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D.-N. Hu, R. Rosen, H. Savage, M. Chen, A. Vora, S. A. McCormick; Comparison of Protective Effects of Various Antioxidants Against Hydrogen Peroxide Induced RPE Death in vitro. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5912. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To compare the protective effects of various flavonoids, ascorbic acid, alpha-tocopherol and glutathione against hydrogen peroxide (H2O2) induced retinal pigment epithelial (RPE) cells death in vitro.
Human RPE cells were seeded into 96 cell wells. Fisetin, apigenin, naringin, ascorbic acid, alpha-tocopherol and glutathione were added to the culture medium at various concentrations. Cells cultured with medium without antioxidants were used as the controls. Two hours later, H2O2 was added (0.3 mM). All tests were run in triplicate. After 24 hours, quantity of viable cells was measured using the MTT assay.
All tested antioxidants were not toxic at tested concentration. H2O2 at 0.3 mM decreased the viable cells to 8.1% the controls. Fisetin, ascorbic acid and alpha-tocopherol showed dose-dependent protective effects on H2O2 induced cell death from 10 to300 µM. Viable cells in H2O2-treated RPE cultured with fisetin, apigenin, naringin, ascorbic acid, alpha-tocopherol and glutathione at 100 µM were 56.2%, 7.4%, 9.8%, 17.6%, 24.2% and 10.6% of the controls (without H2O2). The difference of quantity of viable cells between various antioxidants and the control were very significant (P < 0.01) in fisetin, ascorbic acid and alpha-tocopherol treated cells, indicating that these agents protected the RPE cells against the H2O2 cytotoxicity. There was not significant difference in quantity of viable cells between cells cultured with apigenin, naringin and glutathione and the controls, indicating that these agents at 100 µM level could not protect the RPE against H2O2 induced cell death.
In three flavonoids tested, only fisetin (a flavonol) shows protective effects on H2O2 induced RPE death. The other two flavonoids, apigenin (a flavone) and naringin (a flavanone) do not have significant protective effects against oxidative stress. Of three classic antioxidants tested, ascorbic acid and alpha-tocopherol showed protective effects against oxidative stress; glutathione does not, possibly related to the poor passage of glutathione through the cell membrane and enter the cell.
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