Alleviation of hypoxia/reoxygenation-induced HL-1 cardiomyocyte injury with hydrogen sulfide through regulating Nrf2/HO-1 signaling pathway mediated by PI3K/AKT
Zheng Xiaobin
Wang Chao
Wu Haiyan
Yao Bingqi
Hou Shuxian
Abstract：Objective To investigate the protective effect of hydrogen sulfide(H2S)on hypoxia/reoxygenation(H/R)-inducedHL-1 cardiomyocyte injury,and analyze its potential mechanism.Methods Firstly,selective targeting siRNA of nuclear factor E2 related factor 2(Nrf2)was screened and validated by using RNA interference technology.Then,an H/R injury model of HL-1 cardiomyocytes was established and divided into 5 groups:control group,H/R group,H/R+NaHS group,H/R+NaHS+Nrf2 siRNA group and H/R+Nrf2 siRNA group.The viability of cardiomyocytes was detected by using CCK-8 assay,levels of H2S and malondialdehyde(MDA)and activity of superoxide dismutase(SOD)were detected by using assay kits,and cardiomyocyte apoptosis was observed by using TUNEL staining.Finally,apoptosis,inflammation and protein expressions of Nrf2/heme oxygenase 1(Nrf2/HO-1)signaling pathway and PI3K/AKT signaling pathway were analyzed by using Western blotting assay in all groups.Results The effective siRNA fragments targeting Nrf2 were successfully screened,and their significant inhibitory effects on Nrf2 expression were validated,followed by transfection of Nrf2 siRNA according to grouping requirements.The viability of cardiomyocytes,levels of H2S and SOD,protein expressions of Bcl-2,Nrf2,HO-1,p-PI3K/PI3K,p-AKT/AKT,and ratio of p-GSK-3β/GSK-3β decreased significantly,and MDA,apoptosis severity,protein expressions of cleaved capase-3,Nrf-2,nucleus Fyn and IL-6,and ratio of p-p65/p65 increased significantly in H/R group compared with control group.The pretreatment of NaHS reversed significantly the above changes,but Nrf2 siRNA transfection counteracted partially NaHS protective effect.Conclusion H2S can improve Nrf2 expression and relieve oxidative stress injury induced by H/R inHL-1 cardiomyocytes through activating PI3K/AKT/GSK-3β signaling pathway.
Keywords：Hydrogen sulfideIschemia reperfusion lesionOxidative stressPI3K/AKT signaling pathwayNrf2/HO-1 signaling pathway
Publication Date：2025-01-27
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 32-38 )