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circCELF1 Inhibits Myocardial Fibrosis by Regulating the Expression of DKK2 Through FTO/m6A and miR-636

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Abstract

The aim of this study is to explore the role of circCELF1/miR-636/DKK2 pathway in myocardial fibrosis (MF). RT-qPCR and western blot were used to detect the expression of circCELF1, miR-636, and DKK2 in activated cardiac fibroblasts (CFs) and the hearts of acute myocardial infarction (AMI) mice. The m6A level of DKK2 was detected by RIP and RT-qPCR. The regulation of circCELF1/miR-636/DKK2 pathway on CF viability, activation, apoptosis, and migration was verified by CCK-8, western blot, flow cytometry, and Transwell. Ang II induced downregulation of circCELF1 expression, while circCELF1 enhanced the expression of DKK2 by adsorbing miR-636. circCELF1 also reduced DKK2 m6A level by upregulating FTO expression, thereby inhibiting the binding of miR-636 to DKK2 and promoting DKK2 expression. Ang II promoted CF viability, activation, and migration through the circCELF1/miR-636/DKK2 pathway. Both miR-636 inhibitors and DKK2 effectively reduced MF and improved cardiac function in AMI mice.

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Abbreviations

3′ UTR:

3′ Untranslated region

α-SMA:

α-Smooth muscle actin

AGO2:

Argonaute2

AMI:

Acute myocardial infarction

Ang II:

Angiotensin II

CFs:

Cardiac fibroblasts

circRNAs:

Circular RNAs

DKK2:

Dickkopf WNT signaling pathway inhibitor 2

FTO:

Fat mass and obesity-associated protein

HE:

Hematoxylin and Eosin

m6A:

N6-methyladenosine

METTL3:

Methyltransferase-like 3

MF:

Myocardial fibrosis

MI:

Myocardial infarction

NF-κB:

Nuclear factor kappa-B

VR:

Ventricular remodeling

RIP:

RNA immunoprecipitation

RIPA:

Radio immunoprecipitation assay buffer

RT-qPCR:

Real-time quantitative PCR

TGF-β:

Transforming growth factor-β

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Funding

This work was supported by the Shandong Provincial Medical and Health Science and Technology Development Plan Project (2019WS119) and Ningxia Medical University Scientific Research Project (XY2017113).

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Authors and Affiliations

  1. Department of Cardiology, Yantai Yuhuangding Hospital, Yantai, 264000, Shandong, China

    Xue-xun Li

  2. Department of Cardiology, General Hospital of Ningxia Medical University, Yinchuan, 750000, Ningxia, China

    Bin Mu & Xi Li

  3. Department of Cardiology, Xianyang Hospital of Yan’an University, No.38, Wenlin Road, Weicheng District, Xianyang, 712000, Shaanxi Province, People’s Republic of China

    Zi-dong Bie

Authors
  1. Xue-xun Li
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  2. Bin Mu
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  3. Xi Li
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  4. Zi-dong Bie
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Contributions

ZDB was responsible for conceiving the study. XXL was responsible for the experimental studies and manuscript preparation. BM and XL were responsible for the data acquisition and analysis.

Corresponding author

Correspondence to Zi-dong Bie.

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Ethics Approval

This study was approved by the Ethics Committee of Animal Research Institute of Xianyang Hospital of Yan'an University, and the experiment was performed in accordance with the guidelines of the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Not applicable.

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The authors declare no competing interests.

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Associate Editor Joost Sluijter oversaw the review of this article

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Li, Xx., Mu, B., Li, X. et al. circCELF1 Inhibits Myocardial Fibrosis by Regulating the Expression of DKK2 Through FTO/m6A and miR-636. J. of Cardiovasc. Trans. Res. 15, 998–1009 (2022). https://doi.org/10.1007/s12265-022-10209-0

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  • DOI: https://doi.org/10.1007/s12265-022-10209-0

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