Overexpression of ?-synuclein in SH-SY5Y cells in vitro leads to oxidative stress / 中国病理生理杂志

ABSTRACT

AIM: To observe the influence of overexpression of ?-synuclein on the cultured SH-SY5Y cells. METHODS: The plasmid of ?-synuclein-pcDNA_3 was transfected into SH-SY5Y cells with LipofectAMINE. The expression of ?-synuclein was determined by anti-?-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 ?-synuclein was expressed in cultured SH-SY5Y cells transfected with the plasmid of ?-synuclein-pcDNA_3. 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 ?-synuclein in SH-SY5Y cells increase intracellular reactive oxygen species levels, it is suggested therefore that the ?-synuclein does play an important role in the pathogenesis of Parkinson's disease.