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In vitro protection against hydrogen peroxide-induced oxidative stress and cell death in ARPE-19 cells by Curcumin
Curcumin, the main active phenolic compound present in turmeric, possesses a wide range of pharmacological properties including antioxidant activities . Although the exact etiology of age-related macular degeneration (AMD), the leading cause of blindness in the elderly, is not known, the retinal pigment epithelium (RPE) is considered a primary target via oxidative cell damage . The purpose of this study is to determine if curcumin protects RPE cells against H2O2-induced oxidative stress and cell death. Cultured RPE cells (ARPE-19) were pretreated with curcumin (25–100µM) for 1h before washing and incubation with H2O2 (100–600µM) for 24h. The percentage of viable cells was determined using trypan blue dye exclusion. Nuclear chromatin condensation, a hallmark of apoptosis, was determined using Hoechst 33342 staining and then visualized by fluorescence microscopy. The results demonstrated that treatment of ARPE-19 with H2O2 resulted in dose-dependent cell death and an increase in the number of cells containing condensed nuclei in a time-dependent manner. Pretreating cells with 100µM curcumin for 1h prior exposure to 600µM H2O2 notably attenuated cell death and decreased the percentage of cells containing condensed nuclei. These results indicate that in vitro, curcumin provides substantial protection against H2O2-induced cell death and nuclear condensation in ARPE-19 cells and may therefore, be useful as a potential agent in the treatment of disorders associated with oxidative stress-induced cell damage such as AMD.
Acknowledgements: This work was supported partially by research grants from Suranaree University of Technology and Mahidol University.
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