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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Florio MC et al (2018) Aging, microRNAs and heart failure. Curr Prob Cardiol. https://doi.org/10.1016/j.cpcardiol.2018.12.003
Frolkis V et al (1991) Age-dependent effects of ischemia and reperfusion on cardiac function and Ca2+ transport in myocardium. Gerontology 37(5):233–239
Azhar G et al (1999) Ischemia-reperfusion in the adult mouse heart: influence of age☆. Exp Gerontol 34(5):699–714
Lesnefsky EJ, Hoppel CL (2003) Ischemia–reperfusion injury in the aged heart: role of mitochondria. Arch Biochem Biophys 420(2):287–297
Badalzadeh R, Yavari R, Chalabiani D (2015) Mitochondrial ATP-sensitive K+ channels mediate the antioxidative influence of diosgenin on myocardial reperfusion injury in rat hearts. Gen Physiol Biophys 34(3):323–329
Liu M et al (2012) Aging might increase myocardial ischemia/reperfusion-induced apoptosis in humans and rats. Age 34(3):621–632
Darband SG et al (2020) Combination of exercise training and L-arginine reverses aging process through suppression of oxidative stress, inflammation, and apoptosis in the rat heart. Pflügers Arch Eur J Physiol 472(2):169–178
Hosseini L, Vafaee MS, Badalzadeh R (2020) Melatonin and nicotinamide mononucleotide attenuate myocardial ischemia/reperfusion injury via modulation of mitochondrial function and hemodynamic parameters in aged rats. J Cardiovasc Pharmacol Ther 25(3):240–250
Hosseini L et al (2019) Nicotinamide adenine dinucleotide emerges as a therapeutic target in aging and ischemic conditions. Biogerontology 20(4):381–395
Poddar SK et al (2019) Nicotinamide mononucleotide: exploration of diverse therapeutic applications of a potential molecule. Biomolecules 9(1):34
Yao Z et al (2017) Nicotinamide mononucleotide inhibits JNK activation to reverse alzheimer disease. Neurosci Lett 647:133–140
Kiss T et al (2019) Nicotinamide mononucleotide (NMN) treatment attenuates oxidative stress and rescues angiogenic capacity in aged cerebromicrovascular endothelial cells: a potential mechanism for the prevention of vascular cognitive impairment. GeroScience 41(5):619–630
Dominguez-Rodriguez A et al (2010) Melatonin and circadian biology in human cardiovascular disease. J Pineal Res 49(1):14–22
Jiki Z, Lecour S, Nduhirabandi F (2018) Cardiovascular benefits of dietary melatonin: a myth or a reality? Front Physiol 9:528
Zhang Y et al (2019) Melatonin attenuates myocardial ischemia-reperfusion injury via improving mitochondrial fusion/mitophagy and activating the AMPK-OPA1 signaling pathways. J Pineal Res 66(2):e12542
Zhang B et al (2013) SIRT3 overexpression antagonizes high glucose accelerated cellular senescence in human diploid fibroblasts via the SIRT3–FOXO1 signaling pathway. Age 35(6):2237–2253
Porter GA et al (2014) SIRT3 deficiency exacerbates ischemia-reperfusion injury: implication for aged hearts. Am J Physiol Heart Circ Physiol 306(12):H1602–H1609
Sengupta A et al (2011) FoxO transcription factors promote cardiomyocyte survival upon induction of oxidative stress. J Biol Chem 286(9):7468–7478
Juhasz B et al (2008) Bromelain induces cardioprotection against ischemia-reperfusion injury through Akt/FOXO pathway in rat myocardium. Am J Physiol Heart Circ Physiol 294(3):H1365–H1370
Klimova N, Long A, Kristian T (2019) Nicotinamide mononucleotide alters mitochondrial dynamics by SIRT3-dependent mechanism in male mice. J Neurosci Res 97(8):975–990
Yamamoto T et al (2014) Nicotinamide mononucleotide, an intermediate of NAD+ synthesis, protects the heart from ischemia and reperfusion. PLoS ONE 9(6):e98972
Bayrami G et al (2018) Effect of ischemic postconditioning on myocardial function and infarct size following reperfusion injury in diabetic rats pretreated with vildagliptin. J Cardiovasc Pharmacol Ther 23(2):174–183
Xin W et al (2011) Involvement of endoplasmic reticulum stress-associated apoptosis in a heart failure model induced by chronic myocardial ischemia. Int J Mol Med 27(4):503–509
Tao L et al (2019) Retinol palmitate protects against myocardial ischemia/reperfusion injury via reducing oxidative stress and inhibiting apoptosis. Am J Transl Res 11(3):1510
Chakrabarti S, Hoque AE, Karmazyn M (1997) A rapid ischemia-induced apoptosis in isolated rat hearts and its attenuation by the sodium–hydrogen exchange inhibitor HOE 642 (cariporide). J Mol Cell Cardiol 29(11):3169–3174
Zhang Y et al (2016) Exogenous NAD+ administration significantly protects against myocardial ischemia/reperfusion injury in rat model. Am J Transl Res 8(8):3342
Dobsak P et al (2003) Melatonin protects against ischemia-reperfusion injury and inhibits apoptosis in isolated working rat heart. Pathophysiology 9(3):179–187
Hosseini L et al (2019) Nicotinamide mononucleotide and melatonin alleviate aging-induced cognitive impairment via modulation of mitochondrial function and apoptosis in the prefrontal cortex and hippocampus. Neuroscience 423:29–37
Meng H et al (2019) SIRT3 regulation of mitochondrial quality control in neurodegenerative diseases. Front Aging Neurosci 11:313
Zhai M et al (2017) Melatonin ameliorates myocardial ischemia reperfusion injury through SIRT 3-dependent regulation of oxidative stress and apoptosis. J Pineal Res 63(2):e12419
Ping L, Yin Z, Li Y (2016) GW27-e0632 NR reduced myocardial ischemia-reperfusion injury by improving mitochondrial biogenesis and reducing excessive autophagy via Sirt3-PGC-1α/P53 pathway. J Am Coll Cardiol 68(16 Supplement):C24–C25
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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The authors declare that they have no conflicts of interest to disclose.
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). https://doi.org/10.1007/s11033-021-06351-8