Protective Effects Of L-arginine And Curcumine On ECV-304 Incubated With Diabetic Serum

Objective To investigate the injury induced by diabetic serum(DS) to the cultured humen umbilical vein endothelial cells(ECV-304) and the protective effects of L-arginine(L-Arg) and curcumine(CC) on them and to search for the possible mechanism.Methods The cultured ECV-304 cells were divided into 4 groups and 10% serum of patient with only type 2 diabets mellitus(T2DM)(DS1), 10% serum of patient with diabetic macrovascular complications(DS2), 10% serum of healthy subject(HS) and 10% normal sodium(NS) were added to the culture medium. After 30 minutes, 3 hours and 3 days, the levels of nitric oxide(NO), malondialdehyde(MDA) and lactate dehydrogenase(LDH) in the culture medium were determined and cells proliferative activity was measured by MTT assay after 3 days.Then atovastatin(AT)( 1, 5, 10μM), L-Arg (0.5, 1.0, 1.5mM ), CC(5, 10, 15μM) and 0.1% dimethyl sulfoxide(DMSO) were respectively added to ECV-304 cells treated with the culture medium including 10% DS1. After 3 days, cells proliferative activity was measured by MTT assay. Both groups of DS1 and DS2 had no difference in injury to ECV-304, so treatment with DS1 acted as DS control group.Lastly, after endothelial injury was induced by 10% DS, AT, L-Arg, CC (in the most effective concentration) and 0.1%DMSO were respectively added to the medium. After 30 minutes, 3 hours and 3 days, cells viability was measured by trypan blue exclusion method and the levels of NO, MDA and LDH in the medium were detected.Results The results of MTT showed that DS1(P<0.05) and DS2(P<0.01) decreased the cells proliferative activity in comparison with NS control group.