MicroRNA-668-3p Protects Against Oxygen-Glucose Deprivation in a Rat H9c2 Cardiomyocyte Model of Ischemia-Reperfusion Injury by Targeting the Stromal Cell-Derived Factor-1 (SDF-1)/CXCR4 Signaling Pathway

Abstract

Departmental sources Background: Ischemia-reperfusion injury (IRI) results from the restoration of blood supply to ischemic organs, including the heart. Expression of microRNA-668-3p (miR-668-3p) is known to protect the kidney from IRI. This study aimed to investigate the role of miR-668-3p in oxygen-glucose deprivation (OGD) in a rat H9c2 cardiomyocyte model of IRI. Material/Methods: Rat H9c2 cardiomyocytes were cultured in glucose-free Dulbecco's modified Eagle's medium (DMEM) under anaerobic conditions, followed by oxygenation, to create the OGD model of IRI. The luciferase reporter assay evaluated the interaction between stromal cell-derived factor-1 (SDF-1), or CXC motif chemokine 12 (CXCL12), and miR-668-3p. Protein and mRNA levels of SDF-1, CXCR4, Bcl2, Bax, cleaved caspase-3, endothelial nitric oxide synthase (eNOS), and phosphorylated endothelial nitric oxide synthase (p-eNOS) were analyzed by Western blot and quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and apoptosis were assessed by flow cytometry. Enzyme-linked immunosorbent assay (ELISA) measured reactive oxygen species (ROS), including malondialdehyde (MDA), nitric oxide (NO), p-eNOS, and the inflammatory cytokines, tumor necrosis factor-a (TNF-a), interleukin-1b (IL-1b), IL-6, and monocyte chemoattractant protein-1 (MCP-1) in H9c2 cell supernatants. Results: In the OGD rat H9c2 cardiomyocyte model of IRI, miR-668-3p levels were reduced. Overexpression of miR-668-3p inhibited SDF-1, CXCR4, the expression of inflammatory cytokines, markers of oxidative stress, and p-eNOS. The overexpression of SDF-1 reversed these findings. Overexpression of SDF-1 promoted cell apoptosis, which was reduced by miR-668-3p. Conclusions: In the OGD rat H9c2 cardiomyocyte model of IRI, miR-668-3p suppressed mediators of inflammation and oxidative stress and enhanced cell viability through the SDF-1/CXCR4 signaling pathway.

Cardiac protection induced by urocortin-2 enables the regulation of apoptosis and fibrosis after ischemia and reperfusion involving miR-29a modulation

“…The selection of these genes was based on their fold change rates as well as their implication in I/R-related processes, as published elsewhere. 20 , 21 , 22 Namely, we investigated the expression of Apoptosis Inducing Factor mitochondria associated 1 ( Aifm1 ), Apoptosis Protease-Activating Factor-1 ( Apaf1 ), B-Cell Lymphoma 2 ( Bcl-2 ), Cytochrome c ( Cycs ), Heme Oxygenase 1 ( Hmox-1 ), and Mitogen-Activated Protein Kinase 8 ( Mapk-8 ). Figures 6 A and 6B show that the expression of Apaf-1 and Hmox-1 increased significantly in I/R, as compared with controls.…”

microRNA-9-5p protects liver sinusoidal endothelial cell against oxygen glucose deprivation/reperfusion injury

“…However, whether such binding exerts a functional role needs to be tested. CXCR4 is a crucial mediator of hepatic, cardiac, and lung ischemia-reperfusion injury [ 40 , 41 , 42 ]. In the present study, we observed that the reduction of miR-9-5p correlated with enhanced expression of CXCR4 mRNA and protein levels.…”

The Role of Hypoxia-Inducible Factor-1 Alpha in Renal Disease

“…In addition, the intrinsic mechanisms implicated in RIRI-associated cell death include the disruption of mitochondrial dynamics, resulting in mitochondrial fragmentation. Wei et al revealed that microRNA-668 (miR-668), previously found up-regulated in the RIRI model, has a protective effect during RIRI by inhibiting pathogenic mitochondrial fragmentation and the subsequent apoptosis of renal tubular cells [ 28 ]. Moreover, the induction of miR-668 in RIRI is HIF-1 dependent, while among the miR-688 target genes, only mitochondrial protein 18kDa (MTP18) is associated with mitochondrial dynamics.…”