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Oxidative stress and free radical production have been implicated in Alzheimer's disease, where low levels of the antioxidant vitamin C (ascorbate) have been shown to be associated with the disease. In this study, neuroblastoma SH-SY5Y cells were treated with hydrogen peroxide in the presence of ascorbate in order to elucidate the mechanism(s) of protection against oxidative stress afforded by ascorbate. Protein oxidation, glutathione levels, cell viability and the effects on the proteome and its oxidized counterpart were monitored. SH-SY5Y cells treated with ascorbate prior to co-incubation with peroxide showed increased viability in comparison to cells treated with peroxide alone. This dual treatment also caused an increase in protein carbonyl content and a decrease in glutathione levels within the cells. Proteins, extracted from SH-SY5Y cells that were treated with either ascorbate or peroxide alone or with ascorbate prior to peroxide, were separated by two-dimensional gel electrophoresis and analyzed for oxidation. Co-incubation for 24 hours decreased the number of oxidised proteins (e.g. acyl CoA oxidase 3) and induced brain derived neurotrophic factor (BDNF) expression. Enhanced expression of BDNF may contribute to the protective effects of ascorbate against oxidative stress in neuronal cells.

Original publication

DOI

10.1002/pmic.200300924

Type

Journal article

Journal

Proteomics

Publication Date

02/2005

Volume

5

Pages

534 - 540

Keywords

Antioxidants, Ascorbic Acid, Brain-Derived Neurotrophic Factor, Cell Line, Tumor, Cell Survival, Dose-Response Relationship, Drug, Electrophoresis, Gel, Two-Dimensional, Enzyme-Linked Immunosorbent Assay, Glutathione, Humans, Hydrogen Peroxide, Neuroblastoma, Oxidants, Oxidation-Reduction, Oxidative Stress, Proteins, Proteome