Abstract:Objective To investigate the expression of intracellular oxides and antioxidants in human neuroblastoma cells (SH-SY5Y) effected by rotenone.Methods The cells were divided into normal group and rotenone group according different treatment after two days of conventional fetal bovine serum culture.The rotenone group was divided into five subgroups according to the concentration gradient of 25, 50, 100, 200, 500, 1000, 2000 nM (nM: nmol/L).The morphological changes of the cells were observed by light microscope and the survival and growth of the cells were detected by MTT colorimetry.Flow cytometry was used to observe the changes of DCFH-DA in the cells after rotenone intervention, and enzyme-linked immunosorbent assay was used to determine the activity of superoxide dismutase(SOD) and lipid peroxidation.Results It was confirmed that the cell viability of SH-SY5Y was dose-dependent with the intervention concentration of rotenone by MTT assay and light microscopy detection.The concentration of 500 nM rotenone could induce oxidative stress injury without excessive cell death, while the growth of cells almost stopped when 2000 nM rotenone was added into the medium.Flow cytometry and enzyme-linked immunosorbent assay showed that DCFH-DA staining of SH-SY5Y cells treated with 500 nM rotenone increased the level of oxidative stress, decreased the activity of SOD and increased the level of oxide lipid peroxidation products (TBARS).Conclusion Rotenone could regulate down of SOD activity in SH-SY5Y cells, increase the production of lipid peroxidation and increase the level of oxidative stress.The cell viability of SH-SY5Y was positively correlated with the concentration of rotenone.The higher the concentration, the more cell death.
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