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MiR-21 participates in LPS-induced myocardial injury by targeting Bcl-2 and CDK6

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Abstract

Objective

This study aimed to investigate the relationship between miR-21 and lipopolysaccharide (LPS)-induced myocardial injury and its molecular and regulatory mechanisms.

Methods

We constructed LPS-mediated myocardial injury model using C57BL/6J mice and H9c2 cells. In-vivo, in-vitro, RIP and dual-luciferase reporter assays were used to determine the effect of miR-21 on myocardial injury.

Results

In-vivo and in-vitro results showed that the expression of miR-21 was increased in LPS-treated H9c2 cells and myocardial tissues of mice, and the pro-inflammatory cytokines (IL-1β, IL-6, IL-8 and TNF-α) and NF-κB pathway were activated in LPS-treated H9c2 cells. Besides, the B-cell lymphoma-2 (Bcl-2) and cyclin-dependent kinase 6 (CDK6) expression levels decreased, while Bax and cleaved caspase 9 levels increased in LPS-treated H9c2 cells. Inhibition of miR-21 could suppress LPS-induced apoptosis, inflammatory reactions and NF-κB activation to attenuate LPS-induced myocardial injury in H9c2 cells, and effectively improve survival of mice with sepsis. Most importantly, Bcl-2 and CDK6 were found to be the direct target of miR-21 using dual-luciferase reporter and RNA immunoprecipitation assays. Further gain-of-function assay demonstrated that Bcl-2 or CDK6 over-expression promoted the protective effects of miR-21 inhibitor on LPS-mediated myocardial cells.

Conclusion

Our findings revealed that the down-regulation or antagonism of miR-21 protects myocardial cells against LPS-induced apoptosis and inflammation through up-regulating Bcl-2 and CDK6 expression, which provided a new insight for prevention and treatment of myocardial injury.

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Abbreviations

Bcl-2:

B-cell lymphoma 2

CDK6:

Cyclin-dependent kinase 6

ELISA:

Enzyme-linked immunosorbent assay

LPS:

Lipopolysaccharide

miR-21:

MiRNA-21

PBS:

Phosphate-buffered saline

RT-qPCR:

Real-time quantitative PCR

RIP:

Immunoprecipitation

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Authors and Affiliations

  1. Department of Cardiovascular Medicine, Weifang No. 2 People’s Hospital, Weifang, Shandong, People’s Republic of China

    Yu Li

  2. Department of Intensive Care Unit, Sunshine Union Hospital, Weifang, Shandong, People’s Republic of China

    Guanghui Sun

  3. Department of Cardiovascular Medicine, Sunshine Union Hospital, No. 9000, Yingqian Street, Gaoxin District, Weifang, Shandong, People’s Republic of China

    Luqiang Wang

Authors
  1. Yu Li
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  2. Guanghui Sun
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  3. Luqiang Wang
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Correspondence to Luqiang Wang.

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The authors declare that they have no conflict of interest.

Ethical approval

All experimental animal procedures in this study conformed to the National Institutes of Health Guide for the Care and Use of Laboratory Animals, and were approved by the Institutional Animal Care and Use Committee of Sunshine Union Hospital.

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Yes from all authors.

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Responsible Editor: John Di Battista.

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Li, Y., Sun, G. & Wang, L. MiR-21 participates in LPS-induced myocardial injury by targeting Bcl-2 and CDK6. Inflamm. Res. 71, 205–214 (2022). https://doi.org/10.1007/s00011-021-01535-1

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  • DOI: https://doi.org/10.1007/s00011-021-01535-1

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