心力衰竭(HF)是一种慢性疾病,其中心脏无法向外周组织提供足够的血液和氧气。心肌细胞凋亡和自噬与HF进展有关。然而,HF的潜在机制是未知的。在这项研究中,H2O2处理的AC16细胞用作HF的细胞模型。使用RT-qPCR和Westernblot检测相关基因的mRNA和蛋白水平。使用CCK-8和流式细胞术评估细胞活力和凋亡,分别。ETS2,TUG1,miR-129-5p,和ATG7通过荧光素酶活性验证,ChIP,和RNA结合蛋白免疫沉淀测定。根据我们的发现,H2O2刺激增加了AC16细胞中ETS2,TUG1和ATG7的表达,同时降低了miR-129-5p的表达。此外,H2O2刺激诱导心肌细胞凋亡和自噬,被ETS2耗尽逆转,TUG1消音,或miR-129-5p上调。机械上,ETS2通过与TUG1启动子结合促进TUG1表达,和TUG1海绵miR-129-5p增加ATG7表达。此外,TUG1过表达逆转了ETS2敲低介导的心肌细胞凋亡和自噬抑制,miR-129-5p抑制消除了TUG1耗竭介导的H2O2诱导的AC16细胞对心肌细胞凋亡和自噬的抑制。据推测,在H2O2诱导的AC16细胞中,ATG7过表达逆转miR-129-5p模拟介导的心肌细胞凋亡和自噬的抑制。最后,ETS2沉默通过靶向ETS2/TUG1/miR-129-5p/ATG7轴来减少心肌细胞凋亡和自噬以减缓HF进展,这可能为HF治疗提供新的治疗靶点。
Heart failure (HF) is a chronic disease in which the heart is unable to provide enough blood and oxygen to the peripheral tissues. Cardiomyocyte apoptosis and autophagy have been linked to HF progression. However, the underlying mechanism of HF is unknown. In this study, H2 O2 -treated AC16 cells were used as a cell model of HF. The mRNA and protein levels of related genes were examined using RT-qPCR and western blot. Cell viability and apoptosis were assessed using CCK-8 and flow cytometry, respectively. The interactions between ETS2, TUG1, miR-129-5p, and ATG7 were validated by luciferase activity, ChIP, and RNA-Binding protein Immunoprecipitation assays. According to our findings, H2 O2 stimulation increased the expression of ETS2, TUG1, and ATG7 while decreasing the expression of miR-129-5p in AC16 cells. Furthermore, H2 O2 stimulation induced cardiomyocyte apoptosis and autophagy, which were reversed by ETS2 depletion, TUG1 silencing, or miR-129-5p upregulation. Mechanistically, ETS2 promoted TUG1 expression by binding to the TUG1 promoter, and TUG1 sponged miR-129-5p to increase ATG7 expression. Furthermore, TUG1 overexpression reversed ETS2 knockdown-mediated inhibition of cardiomyocyte apoptosis and autophagy and miR-129-5p inhibition abolished TUG1 depletion-mediated suppression of cardiomyocyte apoptosis and autophagy in H2 O2 -induced AC16 cells. As presumed, ATG7 overexpression reversed miR-129-5p mimics-mediated repression of cardiomyocyte apoptosis and autophagy in H2 O2 -induced AC16 cells. Finally, ETS2 silencing reduced cardiomyocyte apoptosis and autophagy to slow HF progression by targeting the ETS2/TUG1/miR-129-5p/ATG7 axis, which may provide new therapeutic targets for HF treatment.