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Mitochondrial Oxidative Phosphorylation Function and Mitophagy in Ischaemic/Reperfused Hearts from Control and High-Fat Diet Rats: Effects of Long-Term Melatonin Treatment

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Abstract

Purpose

Oxidative stress causes mitochondrial dysfunction in myocardial ischaemia/reperfusion (I/R) as well as in obesity. Mitochondrial depolarization triggers mitophagy to degrade damaged mitochondria, a process important for quality control. The aims of this study were to evaluate (i) the effect of I/R on mitochondrial oxidative phosphorylation and its temporal relationship with mitophagy in hearts from obese rats and their age-matched controls, and (ii) the role of oxidative stress in these processes using melatonin, a free radical scavenger.

Methods

Male Wistar rats were divided into 4 groups: control (normal diet ± melatonin) and high-fat sucrose diet (HFSD ± melatonin). Rats received melatonin orally (10 mg/kg/day). After 16 weeks, hearts were removed and subjected to 40-min stabilization, and 25-min global ischaemia/10-min reperfusion for preparation of mitochondria. Mitochondrial oxidative phosphorylation was measured polarographically. Western blotting was used for evaluation of PINK1, Parkin, p62/SQSTM1 (p62) and TOM 70. Infarct size was measured using tetrazolium staining.

Results

Ischaemia and reperfusion respectively reduced and increased mitochondrial QO2 (state 3) and the ox-phos rate in both control and HFSD mitochondria, showing no major changes between the groups, while melatonin pretreatment had little effect. p62 as indicator of mitophagic flux showed up- and downregulation of mitophagy by ischaemia and reperfusion respectively, with melatonin having no significant effect. Melatonin treatment caused a significant reduction in infarct size in hearts from both control and diet groups.

Conclusions

The results suggest that I/R (i) affects mitochondria from control and HFSD hearts similarly and (ii) melatonin-induced cardioprotection is not associated with reversal of mitochondrial dysfunction or changes in the PINK1/Parkin pathway.

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Funding

The project was funded by the South African National Research Foundation, Grant no 93579.

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Authors and Affiliations

  1. Division of Medical Physiology, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, University of Stellenbosch, PO Box 241, Cape Town, 8000, South Africa

    Karthik Dhanabalan, Sibonginkosi Mzezewa, Barbara Huisamen & Amanda Lochner

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  1. Karthik Dhanabalan
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  2. Sibonginkosi Mzezewa
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  3. Barbara Huisamen
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  4. Amanda Lochner
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Correspondence to Amanda Lochner.

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Ethical Approval

Male Wistar rats were used as experimental animals. The breeding, maintenance and feeding of these rats as well as food and water consumption, and weight monitoring were all carried out in the Central Research Facility, Faculty of Health Sciences, Stellenbosch University The study conformed to the revised South African National Standard for the Care and Use of Animals for Scientific Purposes (South African Bureau of Standards, SANS 10386, 2008) and was approved by the Committee for the ethical use of animals in research of the University of Stellenbosch (project number SU-ACUM14-00039).

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Dhanabalan, K., Mzezewa, S., Huisamen, B. et al. Mitochondrial Oxidative Phosphorylation Function and Mitophagy in Ischaemic/Reperfused Hearts from Control and High-Fat Diet Rats: Effects of Long-Term Melatonin Treatment. Cardiovasc Drugs Ther 34, 799–811 (2020). https://doi.org/10.1007/s10557-020-06997-9

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