Establishment of an oxidative damage model induced by hydrogen peroxid in TM3 cells

ObjectiveTo establish an oxidative damage model with hydrogen peroxide(H₂O₂)in mouse Leydig(TM3)cells for studying possible mechanism of decreased reproductive function in obese males.MethodsAfter 4 hours' treatment with different concentration of H₂O₂, the cell viability was determined with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT)method;the testosterone level was determined with enzyme-linked immunosorbent assay(ELISA);the contents of reactive oxygen species(ROS),total antioxidant capacity(TAOC)and activities of superoxide dismutase(SOD) and catalase(CAT)were detected with spectrophotometry.ResultsThe viability of TM3 cells (86.7±4.3% and 46.3±2.0%)of 300 and 600 μmol/L H₂O₂ exposure groups decreased significantly compared to that of the control group(100±1.7%)(all P<0.05).The testosterone level(188.93±7.06,179.23±6.66,and 131.59±11.26 pg/mL)of 150,300,600 μmol/L H₂O₂ exposure groups were significantly lower than that of the control group(250.09±26.70 pg/mL)(all P<0.05).The activity of SOD(2.42±0.17,2.69±0.30,and 0.44±0.41 U/mg protein)and CAT(56.76±12.17,23.45±2.17,and 24.55±18.05 U/mg protein)of 150,300,and 600 μmol/L H₂O₂ exposure groups decreased significantly compared to those of the control group(all P<0.05).ConclusionThe TM3 cell model of oxidative injury was established successfully with the administration of H₂O₂.