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目的:探讨复方灯盏花滴丸对谷氨酸致大鼠原代海马神经元凋亡的保护作用及其作用机制。方法:采用中药血清药理学方法,制备含药血清;实验分为空白对照组、模型组、尼莫地平组(0.05 g.kg-1),复方灯盏花滴丸高、低剂量组(5.00,1.25 g.kg-1),原代培养大鼠乳鼠大脑海马神经元,以谷氨酸(终浓度为500μmol.L-1)作用20 m in复制损伤模型,以5%含药血清干预,MTT法测定细胞存活率,检测细胞内Ca2+浓度,Hoechst33342和PI双染法荧光显微镜观察神经细胞形态学变化和细胞坏死和凋亡百分率,流式细胞仪检测细胞凋亡。结果:谷氨酸对原代培养的海马神经元有损伤作用,荧光显微镜观察到细胞核皱缩、碎裂及凋亡小体,海马神经元的存活率下降,坏死、凋亡明显高于对照组,复方灯盏花滴丸高、低剂量组和尼莫地平组能明显对抗谷氨酸引起的神经细胞形态改变,海马神经元存活率明显提高;复方灯盏花滴丸含药血清组细胞凋亡率较谷氨酸损伤组显著降低,细胞内钙超载明显减轻。结论:复方灯盏花滴丸对谷氨酸致大鼠海马神经元的凋亡具有保护作用,其作用机制可能与其能降低细胞内Ca超载有关。
Objective: To investigate the protective effect of Compound Dengzhanhua Dripping Pill on apoptosis of primary cultured hippocampal neurons induced by glutamate and its mechanism. Methods: Serum containing serum was prepared by the method of traditional Chinese medicine serum pharmacology. The experiment was divided into blank control group, model group, nimodipine group (0.05 g.kg-1), compound Erigeron breviscapus dripping pill high and low dose group (5.00, 1.25 g · kg-1). Primary cultured rat neonatal rat hippocampal neurons were cultured with glutamate (final concentration of 500 μmol·L-1) for 20 min, The cell viability was measured by MTT assay. The intracellular Ca2 + concentration was measured. Hoechst33342 and PI double staining were used to observe the morphological changes of nerve cells and the percentage of cell necrosis and apoptosis. Flow cytometry was used to detect apoptosis. Results: Glutamate could injure the primary cultured hippocampal neurons. The cell nucleus shrinkage, fragmentation and apoptotic bodies were observed by fluorescence microscope. The survival rate of hippocampal neurons decreased, necrosis and apoptosis were significantly higher than those of the control group , The compound Dengzhanhua dripping pills high, low dose group and nimodipine group can obviously antagonize glutamate-induced neuronal morphological changes, hippocampal neuronal survival rate was significantly improved; compound Dengzhanhua droplet containing drug serum group apoptosis rate Compared with glutamate-induced injury group, the intracellular calcium overload was significantly reduced. Conclusion: The compound Dengzhanhua dripping pills can protect rat hippocampal neurons from glutamate-induced apoptosis, and its mechanism may be related to its ability to reduce intracellular Ca overload.