Protective effect of crocetin on hydrogen peroxide-induced cell injury

Objective To investigate the protective effect of crocetin in H2O2 induced rat pheochromocytoma line PC12,and explore the mechanisms on the injury.Methods The cell viability was detected by Cell Counting Kit-8(CCK-8) assay.Mitochondrial membrane potential(MMP) was stained by the Rhodamine123 and determined by flow cytometry.Reactive oxygen species(ROS) was observed by using DCF-DA staining and photofluorography.Phosphorylation of ERK1/2 was measured by Western blot.Results At the concentrations from 0 to 400 μmol/L for 12 h,the cell viability are(100±4.1)% 、(102±1.9)% 、(89±11.2)% 、(52±2.6)%、(42±1.6)%、(8±0.4)%,respectively.H2O2 dose-dependently inhibited cell viability.Exposure of the cells to different doses of crocetin(0～10 μmol/L) for 24 h followed by subsequent exposure to a single dose of H2O2(200 μmol/L),the cell viability increased from(45.12±3.15)% to(51.88±4.24)%、(65.14±8.19)%、(57.66±5.58)%、(53.61±4.57)%.Preincubation of cells with crocetin(1 μmol/L,5 μmol/L) 24 h prior to H2O2 exposure attenuated the decrease of MMP,scavenged ROS formation and activated ERK1/2 phosphorylation.Conclusion The crocetin holds potential for protective effects against H2O2-induced injury.