Resveratrol Attenuates Senescence Of Human Umbilical Vein Endothelial Cells （HUVECs） Induced By H₂O₂ Combined With Trimethylamine-N-oxide And Its Potential Mechanisms

Objectives To detect the effects of resveratrol（RSV）,H₂O₂ and trimethylamine-N-oxide（TMAO）on the senescence of human umbilical vein endothelial cells（HUVECs）,and to explore whether RSV can attenuate the senescence of HUEVCs and its potential mechanisms.Methods HUVECs were cultured and divided into six groups:control,H₂O₂,TMAO,RSV,H₂O₂+TMAO and RSV+H₂O₂+TMAO according to the intervention for 72h,and RSV was pretreated for 12h.SA-β-Gal staining was used to detect the senescent cells and Western Blot was applied to detect the expressions of senescence-associated proteins p53,PAI-1 and p21 and related pathway proteins AMPK,p-AMPK,SIRT1,FOXO1 and eNOS.Results（1）The percentage of SA-β-Gal positive cells and the expressions of p53,PAI-1 and p21 on HUVECs in the H₂O₂ group were higher than those in the control,TMAO and RSV group（p<0.05 for all）,but there was no significant change in the TMAO and RSV group（p<0.05 for all）,while the above indicators in the H₂O₂+TMAO group were significantly higher than those in the control,H₂O₂,TMAO and RSV group respectively（p<0.05 for all）.（2）The percentage of SA-β-Gal positive cells and the expressions of p53,PAI-1 and p21 on HUVECs in the RSV+H2O2+TMAO group were lower than those the in H₂O₂+TMAO group（p<0.05 for all）.（3）The expressions of p-AMPK,SIRT1,FOXO1 and eNOS on HUVECs in the H₂O₂,TMAO and H₂O₂+TMAO group were lower than those in the control group,and the H₂O₂+TMAO group was the most obvious（p<0.05 for all）.The expressions of these proteins on HUVECs in the RSV group was higher than those in the control group,and the above changes in the RSV+H2O2+TMAO group were significantly higher than those in the H₂O₂+TMAO group（p<0.05 for all）.There was no significant difference in AMPK expression among the groups（p>0.05）.Conclusions TMAO treatment with HUVECs for 72h does not cause cell senescence,however,it can increase HUVECs senescence induced by H₂O₂.RSV can attenuate HUEVCs senescence promoted by H₂O₂ combined with TMAO intervention,and its possible mechanisms may be associated with activation of AMPK/SIRT1/FOXO1signal pathway and up-regulation of eNOS.