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Cardioprotective effects of chloroquine pretreatment on ischemic and reperfusion injury via activation of ERK1/2 in isolated rat hearts

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Abstract

Purpose

Several therapeutic agents have been found to prevent myocardial ischemic and reperfusion (I/R) injury after cardiac surgery; however, no drug is routinely used to afford cardioprotective benefits in clinical settings. Herein, we aimed to determine whether chloroquine (CQ) pretreatment attenuates I/R injury after global ischemia in isolated rat hearts and elucidate mechanisms underlying the effects of CQ.

Methods

Isolated rat hearts were subjected to 30-min global ischemia, followed by 60-min reperfusion with Krebs–Henseleit buffer (KHB). Immediately before ischemia, 10 mL of pretreatment solutions (KHB, n = 4 or KHB + CQ [100 μM], n = 4) were injected through the aortic root. Cardiac function was examined based on the rate pressure product (RPP). Myocardial apoptosis was evaluated using TUNEL staining. To assess the reperfusion ischemia salvage kinase pathway, protein expression levels of AKT and extracellular signal-regulated kinase (ERK1/2) were determined using western blotting. To investigate the role of ERK1/2, an ERK1/2 selective inhibitor was used in eight additional rats.

Results

The recovery rate of the RPP was higher in the KHB + CQ group than in the KHB group 60 min after I/R (KHB, 44 ± 3% vs. KHB + CQ, 69 ± 7%; P = 0.019, d = 2.2). CQ pretreatment reduced apoptosis and enhanced the phosphorylation of ERK1/2; however, AKT phosphorylation was unaltered. In addition, the ERK1/2 inhibitor abolished CQ-mediated cardioprotective effects.

Conclusions

CQ pretreatment showed protective effects on cardiac function after I/R by activating ERK1/2.

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Data availability

The data sets generated during the current study will be shared on reasonable requests to the corresponding author.

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Acknowledgements

The authors thank Editage for the English language editing. We also thank the Sapporo General Pathology Laboratory and Ryota Azuma for technical assistance with the experiments.

Funding

The authors have no funds, Grants, or other support.

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Authors and Affiliations

  1. Department of Cardiovascular and Thoracic Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kitaku, Sapporo, 060-8638, Japan

    Ryota Murase, Yasushige Shingu & Satoru Wakasa

Authors
  1. Ryota Murase
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  2. Yasushige Shingu
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  3. Satoru Wakasa
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by RM and YS. Manuscript revision was performed by SW. The first draft of the manuscript was written by RM and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Yasushige Shingu.

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Competing of interests

The authors declare that they have no conflicts of interest.

Ethical approval

All procedures were approved by the National University Corporation Hokkaido University Animal Research Committee and consistent with the Guide for the Care and Use of Laboratory Animals, published by the US National Institute of Health (NIH publication No. 85-23, revised 1996).

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11033_2022_7801_MOESM3_ESM.tif

Supplementary file3 (TIF 8053 KB) Fig. 1 Recovery rate of RPP with different concentrations of CQ. Values are mean ± SEM. N = 4 for each group. * P < 0.05, ** P < 0.01. CQ, chloroquine; KHB, Krebs-Henseleit buffer; RPP, rate pressure product

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Murase, R., Shingu, Y. & Wakasa, S. Cardioprotective effects of chloroquine pretreatment on ischemic and reperfusion injury via activation of ERK1/2 in isolated rat hearts. Mol Biol Rep 49, 9429–9436 (2022). https://doi.org/10.1007/s11033-022-07801-7

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  • DOI: https://doi.org/10.1007/s11033-022-07801-7

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