起的脑组织过氧化损伤,发挥抗凋亡作用。采用醇沉法纯化的健脑益智胶囊对神经保护作用更为显著。
Objective: To study the protective effects of Jiannao Yizhi Capsule prepared by different purification processes on cerebral ischemia-reperfusion injury in rats and to explore its mechanism. Methods: One hundred and twenty SD rats were randomly divided into three groups: sham operation group, model group and Jiannao Yizhi capsule, alcohol precipitation group, chitosan group and membrane separation group, and established middle cerebral artery occlusion (MCAO) The model of perfusion injury was established. Morphological changes of neurons in ischemic cortex of rats were observed by HE staining. Malondialdehyde (MDA), nitric oxide (NO), total superoxide dismutase (T-SOD) The content of glutathione (GSH-Px) and the expression of apoptosis-related proteins BCL-2 and BAX in brain tissues were detected by immunohistochemistry. Results: Jiannao Yizhi Capsule prepared by different purification processes could protect neuron structure, reduce neuronal damage, significantly reduce serum MDA and NO levels, increase T-SOD and GSH-Px activities, up-regulate BCL-2 protein and down-regulate BAX protein expression, which alcohol sedimentation effect is more obvious. Conclusion: Jiannao Yizhi Capsule can play an anti-apoptotic role by regulating the expression of BCL-2 / BAX-related proteins against free radical-induced oxidative injury of brain tissue. The use of Jian Shen Capsule purified alcohol precipitation on neuroprotective effect is more significant.