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目的:观察大鼠脑缺血再灌注后脑内脑源性神经营养因子(BDNF)mRNA和蛋白的表达变化以及清热化瘀方对其影响。方法:采用线栓法制作局灶性脑缺血大鼠(MCAO)模型,用原位杂交法和免疫组化法观察脑缺血后3、6、12、24h及3、7d共6个时间点BDNF mRNA及其蛋白的表达变化,以TUNEL法检测神经细胞凋亡。结果:缺血半暗带BDNF mRNA及其蛋白的表达均于缺血再灌注后3h开始明显上升,6h表达继续增强,12h达高峰,3d后表达开始减弱(P<0.05,P<0.01);神经细胞凋亡于缺血再灌注后3h开始明显上升,24h达高峰,3d后表达开始减弱(P<0.05,P<0.01)。清热化瘀方治疗后可以上调BDNF的表达,减轻神经细胞凋亡(P<0.05,P<0.01)。结论:清热化瘀方可促进脑缺血后BDNF的表达,保护神经元。
Objective: To observe the expression of brain-derived neurotrophic factor (BDNF) mRNA and protein in rat brain after cerebral ischemia-reperfusion and the effect of Qingre Huayu Recipe on it. Methods: The model of focal cerebral ischemia (MCAO) was established by thread occlusion method. The in situ hybridization and immunohistochemistry were used to observe the changes of cerebral ischemia at 3, 6, 12, 24 h and 3, 7 d Point BDNF mRNA and protein expression changes, TUNEL assay of nerve cell apoptosis. Results: The expressions of BDNF mRNA and protein in penumbra of ischemic penumbra increased significantly at 3h after ischemia - reperfusion, and continued to increase at 6h. The expression peaked at 12h and began to decrease after 3 days (P <0.05, P <0.01). Neuronal apoptosis increased significantly at 3h after ischemia-reperfusion, peaked at 24h, and began to decrease after 3d (P <0.05, P <0.01). Qingre Huayu Recipe could up-regulate the expression of BDNF and reduce the apoptosis of nerve cells (P <0.05, P <0.01). Conclusion: Qingre Huayu Recipe can promote BDNF expression after cerebral ischemia and protect neurons.