| Objective The aim of the present study was to investigate whether the total flavonoids(TFs)from the leaves of Carya cathayensis Sarg,(LCC)and its five monomeric flavonoids could prevent hydrogen peroxide induced oxidative injury and apoptosis in rat aortic endothelial cell line(SVARECs),and to evaluate their protective effect and possible mechanism.Methods SVARECs were cultured in vitro and the growth curves were draw.Different concentration of hydrogen peroxide(H2O2)were stimulated the cells,and the cell viability were measured by the method of MTS at different time.The non-toxic concentrations of flavonoids were ensured by MTS assay.Cell morphologies,cytotoxicity and apoptosis were assessed by light microscopy,MTS assay,Annexin V-FITC and propidium iodide double staining respectively.MDA,SOD,LDH and GSH-PX levels were detected by microplate reader.Reactive oxygen species(ROS)production was evaluated by fluorescent probes.Bax,Bcl-2 and Caspase-3 protein and mRNA expressions were assessed by Western blotting and RT-qPCR.Phosphorylation levels of ERK,JNK and p38MAPK proteins were assessed by Western blotting.Results The results indicated that,cells were injured by sitmulating with 100μmol/L H2O2 for 2hrs,TFs and cardamonin have the antioxidant effects against H2O2 induced oxidative injury.Flavonoids prevented cytotoxicity and apoptosis in SVARECs,modulated the enzymes of oxidation,attenuated excessive ROS generation,reduced caspase-3 expression and Bax/Bcl-2 protein ratio,increased the Phosphorylation levels of Akt and decreased the Phosphorylation levels of JNK and p38MAPK.Conclusion TFs extracted from LCC and cardamonin itself showed protective effects against oxidative injury and could be developed as an effective and potential candidate to prevent oxidative stress in the future. |
