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miRNA-146a-5p Inhibits Hypoxia-Induced Myocardial Fibrosis Through EndMT

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Abstract

Cardiac Vascular disease particularly myocardial infarction (MI) is a threat to health worldwide. microRNAs (miRNAs) have been shown to regulate myocardial fibrosis. Therefore, it is potential to investigate the mechanism of miRNA and fibrosis following myocardial infarction. Hypoxia human cardiac microvascular endothelial cells (HCMECs) were selected for the vitro experimental model. The miR-146a-5p expression was tested via RT-qPCR. The level of endothelial-to-mesenchymal transition (EndMT) and fibrosis markers were detected by Western blotting and immunofluorescence. Then, the inflammation, cell viability and apoptosis were investigated. The target was predicted by an online database and verified by a dual-luciferase activity assay. An MI mouse model was created to validate that miR-146a-5p regulates cardiac fibrosis in vivo. MI mouse was transfected with miR-146a-5p lentivirus. Subsequently, its effect on cardiac fibrosis of infarcted hearts was assessed by In situ hybridization (ISH), Immunohistochemistry (IHC), Triphenylterazolium chloride (TTC) staining and Masson staining. Herein, we confirmed that miR-146a-5p was down-regulated in hypoxia HCMECs. Overexpression of miR-146a-5p inhibited hypoxia-induced cardiac fibrosis following myocardial infarction by inhibiting EndMT in HCMECs. Thioredoxin-interacting protein (TXNIP) was a target that was negatively regulated by miR-146a-5p. Up-regulation of miR-146a-5p inhibited cardiac fibrosis via regulating EndMT by targeting TXNIP, and it also regulated EndMT to inhibit cardiac fibrosis in vivo.

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Data Availability

Data sets generated and/or analyzed during the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

We appreciate our colleagues for their helpful suggestions.

Funding

This study was supported by The National Natural Science Foundation of China (No. 82160076), Young Talents of Yunnan Thousand Talents Plan (No. RLQB20220011), Yunnan Provincial Department of Science and Technology (No. 202301AT070093), Special Foundation Projects of Joint Applied Basic Research of Yunnan Provincial Department of Science and Technology with Kunming Medical University (No. 202201AY070001-062), 535 Talent Project of First Affiliated Hospital of Kunming Medical University (No. 2023535Q04), Hospital management project of 920th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army (2019YGC06), The Reserve Talents’ Project of Science and Technology Pioneer in Yunnan Province. (No. 2019HB045).

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  1. Chunxia Ou and Yue Zhao have contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Molecular Cardiology, Department of Cardiology, The First Affiliated Hospital of Kunming Medical University, No. 295 Xichang Rd, Kunming, 650032, Yunnan, China

    Yan Wang, Chunxia Ou, Yue Zhao, Lixing Chen, Huawei Wang, Jie Hu, Guihu Sun & Longjun Li

  2. Department of Thoracocardiac Surgery, 920th Hospital of Joint Logistics Support Force of Chinese People’s Liberation Army, No. 212 Daguan Rd, Kunming, 650032, Yunnan, China

    Jie Yu & Wenke Cai

  3. Department of Endocrinology, The First Affiliated Hospital of Kunming Medical University, No. 295 Xichang Rd, Kunming, 650032, Yunnan, China

    Shiying Huang

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Contributions

YW and JY: were responsible for the conceptualization of the project. YZ and CO: were responsible for the experimental design, application and manuscript writing. LC, WC, WW, SH, JH, GS, and LL: provided valuable suggestions for the manuscript and experiments. All authors read and approved the final manuscript.

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Correspondence to Yan Wang or Jie Yu.

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The authors have no competing interests to declare in relation to the content of this article.

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Yunnan Labreal Co., Ltd. assisted with the animal experiments and was approved by the Ethics Committee. (Issue No. PZ20211003).

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12012_2023_9818_MOESM1_ESM.tif

Supplementary file1 (TIF 43233 KB) The immunofluorescence of α-SMA and Ve-cadherin in mouse heart section, x20, scale bar: 50um, n=6.

12012_2023_9818_MOESM2_ESM.tif

Supplementary file2 (TIF 45826 KB) The immunofluorescence of α-SMA and Ve-cadherin in mouse heart section, x20, scale bar: 50um, n=6.

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Wang, Y., Yu, J., Ou, C. et al. miRNA-146a-5p Inhibits Hypoxia-Induced Myocardial Fibrosis Through EndMT. Cardiovasc Toxicol 24, 133–145 (2024). https://doi.org/10.1007/s12012-023-09818-1

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