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Inhibition of miR-217 Protects Against Myocardial Ischemia–Reperfusion Injury Through Inactivating NF-κB and MAPK Pathways

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Abstract

Purpose

Recent studies have demonstrated that miRNAs play a vital role in regulating myocardial ischemia/reperfusion injury (MIRI). MiR-217 has been proven to be implicated in cardiac diseases such as chronic heart failure and cardiac myxoma. However, the role of miR-217 in MIRI is not clear.

Methods

A mouse MIRI model was established and the myocardial infarct size was evaluated by TTC staining. The expression level of miR-217 in I/R group was determined by real-time polymerase chain reaction. Subsequently, MIRI mice and H9C2 cells were administrated with miR-217 inhibitor in vivo and in vitro, respectively. The levels of TNF-α and IL-6 were measured by commercially available ELISA kits. Blood and cell samples were collected for the measurement of lactate dehydrogenase (LDH) level and caspase-3 activity. Cell viability was assessed with the CCK-8 assay. We then explored the detailed molecular mechanisms by TargetScan 7.1 database and further studies were performed to prove the prediction by dual-luciferase reporter assay.

Results

Larger stainless infarct areas were observed in the MIRI group, accompanied by inceased serum LDH activity, indicating the mouse MIRI model was successfully established. MiR-217 was up-regulated in MIRI mice and hypoxia/reoxygenation-treated H9C2 cells. MiR-217 knockdown alleviated the MIRI in MIRI mouse model, and also attenuated the myocardial hypoxia/reoxygenation injury in H9C2 cells. Moreover, dual specificity protein phosphatase 14 (DUSP14) was proved to be a target of miR-217. Besides, further study indicated that inhibition of miR-217 protected against MIRI through inactivating NF-κB and MAPK pathways via targeting DUSP14.

Conclusions

MiR-217 inhibition protected against MIRI through inactivating NF-κB and MAPK pathways by targeting DUSP14. This study may provide valuable diagnostic and factors and therapeutic agents for MIRI.

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Data Availability

All data generated or analyzed during this study are included in this published article.

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Conflict of interest

The authors declare that they have no competing interests, and all authors should confirm its accuracy.

Ethics Approval

The animal use protocol listed below has been reviewed and approved by the Animal Ethical and Welfare Committee.

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Authors and Affiliations

  1. Department of Cardiovascular Medicine, Heji Hospital Affiliated to Changzhi Medical College, 271 East Taihang Street, Changzhi, 046000, Shanxi, China

    Yanfang Li, Liping Fei, Junli Wang & Qingying Niu

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  1. Yanfang Li
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Contributions

YFL conceived and designed the experiments, LPF analyzed and interpreted the results of the experiments, JLW and QYN performed the experiments.

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Correspondence to Yanfang Li.

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Associate Editor Hanjoong Jo oversaw the review of this article.

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Li, Y., Fei, L., Wang, J. et al. Inhibition of miR-217 Protects Against Myocardial Ischemia–Reperfusion Injury Through Inactivating NF-κB and MAPK Pathways. Cardiovasc Eng Tech 11, 219–227 (2020). https://doi.org/10.1007/s13239-019-00452-z

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