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MicroRNAs sequencing of plasma exosomes derived from patients with atrial fibrillation: miR-124-3p promotes cardiac fibroblast activation and proliferation by regulating AXIN1

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Abstract

MicroRNAs (miRNAs) play an important role in the pathogenesis of atrial fibrillation (AF). Exosomal miRNAs may develop as promising biomarkers for AF. To explore significant exosomal miRNAs in AF, plasma exosomes were extracted from 3 patients with AF and 3 patients with sinus rhythm (SR), respectively. Differential expression of exosomal miRNAs were screened by high-throughput sequencing analysis and verified by qRT-PCR from 40 patients with AF and 40 patients with SR. The target genes prediction, biological function, and signaling pathways analysis were conducted by miRanda software, gene ontology (GO), and KEGG analysis. The results showed that there were 40 differently expressed exosomal miRNAs from AF patients compared with SR patients, of which 13 miRNAs were upregulated and 27 miRNAs were downregulated. qRT-PCR validation demonstrated that miR-124-3p, miR-378d, miR-2110, and miR-3180-3p were remarkably upregulated, while miR-223-5p, miR-574-3p, miR-125a-3p, and miR-1299 were downregulated. To explore the function of miR-124-3p associated with AF, plasma exosomes derived from AF patients were co-incubated with rat myocardial fibroblasts. The expression of miR-124-3p was upregulated in myocardial fibroblasts. The viability and proliferation of myocardial fibroblasts were elevated by transfecting with miR-124-3p overexpression plasmids using CCK8 and immunofluorescence-staining methods. AXIN1 was verified to be the target of miR-124-3p by luciferase assay in vitro. Expression of AXIN1 was reduced, while β-catenin, Collagen 1, and α-SMA were increased in myocardial fibroblasts with miR-124-3p overexpression. In conclusion, these findings suggested that circulating exosomal miRNAs may serve as novel biomarkers for AF, and miR-124-3p promotes fibroblast activation and proliferation through regulating WNT/β-catenin signaling pathway via AXIN1.

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Data availability

The data used to support the findings of this study are available from the corresponding author upon request.

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Funding

This work was supported by the National Natural Science Foundation of China (grant numbers 81770334, 81970281); Taishan Scholar Engineering Construction Fund of Shandong Province (grant number ts201511104); Academic promotion programme of Shandong First Medical University (grant number 2019QL012); and Natural Science Foundation of Shandong Province (grant number ZR2020QH014).

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Authors and Affiliations

  1. Department of Cardiology, Shandong Provincial Qianfoshan Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China

    Pengju Zhu, Huilin Li, An Zhang & Yinglong Hou

  2. Department of Cardiology, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Shandong Medicine and Health Key Laboratory of Cardiac Electrophysiology and Arrhythmia, Jinan, China

    Pengju Zhu, Zhan Li, Yong Zhang, Manyi Ren, Yujiao Zhang & Yinglong Hou

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  1. Pengju Zhu
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Contributions

All authors contributed to the study conception and design. Experimental funding acquisition, study guidance, and supervision were performed by Yujiao Zhang and Yinglong Hou. Material preparation, data collection, and analysis were performed by Huilin Li, An Zhang, Zhan Li, Yong Zhang, and Manyi Ren. The first draft of the manuscript was written by Pengju Zhu. All authors commented on previous versions of the manuscript and approved the final manuscript.

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Correspondence to Yujiao Zhang or Yinglong Hou.

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This study was approved by the Ethics Committee of Shandong University (Approval No. 2018S0021).

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Key points

There was differential expression of exosomal miRNAs between the patients with AF and SR, in which miR-124-3p was significantly overexpressed in AF.

MiR-124-3p promotes rat cardiac fibroblast activation and proliferation by regulating WNT/β-catenin pathway via AXIN1.

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Zhu, P., Li, H., Zhang, A. et al. MicroRNAs sequencing of plasma exosomes derived from patients with atrial fibrillation: miR-124-3p promotes cardiac fibroblast activation and proliferation by regulating AXIN1. J Physiol Biochem 78, 85–98 (2022). https://doi.org/10.1007/s13105-021-00842-9

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  • DOI: https://doi.org/10.1007/s13105-021-00842-9

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