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Down–regulating miR-217-5p Protects Cardiomyocytes against Ischemia/Reperfusion Injury by Restoring Mitochondrial Function via Targeting SIRT1

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Abstract

Downregulating miR-217-5p could protect cardiomyocytes against ischemia/reperfusion (I/R) injury, but its role in restoring mitochondrial function of I/R-injured cardiomyocytes remained unclear. H9C2 cardiomyocyte-derived cell line with I/R injury was established in vitro on the basis of hypoxia/reperfusion (H/R) model. Cell viability and apoptosis were respectively detected by MTT assay and flow cytometry. Contents of lactate dehydrogenase (LDH) and adenosine triphosphate (ATP) were determined. Flow cytometry was performed to measure the production of reactive oxygen species (ROS) and mitochondrial membrane potential (MMP). Target gene and potential binding sites between miR-217-5p and Sirtuin1 (SIRT1) were predicted by TargetScan and confirmed by dual-luciferase reporter assay. Relative SIRT1 and expressions of autophagy-related and apoptosis-related genes were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. After I/R treatment, the viability of H9C2 cardiomyocyte-derived cell line and ATP contents were reduced, but LDH and ROS contents were increased, at the same time, cell apoptosis and the expressions of miR-217-5p, p62 and cleaved caspase-3 were increased, whereas the expressions of SIRT1, LC3 (light chain 3), PINK1 (PTEN-induced kinase 1), Parkin, Bcl-2, and c-IAP (inhibitor of apoptosis protein) were reduced. However, downregulating miR-217-5p expression reversed the effects of I/R. SIRT1 was predicted and verified to be the target of miR-217-5p, and silencing SIRT1 reversed the effects of downregulating miR-217-5p on I/R-injured cells. Downregulating miR-217-5p could help restore mitochondrial function via targeting SIRT1, so as to protect cardiomyocytes against I/R-induced injury.

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Data Availability

The analyzed data sets generated during the study are available from the corresponding author on reasonable request.

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Funding

This work was financially supported by the Science and Technology Foundation of Tianjin Health Bureau (Grant Number 2015ky31).

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  1. Yujuan Qi and Kai Zhang contributed equally to this work.

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  1. ICU, Tianjin Chest Hospital, No. 261, South Tai’erzhuang Road, Jinnan District, Tianjin, 300000, China

    Yujuan Qi, Kai Zhang, Peijun Li & Zhenhua Wu

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YQ designed the research study. YQ and PL performed the research. PL and ZW analyzed the data. YQ wrote the manuscript. All authors contributed to editorial changes in the manuscript. All authors read and approved the final manuscript.

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Correspondence to Peijun Li.

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Qi, Y., Zhang, K., Li, P. et al. Down–regulating miR-217-5p Protects Cardiomyocytes against Ischemia/Reperfusion Injury by Restoring Mitochondrial Function via Targeting SIRT1. Inflammation 44, 383–396 (2021). https://doi.org/10.1007/s10753-020-01343-5

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