Protective effects of paeoniflorin against hydrogen peroxide-induced cell injury in SH-SY5Y nerve cells / 中草药

ABSTRACT

Objective: To investigate the neuroprotective effects of paeoniflorin (PF) against cell death induced by hydrogen peroxide (H2O2) in human neuroblastoma cells and its mechanisms. Methods: H2O2 was used to induce SH-SY5Y cell damages, and the cell survival rate was detected by CCK-8 assay; the cell morphologic changes were observed by inverted optical microscope; the apoptosis was tested using Hoechst 33258 staining; flow cytometer (FCM) and propidium iodide staining were used to analyze the apoptosis and cell cycle alteration; reactive oxygen species (ROS) production was determined by 2', 7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescence; lactic dehydrogenase (LDH) release was detected by reaction of diaphorase and INT; 8-OHdG production was determined by enzyme-linked immunosorbent assay (ELISA); caspase-3 activity was determined by caspase-3 catalyzing the specific substrate. Results: Compared with control group, after the treatment with H2O2 (200 μmol/L) for 24 h, the viability and proliferation index of CH-SY5Y cells were significantly decreased (P < 0.01, 0.05), the apoptosis rate and content of 8-OHdG were increased (P < 0.01), the LDH resease and ROS production were increased (P < 0.01); the activity of caspase 3 was increased (P < 0.01). Compared with H2O2 injury group, PF (20-40 μmol/L) significantly ameliorated the changes in SH-SY5Y cells induced by H2O2 in concentration-related manner (P < 0.05). PF (10 μmol/L) did not significantly change the above indexes except the cell viability, ROS, and caspase-3 activity induced by H2O2 (P < 0.05). Conclusion: PF has the significant protective effect against the H2O2-induced cell injury, which may be related to eliminatinging ROS, alleviating DNA oxidative damage, regulation of cell cycle, and inhibition of apoptosis of caspase pathway activation.