The Protective Effect Of Resveratrol On High Glucose Induced Oxidative Damage In Human Umbilical Ein Endothelial Cells

Objective To investigate the protective effect of Res to oxidative stress and proliferation of human umbilical vein endothelial cell in high glucose circumstance.Methods Human umbilical vein endothelial cells(ECV304)were cultured in vitro.The cultured cells were then divided to normal glusoce(NG group 5.6mmol/L), high glucose(HG group,30mmol/L),various concentrations of Res(NG+Res10^-3-mol/L-10^-5mol/L),and HG group was treated with Res(HG+Res10^-3mol/L-10^-5mol/L )when were cultured at 24 hours.Cell proliferation ability was tested,concertration of NO,ET-1 and ICAM-1 in supernatant were measured in NG group,HG group at 24, 48 and 72hours;Cell proliferation ability was tested in NG+Res10^-3mol/L-10^-5mol/L, and HG+Res10^-3mol/L-10^-5mol/L when were cultured at 48 hours,concertration of NO,ET-1 and ICAM-1 in supernatant were measured in high glucose was treated with Res10^-4mol/L.The level of SOD,TAC,NOS,iNOS and MDA in cell were also determined in NG group,HG group,HG+Res10^-4mol/L.The expression of NF-κB₆₅ and iNOS mRNA were observed using RT-PCR.Meanwhile,Western-blot method was employed to half-quantitatively determine the NF-κB₆₅expression.Results 1.The cell proliferation of HG group was significantly inhibited at 48 and 72hours compared with NG group(P＜0.01,P＜0.05),but no changes can be found between the groups of various concentrations of Res and NG group; HG+Res10^-4mol/L,10^-5mol/L had increased the value of MTT as compared HG group(P＜0.01),but still had lower than NG group;HG+Res10^-4mol/L higher than HG+Res10^-4mol/L.2.Meanwhile,HG group had higher NO,ET-1,ICAM-1 level than NG group(P＜0.05,P＜0.01,P＜0.01),HG+Res had lower than HG group.The lever of ET-1 was near NG group,but ICAM-1 still higher than NG group(P＜0.01).3. Compared with NG group,HG group showed a significant decrease of SOD activity and TAC level(P＜0.05),with an increase of MDA(P＜0.01),When HG + Res,SOD activity and TAC improved(P＜0.05),and SOD,TAC lower than NG group,while MDA was near to NG group.4.Compared with NG group,HG group showed a significant decrease of NOS,iNOS,eNOS level(P＜0.01,P＜0.05,P＜0.01),When HG + Res,while NOS,iNOS,eNOS(P＜0.01,P＜0.05,P＜0.01)level went down.NOS and iNOS was near to NG group,iNOS had increased as compares with NG group (P＜0.05).5.When it came to the mechanism,both NF-κB₆₅and iNOS mRNA expression,the half-quantitatively of protein about NF-κB₆₅increased in HG group (P＜0.01).However,Res could decrease the expression of these two mRNAs,then higher than NG group(P＜0.05).The expression alteration of NF-κB₆₅was positively correlated with that of iNOS(r=0.554,P＜0.01).The alteration of NF-κB₆₅was positively correlated with that of ICAM-1(r=0.356,P＜0.01).Conclusions 1.High glucose environment 1ed to oxidative stress of human umbilical vein endothelial cell and thus inhibited its proliferation.Res treatment could significantly resist the oxidative stress and proliferation caused by high glucose,but have no effect to normal human umbilical vein endothelial cell.2.One possible mechanism of Res effect could be inhibition of NF-κB₆₅and iNOS mRNA expression so that ICAM-1 and ET-1 decreased,NO was adjusted.