Inhibition of microRNA‑124‑3p protects against acute myocardial infarction by suppressing the apoptosis of cardiomyocytes

Hu,Guangrong; Ma,Lingbo; Dong,Fei; Hu,Xiao; Liu,Sida; Sun,Hui

doi:10.3892/mmr.2019.10565

Inhibition of microRNA‑124‑3p protects against acute myocardial infarction by suppressing the apoptosis of cardiomyocytes

Authors:
- Guangrong Hu
- Lingbo Ma
- Fei Dong
- Xiao Hu
- Sida Liu
- Hui Sun
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Affiliations: Emergency Department, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China
Published online on: August 6, 2019 https://doi.org/10.3892/mmr.2019.10565
Pages: 3379-3387

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Abstract

The aims of the present study were to investigate the roles and underlying mechanisms of microRNA‑124‑3p (miR‑124‑3p) in the progression of acute myocardial infarction (AMI). The expression of miR‑124‑3p was determined via reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). TargetScan analysis and a luciferase reporter assay were conducted to reveal the association between miR‑124‑3p and nuclear factor κ‑light‑chain‑enhancer of activated B cells (NF‑κB) repressing factor (NKRF). To investigate the role of miR‑124‑3p in AMI, a cell model of myocardial hypoxic/ischemic injury was established by subjecting H9c2 cardiac cells to hypoxia for 48 h. The viability of cells was determined using an MTT assay, and cell apoptosis was analyzed by flow cytometry. Additionally, the expression levels of inflammatory factors [tumor necrosis factor‑α (TNF‑α), interleukin (IL)‑1β and IL‑6] were measured via ELISA. Furthermore, gene and protein expression levels were determined by performing RT‑qPCR and western blot analyses, respectively. It was revealed that the expression of miR‑124‑3p was significantly increased in the blood of patients with AMI and hypoxia‑treated H9c2 cells. Additionally, it was demonstrated that NKRF was a direct target of miR‑124‑3p. The hypoxia‑induced decrease in the viability of H9c2 cells and increase in cell apoptosis were eliminated by the downregulation of miR‑124‑3p. Furthermore, hypoxia significantly increased the levels of TNF‑α, IL‑1β and IL‑6, whereas miR‑124‑3p downregulation eliminated these effects. Downregulated expression of B‑cell lymphoma 2, pro‑caspase 3 and pro‑caspase 9 protein, and upregulated expression of cleaved caspases 3 and 9 was observed in hypoxic H9c2 cells; the altered expression of these proteins was suppressed by miR‑124‑3p inhibitor. Additionally, miR‑124‑3p inhibitor suppressed the hypoxia‑induced activation of the NF‑κB signaling pathway in H9c2 cells. Furthermore, it was demonstrated that the various effects of miR‑124‑3p inhibitor on H9c2 cells were eliminated by the small interfering RNA‑mediated downregulation of NKRF. In conclusion, the results of the present study indicated that miR‑124‑3p downregulation protected against AMI via inhibition of inflammatory responses and the apoptosis of cardiomyocytes by regulating the NKRF/NF‑κB pathway.

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