Overexpression of α-synuclein in SH-SY5Y cells in vitro leads to oxidative stress |
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Authors: | ZHANG Yu-xin ZHANG Zi-ming YANG Hui CAI Qing LU Qiang CHEN Biao XU Qun-yuan |
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Affiliation: | 1.North China Coal Medical College, Tangshan 063000, China;2.Capital University of Medical Sciences, Beijing 100054, China |
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Abstract: | 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. |
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