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Nicotinamide mononucleotide and melatonin counteract myocardial ischemia-reperfusion injury by activating SIRT3/FOXO1 and reducing apoptosis in aged male rats

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It has been documented that aging increases the risk of cardiovascular disease including myocardial ischemia/reperfusion (IR) injury and acute myocardial infarction. In this study, we aimed to investigate the individual or combined effects of nicotinamide mononucleotide (NMN) and melatonin (Mel) treatment on apoptotic markers, expression of SIRT3, and FOXO1, and infarct size of the aged myocardium subjected to IR injury. Sixty aged Wistar rats (22–24 months) were assigned to five groups including sham, IR, NMN+IR, Mel+IR, and NMN+Mel+IR (combination therapy). Isolated hearts were exposed to 30-min regional ischemia followed by 60-min reperfusion. NMN (100 mg/kg/day/i.p.) was injected every second day starting on day 28 before IR injury. Melatonin was added to the perfusion solution five minutes prior to and until 15 min after the start of reperfusion. The infarct size was assessed by computerized planimetry. The mRNA levels of SIRT3, FOXO1, and apoptotic genes Bax, Bcl-2, and Caspase-3 were estimated by real-time PCR. All treatments reduced infarct size as compared with the IR group. Melatonin and NMN upregulated the gene expression of Bcl‐2, SIRT3, and FOXO1 and downregulated the gene expression of Bax, and Caspase‐3, in comparison to the IR group. Also, the protein levels of SIRT3, quantified by Western blotting, were upregulated by the interventions. The effects of combination therapy were significantly greater than those of melatonin or NMN alone. These findings indicate that the combined administration of NMN and melatonin can protect the aged heart against IR injury by decreasing apoptosis and activating the SIRT3/FOXO1 pathway.

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This study was derived from the thesis of A Jafari-Azad and supported by a grant from Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz-Iran.


This work was supported by a Grant (59024) from Drug Applied Research Center, and Molecular Medicine Research Center, Tabriz University of Medical Sciences, Tabriz-Iran.

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AJ, MV, and RB conceived and designed research. AJ, MR, and LH conducted experiments. AJ, SF, and PH contributed new reagents or analytical tools. AJ and LH analyzed data. AJ, LH, MR, and PH wrote and critically revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Reza Badalzadeh.

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The authors declare that they have no conflicts of interest to disclose.

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All steps of this study were performed under the control of local animal research committee according to the standard guidelines.

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Jafari-Azad, A., Hosseini, L., Rajabi, M. et al. Nicotinamide mononucleotide and melatonin counteract myocardial ischemia-reperfusion injury by activating SIRT3/FOXO1 and reducing apoptosis in aged male rats. Mol Biol Rep 48, 3089–3096 (2021).

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