Protective Effect of Diosgenin against H2O2-Induced Oxidative Stress on H9C2 Cells

Abstract

Diosgenin is an important compound in pharmaceutical industry. It has various effects such as hypocholesterolemic action or antioxidant activity in HIV infected patients. Biological oxidation pathways are involved in causing or aggravating heart disease. This study investigated the potential protective effect of diosgenin on cell viability and antioxidant defenses of cultured H9C2 cells submitted to oxidative stress induced by H2O2. Viability of cells exposed to H2O2 was detected by MTT assay. The generation of ROS and hydrogen peroxide release after H2O2 were detected using the fluorescent probe H2DCF-DA. The lipid peroxidation product i.e. MDA formation was estimated by assessing the levels of thio-barbituric acid reactive substances (TBARS) using spectrophotometry. SOD activity was assayed with NWLSS (TM) Superoxide Dismutase (SOD) activity assay kit. Pretreatment of cells with 3-25 µM of diosgenin for 24 h before applying H2O2 completely prevented cell damage and significantly enhanced viability of H9C2 cells. Increased ROS induced by H2O2 was dose dependently prevented when cells were pretreated for 24 h with diosgenin. The level of the lipid peroxidation was significantly higher in H9C2 cells exposed to H2O2 as compared to the control and cells pretreated with diosgenin. SOD activity in cells treated with diosgenin significantly decreased compared with cells exposed to H2O2. These results show that treatment of H9C2 cells with diosgenin (3-25 µM) confers a significant protection against oxidative stress.