Molecular mechanism of neural epithelial transforming factor-1 improving apoptosis in myocardial ischemia-reperfusion injury through regulating nuclear factor-κB/β-catenin signaling pathway
LI Jiaxin
ZHENG Wei
SONG Mingxing
SONG Xiurong
LI Hongyu
LI Meng
Abstract：Objective To elucidate the molecular mechanism of neural epithelial transforming factor-1(NET-1)improving apoptosis in myocardial ischemia-reperfusion injury(MIRI)through regulating nuclear factor-κB(NF-κB)/β-catenin signaling pathway.Methods An in vitro model of ischemia-reperfusion(I/R)injury in H9C2 cells was established via hypoxia/reoxygenation(H/R)treatment.The experimental design consisted of two parts:(1)A hypoxia time-gradient experiment(0,2,4,6,8,12 h)combined with gene overexpression/knockdown techniques to investigate the correlation between cell apoptosis and NET-1 expression.(2)Mechanistic studies:Co-immunoprecipitation(Co-IP)was performed to verify the protein-protein interactions between NET-1 and NF-κB,as well as between NF-κB and β-catenin.Furthermore,Western blot and RT-qPCR assays were employed to analyze the expression of downstream target genes,in conjunction with gene overexpression/knockdown techniques.Results(1)Cardiomyocyte apoptosis was positively correlated with hypoxia duration and NET-1 expression,with the peak observed at 6 h of hypoxia exposure.NET-1 overexpression significantly exacerbated cell apoptosis,whereas NET-1 knockdown markedly suppressed apoptosis.With the extension of hypoxia duration,the protein and mRNA expressions of NET-1,Caspase-3 and Bax were significantly upregulated,peaking at 6 h.Moreover,the protein and mRNA expressions of Caspase-3 and Bax were significantly increased in NET-1 overexpression group,but prominently decreased in NET-1 knockdown group.(2)NF-κB exerted a pro-apoptotic effect in hypoxia/reoxygenation-induced cardiomyocyte apoptosis,and it could interact with and be regulated by NET-1.The expression level of NF-κB was positively correlated with the protein and mRNA expressions of Caspase-3 and Bax.The expression of NF-κB was significantly increased in NET-1 overexpression group,but markedly decreased in NET-1 knockdown group.Co-IP assays demonstrated that NET-1 could be co-precipitated with NF-κB.(3)β-catenin exerted a pro-apoptotic effect in hypoxia/reoxygenation-induced cardiomyocyte apoptosis,and it could interact with and be regulated by NF-κB.The expression level of β-catenin was positively correlated with the protein and mRNA expressions of Caspase-3 and Bax.The expression of β-catenin was significantly increased in NF-κB overexpression group,but markedly decreased in NF-κB knockdown group.Co-IP assays demonstrated that NF-κB could be co-precipitated with β-catenin.Conclusion NET-1 positively regulates NF-κB and activates β-catenin,thereby synergistically promoting cardiomyocyte apoptosis associated with MIRI.Targeted inhibition of NET-1 can significantly alleviate cardiomyocyte apoptosis,suggesting its potential value as a novel therapeutic target.
Keywords：neural epithelial transforming factor-1myocardial ischemia-reperfusion injurymyocardial apoptosisnuclear factor-κBβ-catenin
Publication Date：2026-02-20
Online Publishing Date：2026-03-19(First online date of this platform, not the publication date of the document)
Pages：7( 210-216 )