α-突触核蛋白在体外培养SH-SY5Y细胞内过表达可导致氧应激

目的：观察α-突触核蛋白基因在SH-SY5Y细胞内过表达对细胞的影响。 方法： LipofectAMINE法转染SH-SY5Y细胞，用G418对转基因细胞进行筛选，免疫荧光细胞化学检测α-突触核蛋白表达，氧化应激敏感的荧光素探针DCF-DA对活细胞进行染色后用荧光显微镜和流式细胞仪检测细胞内氧应激状况，还原型谷胱甘肽测定试剂盒通过分光光度计检测细胞内还原型谷胱甘肽水平。 结果： 转染细胞筛选后，细胞呈α-突触核蛋白免疫反应阳性；转α-突触核蛋白基因细胞内存在DCF信号增强和还原型谷胱甘肽水平下降。 结论： α-突触核蛋白过表达可引起细胞内氧应激，表明α-突触核蛋白在帕金森病发病机制中可能起重要作用。

Overexpression of a - synuclein in SH - SY5Y cells in vitro leads to oxidative stress

AIM: To observe the influence of overexpression of α-synuclein on the cultured SH-SY5Y cells. METHODS: The plasmid of α-synuclein-pcDNA3 was transfected into SH-SY5Y cells with LipofectAMINE. The expression of α-synuclein was determined by anti-α-synuclein immunocytochemistry. The intracellular reactive oxygen species was determined with 2', 7'-dichlorofluorescein diacetate (DCF-DA) by using a FACSCAN flow cytometer and fluorescent microscope. The intracellular content of reduced GSH was detected with glutathione assay kit by spectrophotometer. RESULTS: The α-synuclein was expressed in cultured SH-SY5Y cells transfected with the plasmid of α-synuclein-pcDNA3. The DCF loading analysis and the intracellular level of reduced GSH suggested that the transfected cells were under oxidative stress. CONCLUSION: Overexpression and accumulation of α-synuclein in SH-SY5Y cells increase intracellular reactive oxygen species levels, it is suggested therefore that the α-synuclein does play an important role in the pathogenesis of Parkinson's disease.