Downregulation of microRNA‑199a‑5p attenuates hypoxia/reoxygenation‑induced cytotoxicity in cardiomyocytes by targeting the HIF‑1α‑GSK3β‑mPTP axis

Abstract

MicroRNAs (miRs) have been identified as critical regulatory molecules in myocardial ischemia/reperfusion injury; however, the exact expression profile of miR-199a-5p in reperfusion injury and the underlying pathogenic mechanisms remain unclear. In the present study, it was revealed that miR-199a-5p expression was significantly increased in the plasma of patients with acute myocardial infarction and in a H9c2 cell model of oxygen-glucose deprivation and reperfusion (OGD/R) via reverse transcription-quantitative PCR. H9c2 cells were transfected with miR-199a-5p mimic or inhibitor, or short interfering RNA (siRNA) specific to hypoxia-inducible factor-1α (HIF-1α). MTS, lactate dehydrogenase (LDH), TUNEL staining and flow cytometry assays were performed to determine the proliferation, LDH activity, apoptosis and mitochondrial membrane potential (ΔΨm) of H9c2 cells, respectively. The overexpression of miR-199a-5p in the OGD/R cell model significantly decreased the viability and increased the lactate dehydrogenase leakage of cells; whereas knockdown of miR-199-5p induced the opposing effects. Additionally, inhibition of miR-199-5p significantly attenuated OGD/R-induced alterations to the mitochondrial transmembrane potential (ΔΨm) and increases in the apoptosis of cells. Furthermore, the overexpression or knockdown of miR-199a-5p decreased or increased the expression of HIF-1α and phosphorylation of glycogen synthase kinase 3β (GSK3β) in OGD/R-treated H9c2 cells. Additionally, siRNA-mediated downregulation of HIF-1α decreased phosphorylated (p)-GSK3β (Ser9) levels and reversed the protective effects of miR-199a-5p inhibition on OGD/R-injured H9c2 cells. Similarly, treatment with LiCl (a specific inhibitor of p-GSK3β) also attenuated the protective effects of miR-199a-5p knockdown on OGD/R-injured H9c2 cells. Mechanistic studies revealed that HIF-1α was a target of miR-199a-5p, and that HIF-1α downregulation suppressed the expression of p-GSK3β in OGD/R-injured H9c2 cells. Furthermore, an miR-199a-5p inhibitor increased the interaction between p-GSK3β and adenine nucleotide transferase (ANT), which was decreased by OGD/R. Additionally, miR-199a-5p inhibitor reduced the OGD/R-induced interaction between ANT and cyclophilin D (Cyp-D), potentially leading to the increased mitochondrial membrane potential in inhibitor-transfected OGD/R-injured H9c2 cells. Collectively, the present study identified a novel regulatory pathway in which the upregulation of miR-199a-5p reduced the expression of HIF-1α and p-GSK3β, and potentially suppresses the interaction between p-GSK3β and ANT, thus promoting the interaction between ANT and Cyp-D and potentially inducing cytotoxicity in OGD/R-injured H9c2 cells.

The miRNA199a/SIRT1/P300/Yy1/sST2 signaling axis regulates adverse cardiac remodeling following MI

“…Cardiac adverse hypertrophy remains the major risk factor for heart failure, which represents one of the leading causes of re-hospitalization and death in general population 17 , 19 . In this context, cardiomyocyte necrosis and mechanical stretching cause the release of circulating soluble sST2 isoform 8 , 20 , 21 , a key biomarker used to diagnose cardiac disease conditions accurately and to effectively prognosticate the high-risk patients.…”

“…Indeed, this up-regulation increases apoptosis 15 and inhibits cell proliferation 16 . Pharmacological inhibition of miR-199-5p attenuates hypoxia/reoxygenation-induced cytotoxicity and decreases phenylephrine-induced cardiomyocyte hypertrophy 17 , 18 , indicating that the inhibition of this miRNA might serve as a novel therapeutic approach for treating HF. Studies specifically designed to assess the potential role and mechanism of miR-199a in the treatment of HF are necessary.…”

Evidence for the Capability of Roxadustat (FG-4592), an Oral HIF Prolyl-Hydroxylase Inhibitor, to Perturb Membrane Ionic Currents: An Unidentified yet Important Action

“…Because of the aforementioned considerations, rising attempts were made to evaluate whether roxadustat or other related compounds could produce any modifications on various types of ionic currents (e.g., delayed-rectifier K + current [ I K (DR ) ], erg -mediated K + current [ I K(erg) ], and voltage-gated Na + current [ I Na ]) in pituitary tumor (GH 3 ) cells. Moreover, earlier numerous studies have reported the effective regulations of HIF-1α expression present in heart-derived H9c2 myoblasts [24,28,29,30,31,32,33,34,35]. Thus, we further decided to investigate the possible actions of roxadustat on I K(DR) in these cells.…”

Downregulation of miRNA‑663b protects against hypoxia‑induced injury in cardiomyocytes by targeting BCL2L1

“…Zhang et al (2) also revealed that miR-103a-3p serves a protective role in myocardial I/R injury. Liu et al (28) demonstrated that miR-199a-5p expression affects hypoxia/reoxygenation-induced cytotoxicity in cardiomyocytes. Additionally, Peng et al (20) demonstrated that miR-133, miR-1291 and miR-663b expression was significantly increased in AMI compared with controls.…”