APPL1 ameliorates myocardial ischemia‑reperfusion injury by regulating the AMPK signaling pathway

Abstract

Myocardial ischemia-reperfusion injury results in elevated reactive oxygen species (ROS) production and causes oxidative stress damage. Therefore, the current study aimed to investigate whether adaptor protein phosphotyrosine interacting with PH domain and leucine zipper 1 (APPL1) could induce the expression of antioxidant enzymes through AMP-activated protein kinase (AMPK) signaling in order to alleviate the injury caused by ischemia/hypoxia-reperfusion. Following induction of hypoxia-reoxygenation (H/R) injury in H9c2 cells, the liver kinase B1 (LKB1)/AMPK/acetyl-CoA carboxylase α (ACC) signaling pathway was investigated using western blot analysis, along with the detection of superoxide dismutase (SOD)2 and SOD3 expression. Additionally, cell viability was detected using a Cell Counting Kit-8 assay and ROS production was analyzed using ROS staining, whereas the expression levels of inflammatory mediators (TNF-α, monocyte chemoattractant protein 1 and IL-1β), apoptosis mediators [cleaved caspase-3, cleaved poly (ADP-ribose) polymerase and Bcl-2] and nuclear factor erythroid 2-related factor 2 signaling pathway-related proteins were detected via western blot analysis following overexpression of APPL1 alone or in combination with compound C treatment (an AMPK inhibitor). The results indicated that H/R induction upregulated the phosphorylation levels of LKB1, AMPK and ACC, and decreased the expression levels of APPL1 and SOD enzyme activities. APPL1 overexpression increased the phosphorylation levels of LKB1, AMPK and ACC, SOD enzyme activity and cell viability whereas the expression levels of proinflammatory mediators and proapoptotic mediators, and the levels of ROS production were markedly decreased when compared with H/R group with empty plasmid transfection. APPL overexpression-mediated effects were significantly abrogated by compound C. Taken together, the data indicated that APPL1 inhibited ROS production and H/R-induced myocardial injury via the AMPK signaling pathway. Therefore, APPL1 may serve as a potential therapeutic target for myocardial H/R injury.

METTL14-mediated N6-methyladenosine modification induces the ferroptosis of hypoxia/reoxygenation-induced cardiomyocytes

“…4 A). Among intersections, the downregulation of APPL1 after H/R treatment has been reported previously [ 31 , 32 ]. Therefore, the dual-luciferase assay was conducted according to the binding site (Fig.…”

“…APPL1 is known to protect against cardiomyocyte senescence, cardiac fibrosis, and diabetic cardiomyopathy [ 61 – 63 ]. More importantly, APPL1 can function as a potential therapeutic target for myocardial I/R injury by antagonizing apoptosis, oxidative stress, cytotoxicity, and release of inflammatory cytokines [ 31 , 32 , 64 ]. In the same light, our experimentation uncovered that the downregulation of APPL1 mRNA was found in H/R-induced cardiomyocytes and miR-146a-5p mimics potently repressed APPL1 transcription.…”

“…Adaptor protein phosphotyrosine interacting with PH domain and leucine zipper 1 (APPL1) is a vital modulator of insulin and adiponectin signaling [ 30 ]. By interacting with AMP-activated protein kinase signaling or binding to adiponectin receptor, APPL1 can attenuate H/R-induced apoptosis, inflammation, and ROS generation in myocardial I/R injury [ 31 , 32 ]. miRNAs can induce mRNA decay through recognition of the 3’ untranslated region and APPL1 was identified as a target gene of miR-146a-5p using multiple databases, suggesting the regulatory role of miR-146a-5p in APPL1 biogenesis in myocardial I/R injury.…”

Serelaxin Protects H9c2 Cardiac Myoblasts against Hypoxia and Reoxygenation-Induced Damage through Activation of AMP Kinase/Sirtuin1: Further Insight into the Molecular Mechanisms of the Cardioprotection of This Hormone

“…To investigate the involvement of AMPK signaling in the possible stimulation of SIRT1 by RLX, H9c2 cells were treated for 24 h before the onset of hypoxia with 10 and 20 µmol L −1 of Compound C (Dorsomorphin) (MedChemExpress, Monmouth Junction, NJ, USA), a selective and ATP-competitive AMPK inhibitor. The chosen concentrations and exposure time of Compound C were as reported in [29].…”