H_2O_2致PC12细胞凋亡与细胞内14-3-3蛋白表达水平下调的关系

目的:探讨用H2O2处理PC12细胞制作PD氧化应激细胞模型的最佳浓度与最适处理时间,以及H2O2致PC12细胞凋亡与14-3-3蛋白表达水平的关系。方法:采用MTT法、荧光分光光度计、流式细胞术及免疫印迹技术分别检测PC12细胞活力、Caspase酶活性、细胞凋亡率以及14-3-3蛋白表达量。结果:当H2O2浓度为100μmol/L时Caspase活性最高;100μmol/LH2O2处理PC12细胞24h时细胞凋亡率最高;PC12细胞的凋亡率与14-3-3蛋白表达的水平呈显著负相关。结论:100μmol/L和24h分别为用H2O2制作PD氧化应激模型的最佳浓度和处理时间,H2O2致PC12细胞的凋亡与细胞内14-3-3蛋白表达水平下调有关。

Apoptosis of PC12 Cells Induced by H2O2 is Correlated with Down-regulated Expression of 14-3-3 Protein

Objective:To explore optimal concentration and time-course of H_2O_2 treatment of PC12 cells for setting up an oxidative stress cell model of PD,and to study relationship between apoptosis of PC12 cells induced by H_2O_2 and expression level of 14-3-3 protein.Methods:MTT assay,fluorescent spectrophotometer,flow cytometric analysis,and western blotting were employed to measure cell viability,caspase activity,apoptotic ratio,and expression level of 14-3-3 protein respectively.Results:Treating PC12 cells with 100 μmol/L for 24 hours yielded the highest caspase activity and apoptotic ratio.A significantly negative correlation was found between expression level of 14-3-3 protein and apoptotic ratio of PC12 cells induced by H_2O_2.Conclusion:Incubation with 100 μmol/L H_2O_2 for 24 hours is the optimal condition for setting up the oxidative stress cell model of PD.The apoptosis of PC12 cells induced by H_2O_2 is correlated with the down-regulated expression of 14-3-3 protein.