Prelimilary Investigation For The Effects Of -SH Injury On Dysfunction Of NO Pathway In Endothelial Cells And Its Possible Mechanism

Objective: To investigate the possible mechanism of oxidative stress-induced NO pathway dysfunction, which may involve in the pathogenesis of atherosclerosis,by treating endothelial cells with diethyl maleate(DEM) for interfering intracellular bioactivity of dimethylarginine dimethy laminohy-drolase(DDAH). Methods: Endothelial cells ECV304 were first treated with different concentration of DEM for different time to select a optimal DEM concentration and action time which can obviously decline the intracellular GSH content without significant impact on cell's viability. The cell's viability was detected by MTT assay . Endothelial cells were then Pretreated with the selected dosage of DEM, after pretreatment and washing for removing DEM, the cells incubated further for 0,0.5,2,6,12,24 hours with serum-free medium. The intracellular GSH content,DDAH activity and NO content were measured at the six time points, control groups were set for every measurement. Results: The MTT results showed that the optimal DEM treating concentration is 0.6μmol/L, treating time is 0.5 h. Compared with control group, the intracellular GSH content decreased by 28.7%(p<0.05 ) at 0.5 h after DEM treatment, then it gradually rebounded , and even turned higher than the basal level at 12 h, at 24 h the GSH content returned to basal...