[Overexpression of alpha-synuclein in SH-SY5Y cells partially protected against oxidative stress induced by rotenone].
Both genetic and environmental factors are involved in the pathogenesis of Parkinsonos disease (PD). Epidemiological studies showed that environmental factors shared with the common mechanisms of resulting in alpha-synuclein aggregation by inhibiting complex I of mitochondria and leading to oxidative stress. To investigate the relationship between alpha-synuclein and oxidative stress, we used human dopaminergic SH-SY5Y cells transfected with alpha-synuclein-enhanced green fluorescent protein (EGFP). alpha-synuclein gene expression was determined by immunocytochemistry and real-time quantitative PCR. Both SH-SY5Y and alpha-synuclein overexpressed SH-SY5Y (SH-SY5Y/Syn) cells were treated with various concentrations of rotenone for different time. Cell viability and oxidative stress were detected by MTT assay and DCF assay. Superoxide dismutase (SOD) activity was assessed with xanthine peroxidase method. Cell apoptosis was detected with flow cytometry. Results showed that alpha-synuclein gene was constantly overexpressed in SH-SY5Y/Syn cells. After treatment with rotenone, both cell viability and complex I activity in these cells were reduced in a concentration-dependent manner. Oxidative stress was also found in these cells. Compared with SH-SY5Y cells, SOD activity in SH-SY5Y/Syn cells was increased distinctly (P<0.05) and alpha-synuclein significantly attenuated rotenone-induced cell apoptosis. These results suggest that the alpha-synuclein overexpression in SH-SY5Y cells has a tendency to partially resist oxidative stress induced by rotenone and this response may assist cell survival.
Liu
Yan-Ying
YY
Beijing Institute for Neuroscience, Beijing Center for Neural Regeneration and Repair, Key Laboratory for Neurodegenerative Disease of the Ministry of Education of China, Capital Medical University, Beijing 100069, China.
Zhao
Huan-Ying
HY
Zhao
Chun-Li
CL
Duan
Chun-Li
CL
Lu
Ling-Ling
LL
Yang
Hui
H
D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D019610:Cytoprotection; D004305:Dose-Response Relationship, Drug; D042967:Electron Transport Complex I / Q000378:metabolism; D006801:Humans; D018384:Oxidative Stress; D012402:Rotenone / Q000633:toxicity*; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism; D051844:alpha-Synuclein / Q000235:genetics / Q000502:physiology*
D051844:alpha-Synuclein; D012402:Rotenone; D013481:Superoxides; D013482:Superoxide Dismutase; D042967:Electron Transport Complex I