【摘要】 目的观察迷迭香酸(RA)对抗谷氨酸诱导的PC12细胞损伤发挥神经保护作用,探讨其机制是否与NF-κB信号通路相关。方法将培养细胞分为对照组、损伤组(16 mmol/L谷氨酸)、RA1组(30μmol/L RA+16 mmol/L谷氨酸)、RA2组(60μmol/L RA+16 mmol/L谷氨酸)。采用MTT法检测细胞活力;Hoechst33258荧光染色观察细胞核形态的改变,碘化丙啶染色流式细胞仪检测细胞凋亡;Western blot检测NF-κB信号通路相关蛋白表达水平的变化。结果不同浓度谷氨酸(0、4、8、12、16、20 mmol/L)作用PC12细胞24 h后,细胞存活率呈剂量依赖性下降。与损伤组比较,RA1组和RA2组细胞存活率明显升高(P<0.05)。16 mmol/L谷氨酸作用PC12细胞1 h后,细胞质中IκBα蛋白和NF-κB p65蛋白表达明显减少,p-IκBα蛋白表达明显增加,NF-κB p65蛋白在细胞核中表达明显增加,2 h达到高峰;给予30、60μmol/L RA预处理1 h后,能抑制NF-κB p65蛋白活化进入细胞核。结论 RA通过抑制NF-κB信号通路的活化,对谷氨酸诱导损伤的PC12细胞有保护作用。更多还原

【Abstract】 Objective To observe the protective effects of rosmarinic acid(RA) on PC12 cell injured by glutamate,and elucidate whether the NF-kB signal pathway was involved in this mechanism. Methods The cultured cells were divided into four groups:the control group,the injury group, the RA1 group and the RA2 group.Cell viability was evaluated using the methyl thiazolyl tetrazo-lium assay.Apoptosis was assessed using Hoechst 33258 staining and flow cytometry analysis. NF-κB signaling pathway protein expression levels were detected by Western blot analysis.Results After PC12 cell was incubated with different concentration glutamate(0、4、8、12、16、20 mmol/ L) for 24 hours,the cell survival was reduced in a dose-dependent manner.After an hour pretreat-ment with different concentration of RA(30、60μmol/L),it can obviously increase the viability and reduce the apoptosis.Moreover,analysis of western blot indicated that the expression of IκBαin cytoplasm was decreased,the expression of p-IκBαwas increased,and the expression of NF-κB p65 in cytoplasm was decreased but in the nucelus was increased after treatment of 16 mmol/L glutamate for an hour,and the peak time of these changes were in the two hours.In addition,RA can inhibited the activation of NF-kB induced by glutamate precondition.Conclusion RA had protective effects on PC12 cell injured by glutamate,which maybe through the inhibition of NF-kB signaling pathway.更多还原