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。
Objective To observe the protective effects of rosmarinic acid（RA） on PC12 cell injured by glutamate,and elucidate whether the NF-κB 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 tetrazolium 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 pretreatment with different concentration of RA（30,60 μmol/L）,it can obviously increase the viability and reduce the apoptosis. Moreover,analysis of western blot indicated that the expression of IkBa in cytoplasm was decreased,the expression of p-IkBa was 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-κB induced by glutamate precondition. Conclusion RA had pro-tective effects on PC12 cell injured by glutamate,which maybe through the inhibition of NF-κB signaling pathway.
出处
《中华老年心脑血管病杂志》
CAS
北大核心
2011年第11期1032-1035,共4页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
