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A retinal disorder known as age-related macular degeneration (AMD) can be very damaging, which may result in vision loss. Cordyceps militaris has antioxidant, anti-inflammatory, anticancer activity and immunomodulatory functions. carotenoids were considered potential therapeutic agents for the treatment of age-related macular degeneration. In this study, we evaluated for the first time the protective effect of purified carotenoids (CMCT) extracted from Cordyceps militaris on hydrogen peroxide (H2O2)-induced damage to human retinal pigment epithelial cells (ARPE-19). The pretreatment of ARPE-19 cells with CMCT (1, 2.5μg/ml) for 12h after exposure to H2O2 (400μM) greatly improved cell viability and injury tolerance. and reduced reactive oxygen species production (ROS) and decreased malondialdehyde (MDA) formation. Furthermore, Bax and Caspase-3 showed increased protein expression and less protein expression of Bcl-2 in cellular oxidative stress by flow cytometry analysis but showed the opposite expression by adding CMCT. Besides, cells were treated with H2O2 and then CMCT restored catalase (CAT), superoxide dismutase (SOD) and glutathione (GSH) enzyme activities to normal levels. Our results suggest that CMCT can protect RPE cells from oxidative stress damage by regulating oxidative activity and anti-apoptotic function. It indicates that CMCT has a potential therapeutic role in oxidative stress-directed protection and prevention of AMD.