Cardioprotective effect of 3-nitro-N-methyl salicylamide on the isolated rat heart under cold ischemia preservation

doi:10.12307/2022.223

Abstract

Abstract: BACKGROUND: During reperfusion, increased reactive oxygen species will cause oxidative stress injury in ischemic organs, which is the critical factor for ischemia and reperfusion injury. Cellular reactive oxygen species are mainly originated from mitochondrial electron transport chain. Therefore, it is an effective strategy to reduce myocardial ischemia/reperfusion injury due to elevated reactive oxygen species by inhibiting electron transport at mitochondrial electron transport chain. 3-Nitro-N-methyl-salicylamide (3-NNMS) is a semi-inhibitor for electron transport chain complex III, which can slow down the electron transport rate. 3-NNMS has potential applications for cardiac preservation; nevertheless, no related definite research or clinical practice has been reported.
OBJECTIVE: To investigate the protective effect of a new cardioplegic solution with 3-NNMS as the main component on isolated rat heart preservation for 8 hours.
METHODS: Heart specimens were taken from healthy male Wistar rats, perfused stably for 30 minutes, and then preserved in different cardioplegic solutions at low temperature for 8 hours. According to different cardioplegic solutions used, rat heart specimens were divided into a control group (no preservation), a 3-NNMS cardioplegic solution group, a Celsior cardioplegic group, and a 3-NNMS+Celsior cardioplegic solution group. The hemodynamic changes of the heart were detected by Powerlab instruments. The mitochondrial function in the preserved myocardium was measured by Oxygraph-2k High-resolution respirometry and chemiluminescence apparatus. Myocardial injury was assessed by detecting the expression of cardiac troponin T, creatine kinase isoenzyme MB, and lactate dehydrogenase using ELISA. The morphological changes of the heart were histologically observed. And the myocardial reactive oxygen species level was tested by inverted fluorescence microscope.
RESULTS AND CONCLUSION: The 3-NNMS cardioplegic solution could improve the heart rate recovery compared with the Celsior (P < 0.05), and decrease the levels of cardiac troponin T, creatine kinase isoenzyme MB, and lactate dehydrogenase in perfusate supernatant (P < 0.05). 3-NNMS could remarkably elevate mitochondria membrane potential, and maintain mitochondrial membrane structure effectively (P < 0.05). 3-NNMS could significantly increase the activity of superoxide dismutase (P < 0.05), ameliorate myocardial antioxidant function, and alleviate oxidative damage. It was nontoxic for cell culture with 3-NNMS, which can be used regardless of the concentration. To conclude, 3-NNMS can reduce oxidative stress-induced myocardial injury via increasing superoxide dismutase activity and promoting myocardial active oxygen clearance. Besides, 3-NNMS can improve heart rate recovery and keep mitochondrial membrane integrity during the reperfusion period.

Key words: 3-nitro-N-methyl-salicylamide, myocardial ischemia/reperfusion injury, mitochondrial inhibitor, heart preservation solution, isolated heart preservation, reactive oxygen

CLC Number:

Wang Shuo, Liu Wenying, Lü Chaofan, Li Jiacong, Geng Yi, Zhao Yungang. Cardioprotective effect of 3-nitro-N-methyl salicylamide on the isolated rat heart under cold ischemia preservation[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(8): 1194-1201.

