Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Neurosciences Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Mol Biol Rep. 2021 Apr;48(4):3089-3096. doi: 10.1007/s11033-021-06351-8. Epub 2021 Apr 17.
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.
已有文献证明，衰老会增加心血管疾病的风险，包括心肌缺血/再灌注（IR）损伤和急性心肌梗死。在这项研究中，我们旨在研究烟酰胺单核苷酸（NMN）和褪黑素（Mel）单独或联合治疗对IR 损伤老龄心肌细胞凋亡标志物、SIRT3 和 FOXO1 表达以及梗死面积的影响。60 只老龄 Wistar 大鼠（22-24 个月）分为 5 组，包括假手术组、IR 组、NMN+IR 组、Mel+IR 组和 NMN+Mel+IR（联合治疗组）。离体心脏先暴露于 30min 局部缺血，然后再灌注 60min。NMN（100mg/kg/天，腹腔注射）从 IR 损伤前 28 天开始，每隔一天注射一次。Melatonin 在再灌注开始前 5 分钟加入灌注液中，并持续到再灌注 15 分钟。通过计算机定积分评估梗死面积。通过实时 PCR 估计 SIRT3、FOXO1 和凋亡基因 Bax、Bcl-2 和 Caspase-3 的 mRNA 水平。与 IR 组相比，所有治疗均减少了梗死面积。与 IR 组相比，褪黑素和 NMN 上调了 Bcl-2、SIRT3 和 FOXO1 的基因表达，下调了 Bax 和 Caspase-3 的基因表达。此外，通过 Western 印迹法量化的 SIRT3 蛋白水平也因干预而升高。联合治疗的效果明显大于单独使用褪黑素或 NMN。这些发现表明，NMN 和褪黑素联合给药可以通过减少细胞凋亡和激活 SIRT3/FOXO1 通路来保护老龄心脏免受 IR 损伤。
