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Modulation of autophagy as the target of mesenchymal stem cells-derived conditioned medium in rat model of myocardial ischemia/reperfusion injury

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Abstract

Human amniotic membrane mesenchymal stem cells-derived conditioned medium (hAM-MSCs-CM) has positive effects against myocardial ischemia/reperfusion (MI/R) injury. However, it needs further investigations how hAM-MSCs-CM leads to the cell survival under MI/R via modulation of autophagy. The purpose of this study is investigating the effects of hAM-MSCs-CM in a rat model of MI/R injury by focusing on the role of autophagy as one of its possible mechanisms. Male Wistar rats (44 rats, 175–200 g) were randomly divided into four groups: Sham, MI/R, culture media-receiving and conditioned medium-receiving. MI/R was induced by 30 min of left anterior descending coronary artery ligation. After 15 min reperfusion, culture media or hAM-MSCs-CM (150 μl) were injected intramyocardially. At the end of the experiment, CK-MB, autophagy markers, phosphorylated and total forms of mTOR and ULK1, cardiac function and fibrosis were measured. hAM-MSCs-CM significantly decreased CK-MB levels (P < 0.0001), and also the mRNA levels of Beclin1 (P < 0.0001), LC3 (P = 0.012) and p62 (P = 0.003). In addition, hAM-MSCs-CM significantly reduced Beclin1, LC3II/LC3I and p62 protein levels (P < 0.0001), and increased p-mTOR/mTOR (P = 0.022) and p-ULK1/ULK1 (P < 0.0001) expressions. Moreover, hAM-MSCs-CM improved cardiac function and decreased fibrosis (P < 0.0001). This study showed cardioprotective effects of hAM-MSCs-CM against MI/R injury through modulation of autophagy via mTOR/ULK1 pathway. Based on these findings, it can be concluded that hAM-MSCs-CM can be offered as an attractive candidate for attenuation of MI/R injury in future, but needs further investigations.

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Acknowledgements

This study was part of a Ph.D. thesis of Behnaz Mokhtari and financially supported by Physiology Research Center in Iran University of Medical Sciences, Tehran, Iran.

Funding

The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work has been supported by a grant (Grant Number: 97-2-3-33054) from Iran University of Medical Sciences, Tehran-Iran.

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Authors and Affiliations

  1. Physiology Research Center, Iran University of Medical Sciences, Tehran, Islamic Republic of Iran

    Behnaz Mokhtari & Nahid Aboutaleb

  2. Department of Physiology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Islamic Republic of Iran

    Behnaz Mokhtari & Nahid Aboutaleb

  3. Molecular Medicine Research Center, Tabriz University of Medical Sciences, Tabriz, Islamic Republic of Iran

    Reza Badalzadeh

  4. Department of Physiology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Islamic Republic of Iran

    Reza Badalzadeh

Authors
  1. Behnaz Mokhtari
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  2. Reza Badalzadeh
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  3. Nahid Aboutaleb
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Contributions

BM and NA did the study design. BM performed experimental tests, gathered and analyzed the data, and drafted the manuscript. RB contributed in interpretation of the results and finalized the manuscript editing and critically revised the manuscript. NA supervised the whole project and revised the manuscript. All gave final approval and agree to be accountable for all aspects of work ensuring integrity and accuracy.

Corresponding author

Correspondence to Nahid Aboutaleb.

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Conflict of interest

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Ethical approval

Animal care procedures, as well as all experimental protocols were approved by the Institutional Animal Ethical Committee of Iran University of Medical Sciences (Ethical code: IR.IUMS.FMD.REC.1397.234).

Informed consent

Amniotic membranes were supplied from Shahid Akbar Abadi Hospital under informed agreement from each participants in accordance with ethical standards of the institutional and/or national research committee and with the 1975 Declaration of Helsinki as reflected in a prior approval by the institution's human research committee.

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Mokhtari, B., Badalzadeh, R. & Aboutaleb, N. Modulation of autophagy as the target of mesenchymal stem cells-derived conditioned medium in rat model of myocardial ischemia/reperfusion injury. Mol Biol Rep 48, 3337–3348 (2021). https://doi.org/10.1007/s11033-021-06359-0

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  • DOI: https://doi.org/10.1007/s11033-021-06359-0

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