Figures/Tables 8

References

[1] SOARES ROS, LOSADA DM, JORDANI MC, et al. Ischemia/Reperfusion Injury Revisited: An Overview of the Latest Pharmacological Strategies. Int J Mol Sci. 2019;20(20):5034.
[2] CHOUCHANI ET, PELL VR, JAMES AM, et al. A Unifying Mechanism for Mitochondrial Superoxide Production during Ischemia-Reperfusion Injury. Cell Metab. 2016;23(2):254-263.
[3] MARIN W, MARIN D, AO X, et al. Mitochondria as a therapeutic target for cardiac ischemia‑reperfusion injury (Review). Int J Mol Med. 2021; 47(2):485-499.
[4] SUN MS, JIN H, SUN X, et al. Free Radical Damage in Ischemia-Reperfusion Injury: An Obstacle in Acute Ischemic Stroke after Revascularization Therapy. Oxid Med Cell Longev. 2018;2018:3804979.
[5] GONZÁLEZ-MONTERO J, BRITO R, GAJARDO AI, et al. Myocardial reperfusion injury and oxidative stress: Therapeutic opportunities. World J Cardiol. 2018;10(9):74-86.
[6] ELTZSCHIG HK, ECKLE T. Ischemia and reperfusion--from mechanism to translation. Nat Med. 2011;17(11):1391-1401.
[7] BERNARDI P, RASOLA A, FORTE M, et al. The Mitochondrial Permeability Transition Pore: Channel Formation by F-ATP Synthase, Integration in Signal Transduction, and Role in Pathophysiology. Physiol Rev. 2015; 95(4):1111-1155.
[8] DARE AJ, LOGAN A, PRIME TA, et al. The mitochondria-targeted anti-oxidant MitoQ decreases ischemia-reperfusion injury in a murine syngeneic heart transplant model. J Heart Lung Transplant. 2015; 34(11):1471-1480.
[9] BASILI S, TANZILLI G, MANGIERI E, et al. Intravenous ascorbic acid infusion improves myocardial perfusion grade during elective percutaneous coronary intervention: relationship with oxidative stress markers. JACC Cardiovasc Interv. 2010;3(2):221-229.
[10] PANISELLO-ROSELLÓ A, LOPEZ A, FOLCH-PUY E, et al. Role of aldehyde dehydrogenase 2 in ischemia reperfusion injury: An update. World J Gastroenterol. 2018;24(27):2984-2994.
[11] SOLIMAN S, ISHRAT T, FOUDA AY, et al. Sequential Therapy with Minocycline and Candesartan Improves Long-Term Recovery After Experimental Stroke. Transl Stroke Res. 2015;6(4):309-322.
[12] YU L, GONG B, DUAN W, et al. Melatonin ameliorates myocardial ischemia/reperfusion injury in type 1 diabetic rats by preserving mitochondrial function: role of AMPK-PGC-1α-SIRT3 signaling. Sci Rep. 2017;7:41337.
[13] CHEN C, LU W, WU G, et al. Cardioprotective effects of combined therapy with diltiazem and superoxide dismutase on myocardial ischemia-reperfusion injury in rats. Life Sci. 2017;183:50-59.
[14] Kalimeris K, Briassoulis P, Ntzouvani A, et al. N-acetylcysteine ameliorates liver injury in a rat model of intestinal ischemia reperfusion. J Surg Res. 2016;206(2):263-272.
[15] TSUJITA K, SHIMOMURA H, KAIKITA K, et al. Long-term efficacy of edaravone in patients with acute myocardial infarction. Circ J. 2006; 70(7):832-837.
[16] 徐建兴,钟永利,肖燕,等. 3-硝基-N-甲基水杨酰胺对琥珀酸-细胞色素C还原酶的抑制作用[J]. 生物化学杂志,1987,3(2):139-145.
[17] 沈明,徐建兴,陈清棠. 3-硝基-N-甲基-水杨酰胺对大鼠局灶脑缺血再灌注损伤的保护作用[J]. 中风与神经疾病,2002,19(1):17-19.
[18] WÖLKART G, SCHRAMMEL A, KOYANI CN, et al. Cardioprotective effects of 5-hydroxymethylfurfural mediated by inhibition of L-type Ca2+ currents. Br J Pharmacol. 2017;174(20):3640-3653.
[19] KWAN JC, GAO L, MACDONALD PS, et al. Cardio-protective signalling by glyceryl trinitrate and cariporide in a model of donor heart preservation. Heart Lung Circ. 2015;24(3):306-318.
[20] CHAMBERS DC, YUSEN RD, CHERIKH WS, et al. The Registry of the International Society for Heart and Lung Transplantation: Thirty-fourth Adult Lung And Heart-Lung Transplantation Report-2017;Focus Theme: Allograft ischemic time. J Heart Lung Transplant. 2017;36(10): 1047-1059.
[21] CADENAS S. ROS and redox signaling in myocardial ischemia-reperfusion injury and cardioprotection. Free Radic Biol Med. 2018; 117:76-89.
[22] VENKAT P, SHEN Y, CHOPP M, et al. Cell-based and pharmacological neurorestorative therapies for ischemic stroke. Neuropharmacology. 2018;134(Pt B):310-322.
[23] ZHOU H, MA Q, ZHU P, et al. Protective role of melatonin in cardiac ischemia-reperfusion injury: From pathogenesis to targeted therapy. J Pineal Res. 2018;64(3):10.1111/jpi.12471.
[24] ZHOU H, ZHANG Y, HU S, et al. Melatonin protects cardiac microvasculature against ischemia/reperfusion injury via suppression of mitochondrial fission-VDAC1-HK2mPTP-mitophagy axis. J Pineal Res. 2017;63(1): e12413.
[25] KALIMERIS K, BRIASSOULIS P, NTZOUVANI A, et al. N-acetylcysteine ameliorates liver injury in a rat model of intestinal ischemia reperfusion. J Surg Res. 2016;206(2):263-272.
[26] VALLS-LACALLE L, BARBA I, MIRÓ-CASAS E, et al. Succinate dehydrogenase inhibition with malonate during reperfusion reduces infarct size by preventing mitochondrial permeability transition. Cardiovasc Res. 2016;109(3):374-384.
[27] GUO L, SHESTOV AA, WORTH AJ, et al. Inhibition of Mitochondrial Complex II by the Anticancer Agent Lonidamine. J Biol Chem. 2016; 291(1):42-57.
[28] VANDEN HOEK TL, BECKER LB, SHAO Z, et al. Reactive oxygen species released from mitochondria during brief hypoxia induce preconditioning in cardiomyocytes. J Biol Chem. 1998;273(29): 18092-18098.
[29] SUZUKI C, HATAYAMA N, OGAWA T, et al. Cardioprotection via metabolism for rat heart preservation using the high-pressure gaseous mixture of carbon monoxide and oxygen. Int J Mol Sci. 2020; 21(22):8858.
[30] ELGEBALY SA, POSTON R, TODD R, et al. Cyclocreatine protects against ischemic injury and enhances cardiac recovery during early reperfusion. Expert Rev Cardiovasc Ther. 2019;17(9):683-697.
[31] YANG C, XU H, CAI L, et al. Donor pretreatment with adenosine monophosphate-activated protein kinase activator protects cardiac grafts from cold ischaemia/reperfusion injury. Eur J Cardiothorac Surg. 2016;49(5):1354-1360.
[32] WU MY, YIANG GT, LIAO WT, et al. Current mechanistic concepts in ischemia and reperfusion injury. Cell Physiol Biochem. 2018;46(4):1650-1667.
[33] ZOROVA LD, POPKOV VA, PLOTNIKOV EY, et al. Mitochondrial membrane potential. Anal Biochem. 2018; 552:50-59.
[34] ZHOU HY, ZHANG LN, ZHENG MZ, et al. Improved myocardial function with supplement of levosimendan to Celsior solution. J Cardiovasc Pharmacol. 2014;64(3):256-265.