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miR-451-3p alleviates myocardial ischemia/reperfusion injury by inhibiting MAP1LC3B-mediated autophagy

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Abstract

Objective and design

We aim to explore the molecular mechanism of myocardial ischemia–reperfusion injury (MIRI).

Methods

The H9C2 cells were cultured under hypoxia/reoxygenation (H/R) condition to induce myocardial injury in vitro. The expression of miR-451-3p and MAP1LC3B was detected by RT-qPCR. Dual-luciferase reporter assay and RNA pull-down assay were performed to examine the relationship between microRNA (miR)-451-3p and MAP1LC3B. CCK8 was used to test cell viability. The level of LDH and CK was evaluated via ELISA. Immunofluorescence assay and flow cytometry were applied to detect autophagy and apoptosis, respectively. Autophagy-related protein expressions were determined by western blotting. Furthermore, an in vivo rat model of MIRI was established by subjection to 30 min ischemia and subsequently 24 h reperfusion for validation of the role of miR-451-3p in regulating MIRI in vivo.

Results

miR-451-3p was down-regulated in MIRI, and miR-451-3p mimics transfection alleviated autophagy and apoptosis induced by MIRI. miR-451-3p could target MAP1LC3B directly. Co-transfection miR-451-3p mimics and pcDNA 3.1 MAP1LC3B curbed the protected effects of miR-451-3p mimics on MIRI.

Conclusions

miR-451-3p played a protective role in MIRI via inhibiting MAP1LC3B-mediated autophagy, which may provide new molecular targets for the treatment of MIRI and further improves the clinical outcomes of heart diseases.

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Data availability

All data generated or analyzed during this study are included in this article. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

MIRI:

Myocardial ischemia–reperfusion injury

H/R:

Hypoxia/reoxygenation

AMI:

Acute myocardial infarction

MAP1LC3/ LC3:

Microtubule-associated protein 1 light chain 3

FITC:

Fluorescein isothiocyanate

MAP1LC3B-WT:

MAP1LC3B wild-type

3′-UTR:

3′-Untranslated region

MAP1LC3B –MUT:

MAP1LC3B mutated-type

LAD:

The left anterior descending coronary artery

AI:

Apoptosis index

ANOVA:

Analysis of variance

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Acknowledgements

We would like to give our sincere gratitude to the reviewers for their constructive comments. This work was supported by Natural Science Foundation of Guangxi (No.2018GXNSFDA281039, No.2018GXNSFAA050110, No. 2020GXNSFAA297009).

Funding

This work was supported by Natural Science Foundation of Guangxi (No.2018GXNSFDA281039, No.2018GXNSFAA050110, No. 2020GXNSFAA297009).

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  1. Xiang-Wei Lv and Zi-Feng He are co-first authors.

Authors and Affiliations

  1. Department of Cardiology, Affiliated Hospital of Guilin Medical University, No.20, Lequn Road, Guilin, 541001, Guangxi Zhuang Autonomous Region, People’s Republic of China

    Xiang-Wei Lv, Zi-Feng He, Pan-Pan Zhu, Qiu-Yu Qin, Yun-Xue Han & Tong-Tong Xu

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Contributions

Conception and study design: XWL. Data acquisition: ZFH, PPZ. Data analysis: QYQ. Manuscript drafting: YXH. Manuscript revising: TTX. All authors have read and approved the final version of this manuscript to be published.

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Correspondence to Tong-Tong Xu.

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Lv, XW., He, ZF., Zhu, PP. et al. miR-451-3p alleviates myocardial ischemia/reperfusion injury by inhibiting MAP1LC3B-mediated autophagy. Inflamm. Res. 70, 1089–1100 (2021). https://doi.org/10.1007/s00011-021-01508-4

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