【摘要】 目的研究钙蛋白酶在谷氨酸兴奋性毒性中的表达及钙蛋白酶抑制剂MDL-28170对神经元的保护作用。方法乳大鼠大脑皮质神经元体外培养7 d后用于实验,建立谷氨酸损伤模型,MTT法测定细胞活力,Hoechst33258核染色观察细胞凋亡的形态学改变,Western蛋白印迹法检测钙蛋白酶蛋白水平,免疫荧光染色观察钙蛋白酶在细胞内的表达及定位。结果谷氨酸引起80 kD与76 kD钙蛋白酶蛋白水平明显增加,且分布于凋亡的细胞核内。钙蛋白酶抑制剂MDL-28170明显提高细胞生存率及降低神经元凋亡百分比。结论谷氨酸兴奋性毒性引起钙蛋白酶大量激活,钙蛋白酶抑制剂MDL-28170对谷氨酸引起的神经元损伤具有保护作用。更多还原

【Abstract】 Objective To investigate the expression of Calpain in cultured cortical neurons injured by glutamate and the protective effect of MDL-28170,an calpain inhibitor.Methods The cortical neurons were taken from new born rats and used for experiments 7 days after culture.The neurons were randomly divided into control,glutamate,MDL-28170 + glutamate groups.MTT was used to measure cell viability.Neurons were dyed with Hoechst 33258 and observed under a fluorescence microscope to explore morphology changes.Western blotting was performed to investigate the protein level of calpain.Immunostaining was used to investigate the localization of μ-calpain.Results The protein level of 80 kD and 76 kD Calpain were upregulated after glutamate insult.MDL-28170 increased the cell viability and decreased the apoptotic rate.Conclusions Calpain is severely activated due to the glutamate neurotoxicity.MDL-28170 provides protective effect for the neurons injured by glutamate.更多还原