摘要
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。
Objective：To explore optimal concentration and time-course of H2O2 treatment of PC12 cells for setting up an oxidative stress ceil model of PD,and to study relationship between apoptosis of PC12 ceils induced by H2O2 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, apoptofic 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 H2O2. Conclusion：Incubation with 100 μmol/L H2O2 for 24 hours is the optimal condition for setting up the oxidative stress cell model of PD. The apoptosis of PC12 cells induced by H2O2 is correlated with the down-regulated expression of 14-3-3 protein.
出处
《神经损伤与功能重建》
2008年第1期19-21,37,共4页
Neural Injury and Functional Reconstruction
