Experimental study on EGCG delaying endothelial cell senescence via Nrf2/HO-1 signaling pathway
ZHANG Ying-jie
WANG Ling-huan
MA Yan
WANG Tian-hu
GU Zheng-hui
FANG Zhi-yi
SHEN Xiao-ying
YUAN Yuan
TIAN Lei
CAO Feng
Abstract：Objective To investigate the effects and mechanisms of(-)-epigallocatechin-3-gallate(EGCG)on doxorubicin(Dox)-induced senescence in human umbilical vein endothelial cells(HUVEC).Methods Between April and December 2023,HUVEC were cultured in vitro at the Geriatric Research Institute of the Second Medical Center,PLA General Hospital.The cells were divided into four groups:the control group,the Dox group(2 μmol/L doxorubicin),the Dox+EGCG group(50 μmol/L epigallocatechin gallate)and the Dox+EGCG+ML385 group(with 2.5 μmol/L ML385,a specific Nrf2 inhibitor).Cellular senescence was assessed usingβ-galactosidase(β-gal)staining;intracellular reactive oxygen species(ROS)levels were measured using ROS staining;kits for superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were used to assess SOD activity and MDA levels;cell viability was evaluated using CCK-8 assay;levels of senescence-related proteins(P21 and P53),nuclear factor-E2-related factor 2(Nrf2)and heme oxygenase 1(HO-1)were detected by Western blot.Results Treatment with EGCG at concentrations of 10,30,and 50 μmol/L for 48 h did not affect the cell viability,while cell viability decreased after pretreatment with EGCG at concentrations of 70,100,and 300μmol/L,thus 50 μmol/L was selected as the subsequent concentration.Compared with the control group,the Dox group showed a significant increase in β-gal positivity and ROS levels(P＜0.05),elevated expression of P21 and P53 proteins(P＜0.05).After the addition of EGCG,β-gal positivity and ROS levels were significantly reduced(P＜0.05),but SOD activity increased(P＜0.05)and MDA content decreased(P＜0.05).Additionally,the expression levels of P21 and P53 proteins were reduced(P＜0.05),while Nrf2 and HO-1 levels were significantly increased P＜0.05).The ML385 group reversed the anti-senescence effect of EGCG,manifested by decreased expression of Nrf2 and HO-1(P＜0.05)and increased expression of P21 and P53 proteins(P＜0.05).Conclusion EGCG may delay dox-induced senescence in HUVEC by activating the Nrf2/HO-1 pathway.
Keywords：(-)-epigallocatechin-3-gallateHuman umbilical vein endothelial cellsSenescenceNrf2HO-1
Publication Date：2024-06-10
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 571-576 )