The effects of crocetin on oxidative stress induced ARPE-19 cells by H₂O₂

The anti-apoptotic and antioxidant effects of crocetin was aimed to investigate on the oxidative damage model of ARPE-19 cells. The oxidative damage in ARPE cells was developed by H₂O₂ treatment at 800 μM. Different doses of crocetin (1-80 μM) were applied for 24 h, and the effects on viability were evaluated to find out the optimum drug dose. At first, three effective doses of crocetin (10, 20, 40 μM) on cell viability were selected for further analyses. The levels of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione (GSH) were determined, and the expression of pro-apoptotic Bax gene and anti-apoptotic Bcl-2 gene were evaluated. The most effective crocetin dose on cell viability was found to be 10 μM. After the H₂O₂ treatment, SOD and GSH were decreased and MDA were increased significantly (p = 0.011, 0.037, 0.018, respectively). Following the crocetin treatment at 10 μM, SOD and GSH activities were improved compared to the no drug group; and MDA level was declined remarkably (p = 0.022, 0.019, 0.029, respectively). The Bcl-2 level was significantly decreased (p < 0.01), while the Bax1 and Nrf2 expression and ROS level was increased significantly in the damage model group (p < 0.01). After the drug treatment, the Bax1 and Nrf2 expression level were decreased in all groups (p < 0.01). The increase in Bcl-2 expression was significant in crocetin 40 μM (p < 0.05) and the decrease in ROS level were significant in 20 μM and 40 μM doses of crocetin (p < 0.05). It has been shown that crocetin might be used as an antioxidant and anti-apoptotic agent on the hindering the effect of the oxidative damage. Following the development of the oxidative stress in the cells, crocetin reversed the damage signals. By the in vitro tests, it was shown that crocetin might be considered as an effective molecule to be used in the AMD treatment.