Isorhamnetin Protective Effect On Hydrogen Peroxide Induced Endothelial Cell Injury

Objective:The study was to observe antiapoptotic effect and its mechanism of isorhamnetin acid on human umbilical vein endothelial cell apoptosis induced by H2O2。Method:culture the endothelial cell in vitro,Use the MTT method to sieve the best H2O2 concentration,On the basis of the model , Determination by MTT isorhamnetin endothelial cells relative to the impact of vitality; endothelial cells detected by ELISA medium t-PA and PAI-1 levels;RT-PCR of PAI-1 mRNA expression; Endothelial cells detected by ELISA and medium vWf and TM content; Flow cytometry of endothelial cell necrosis rate and the average fluorescence intensity of the change;Spectrophotometric Determination of fluorescence endothelial cells in the culture medium LDH content; Western Blot method of endothelial cells caspase-3 activity; Confocal laser scanning microscopy of intracellular free calcium concentration changes. Results:compared to the model group:Isorhamnetin can inhibit the secretion of PAI-1, increased t-PA and PAI-1 ratio, And to lower the expression of PAI-1mRNA; isorhamnetin can make medium vWf and TM content, reduce endothelial cell necrosis rate and the average fluorescence intensity LDH in the culture medium to reduce the content; Isorhamnetin can reduce the activity of caspase-3,reduce intracellular free calcium concentration in the rate of increase.Conclusion:isorhamnetin can protect endothelial cells anticoagulant fibrinolytic function, Isorhamnetin can obviously protect endothelial cells, And its protective effect against lipid peroxidation, the restoration and protection of the cell membrane.Inhibition of intracellular calciumion concentration and reduce the increase of caspase-3 activity.