复方灯盏花滴丸对谷氨酸致原代培养大鼠海马神经元凋亡的保护作用  

Protective effects of Fufangdengzhanhua dripping pill on apoptosis induced by glutamate in cultured primary hippocampal neurons of rats

作　　者：王丽君[1] 宛蕾[1] 

机构地区：[1]贵阳医学院药理学教研室,贵州贵阳550004

出　　处：《中国中药杂志》2010年第5期647-650,共4页China Journal of Chinese Materia Medica

摘　　要：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 explore the protective effects and the inhibited mechanism of Fufangdengzhanhua dripping pill （FDD） on the apoptosis induced by glutamate（Glu） of cultured primary hippocampal neurons of rats. Method： By the seropharmacological method, we obtained the drug-contained sertnn. The primary hippocampal neurons of rat cerebrum were cultured for 10 days, then exposed to 500 umol· L^-1 glutamate acid （Glu）for 20 minutes to build the model. The 5% drug-contained sera which included normal,model, 0. 05 g · kg ^-1 nimodipine（Nim） , 5.00 g · kg^- 1 FDD and 1.25 g · kg^-1 FDD were added to the nutrient solution of cultured neurons. In this study, we observed the following indexes： the viability of cultured primary hippoeampal neurons by MTT assay . the injured cell morphological changes with fluorescence microscope by using Hoechst 33342 ＆ Propicium Iodide（PI） staining, intracellular Ca^2 ＋ concentration and the percentage of apoptosis by flow cytometry. Result ： When tbe hippocampal neurons were exposed to Glu, the cells were seriously damaged ： nuclei were shrunken and cloven and the apoptosis body and the viability of cultured primary hippocampal neurons were decreased dramatically compared with the control. The FDD（5. 00,1. 25 g · kg ^-1 ） and Nim could prevent the above changes Gin-induced. The necrosis rates and the percentage of cellular apoptosis of cultured hippocampal neurons pretreatcd with the serum of containing FDD decreased significantly and the number of surviving cells was increased significantly compared with model. Intracellular Ca^2＋ concentration Glu-induced were increased markedly compared with the control and the FDD（5. （30,1.25 g · kg^-1） could prevent the above changes. Conclusion： FDD has protective effects on tire apoptosis induced by glutamate（Glu） of cultured primary hippocampal neurons of rats, which possibly is related to reducing tire intracellular Ca^2 ＋.

关 键 词：复方灯盏花滴丸 谷氨酸 海马神经元 凋亡 

分 类 号：R285[医药卫生—中药学]