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目的 研究α synuclein的纤维源性片段NAC对PC12细胞内蛋白聚集、线粒体功能及活性氧水平的影响。 方法 采用神经生长因子将PC12细胞诱导分化成多巴胺能细胞 ,后经不同浓度的NAC(0、0 1、1 0、10 0、2 5 0、5 0 0 μmol/L)处理 4 8h ,MTT法测定线粒体氧化还原酶的活力。用硫磺素S染色观察细胞内蛋白聚集情况 ,JC 1检测线粒体膜电位 ,DCFH DA检测细胞内活性氧水平。 结果 经NAC处理 4 8h后 ,与对照组比较 ,PC12细胞线粒体活力在高浓度 (≥ 2 5 μmol/L)NAC作用时明显下降 6 4 1% (P <0 0 5 )。NAC(2 5 μmol/L)可诱导出现蛋白聚集 ,JC 1荧光染色可检测到线粒体膜电位明显去极化 ,DCFH DA荧光染色及流式细胞仪检测显示PC12细胞内活性氧增加。 6 0 6 % 结论 NAC可引起PC12细胞内蛋白聚集 ,进而线粒体功能障碍及氧化应激增强 ,最终导致细胞死亡。
Objective To investigate the effects of NAC, an α-synuclein-derived fiber, on protein accumulation, mitochondrial function and reactive oxygen species in PC12 cells. Methods PC12 cells were induced to differentiate into dopaminergic cells by using nerve growth factor (NGF), and then treated with various concentrations of NAC (0, 0, 110, 105, 500 μmol / L) for 48 h. MTT assay Mitochondrial oxidoreductase activity was measured. Thioflavin S staining was used to observe intracellular protein aggregation, JC 1 detection of mitochondrial membrane potential, DCFH DA detection of intracellular reactive oxygen species. Results Compared with the control group, the mitochondrial viability of PC12 cells significantly decreased by 64.1% (P <0.05) at NAC concentrations ≥ 25 μmol / L after 48 h treatment with NAC. NAC (2 5 μmol / L) could induce protein aggregation, and JC 1 fluorescence staining could detect significant depolarization of mitochondrial membrane potential. Fluorescence staining with DCFH DA and flow cytometry showed that reactive oxygen species increased in PC12 cells. 6 0 6% Conclusion NAC can cause protein accumulation in PC12 cells, which in turn leads to mitochondrial dysfunction and increased oxidative stress leading to cell death.