Protective Effect of Orychophragmuspine I Against Oxidative Damage in HepG2 Cells Induced by Hydrogen Peroxide
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Abstract:
Orychophragmus violaceus seed extract was isolated and purified by silica gel chromatography, and the structures were identified by spectral analysis and chemical methods. A new compound was identified for the first time and named as Orychophragmuspine I. The protective effect and mechanism of Orychophragmuspine I on hydrogen peroxide (H2O2)-induced oxidative damage in HepG2 cells was investigated by establishing a model of H2O2-induced cell damage. Before hydrogen peroxide stimulation, HepG2 cells were pretreated with different concentrations of Orychophragmuspine I for 12 h, and then cells were treated with 400 μmol/L H2O2 for two hours. The survival rate of cells was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Lactate dehydrogenase (LDH) activity in the medium, cellular malondialdehyde (MDA) content, and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) were measured using a microplate method, and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and heme oxygenase-1 (HO-1) protein expression was measured by western blot. The results showed that, compared with the model group, 12 h of Orychophragmuspine I treatment (53.5, 107, and 214 μmol/L) increased the cell survival rate (p<0.01), significantly reduced the release of LDH to the extracellular fluid (p<0.01), decreased MDA content in cells (p<0.05, p<0.01), and increased the intracellular activities of SOD and GSH-PX (p<0.05, p<0.01) and the level of Nrf2 protein. Therefore, Orychophragmuspine I has a protective effect against H2O2-induced oxidative damage in HepG2 cells.
