The Effect Of Resveratro1 On HO-1 Expression In Human Mesangial Cells Cultured With High Glucose

Objective To investigate the effect of resveratrol on heme oxygenase-1 (HO-1) expression and proliferation of human mesangial cells (HMCs) cultured with high glucose.Methods HMCs were cultured under DMEM with normal glucose to 80% confluence, washed once with serum-free DMEM, and growth-arrest in serum-free DMEM in NG for 24 hours to synchronize the cell growth.After this time, HMCs were divided into normal glucose(NG group, 5.6mmol/L),middle glucose(MG group,11.2mmol/L), high glucose (HG group,22.4mmol/L), high glucose plus resveratrol at 5,10,20,40,80μmol/L respectively definition is HG+Resl group, HG+Res2 group, HG+Res3 group, HG+Res4 group, HG+Res5 group and high glucose plus resveratrol and zinc protoporphyrin(HG+Res+Znpp group, resveratrol 20μmol/L,Znpp 10μmol/L), high glucose plus resveratrol and Rapamycin(HG+ Res+Rapa group, resveratrol 20μmol/L, Rapa 10μmol/L).Collecting HMCs of NG, MG, HG group at 6,12,24,48h, Collecting HMCs of HG+Resl, HG+Res2, HG+Res3, HG+Res4, HG+Res5, HG+ Res+Znpp and HG+ Res+Rapa group at 48h,the expression of HO-1 and mTOR mRNA were observed using Real-time PCR. Collecting HMCs of NG, HG, HG+Res3, HG+ Res+Znpp and HG+ Res+Rapa group at 48h, The expression of HO-1 protein were detected using immunofluorescence with laser scanning confocal microscopy.HMCs proliferation ability of NG, HG, HG+Res3, HG+ Res+Znpp group was detected by MTT colorimetry, TGF-β1 levels were measured with ELISA,and TAC,GSH-Px,MDA in supernatants were measured too.Results 1. HO-1mRNA expression:①No changes can be found in NG group at 6h,12h,24h,48h. It increased along with time go by in MG group,and it increased significantly at 24,48h(P<0.05,P<0.01). Compared with NG Group, the expression of HO-1mRNA in HG group was increased at 6,12,24h,but decreased at 48h(P<0.01).②The expression of HO-1mRNA in HG+Res1, HG+Res2 and HG+Res3 group was enhanced compared to HG group,especialy HG+Res2 group (P<0.01),but decreased in HG+Res4, HG+Res5 group(P<0.01).2. The expression of HO-1mRNA and protein in HG group was lower than NG Group(P<0.05,P<0.01),and the mTOR mRNA expression was higher than NG Group(P<0.01). Res increased HO-1 expression but decreased mTOR expression(P<0.01). Compared with HG+Res group, HO-1 expression was not changed in HG+Res+Rap group,but mTOR expression was decreased(P<0.01).3. The proliferation activity of HMCs,the lever of TGF-(31 and MDA in HG group all were higher than that in NG group(P<0.01),but GSH-Px activity and TAC lever was decreased in HG group(P<0.01). After treatmenting HMCs with resveratrol, HMCs proliferation, TGF-01 and MDA lever was decreased and GSH-Px activity,TAC was increased compared to HG group (P<0.01). Compared with HG+Res group, HG+Res+Znpp group showed a significant increase of HMCs proliferation, TGF-β1 and MDA lever(P<0.01),but was still lower than HG group,with an decrease of GSH-Px activity and TAC lever (P<0.01), but TAC lever was still higher than HG group.Conclusion 1. The expression of HO-1 compensatory increased at early time,then suppressed in HMCs cultured with high glucose. Moderate resveratrol could upregulate HO-1 expression in HMCs cultured with high glucose.2. Resveratrol could inhibit HMCs proliferation, TGF-β1 and oxidative stress lever caused by high glucose through inducing HO-1 expression.