Isoflurane ameliorates oxygen-glucose deprivation-induced cardiomyocyte injury through SIRT6/DNMT1 pathway

The incidence of cardiovascular diseases is on the rise in the world, which poses a significant threat to human health. Myocardial ischemia can cause heart disease. Therefore, it is necessary to avoid myocardial hypoxia/reoxygenation (H/R) injury to attenuate the risk of heart disease. The present study focuses on the protective effect of isoflurane on H/R-induced cell injury through the Sirtuin 6 (SIRT6)/DNA (cytosine-5)-methyltransferase 1 (DNMT1) pathway. Quantitative reverse transcription PCR (RT‑qPCR) and Western blot analysis were used to measure protein levels and mRNA expression in H9c2 cells. Cell Counting Kit‑8 assays (CCK8 assay) was used to determine cell viability. The expression levels of pro-inflammatory molecule were assessed using commercial Enzyme-linked immunosorbent assay (ELISA) Kits. The ratio of cellular apoptosis was determined by flow cytometry. The contents of Lactate dehydrogenase (LDH), Cardiac Troponin I (cTnI), and Creatine Kinase MB (CK-MB) were detected using colorimetric assays. This study shows that Isoflurane reduces the expression of DNMT1 by activating SIRT6 in oxygen-glucose deprivation (OGD)-induced H/R injury. The damage of cardiomyocyte was decreased after Isoflurane treatment under OGD exposure condition. In addition, Isoflurane ameliorates OGD-induced inflammatory responses and cellular apoptosis in H9c2 cell via interaction with the SIRT6/DNMT1 pathway. Taken together, this study suggested the protective effect of Isoflurane on the process of OGD-induced damage and provided a new mechanism of action for Isoflurane in the treatment of H/R-induced cardiomyocyte injury.

Isoflurane; OGD; SIRT6; DNMT1; Hypoxia/reoxygenation

[1] Thomas H, Diamond J, Vieco A, Chaudhuri S, Shinnar E, Cromer S, et al. Global atlas of cardiovascular disease. Global Heart. 2018; 13: 143–163.

[2] Khan MA, Hashim MJ, Mustafa H, Baniyas MY, Al Suwaidi SKBM, AlKatheeri R, et al. Global epidemiology of ischemic heart disease: results from the global burden of disease study. Cureus. 2020; 12: e9349.

[3] Wei H, Liang G, Yang H. Isoflurane preconditioning inhibited isoflurane-induced neurotoxicity. Neuroscience Letters. 2007; 425: 59–62.

[4] Constantinides C, Mean R, Janssen BJ. Effects of isoflurane anes-thesia on the cardiovascular function of the C57BL/6 mouse. ILAR Journal/National Research Council, Institute of Laboratory Animal Resources. 2011; 52: e21–e31.

[5] Liu J, Yang S, Zhang X, Liu G, Yue X. Isoflurane reduces oxygen-glucose deprivation-induced oxidative, inflammatory, and apoptotic responses in H9c2 cardiomyocytes. American Journal of Translational Research. 2016; 8: 2597.

[6] Su Y, Chen G, Zhang F, Wang L, Feng Z, Gao X. Isoflurane alleviates myocardial injury induced by hypoxia/reoxygenation by regulating miR-18a-5p. Cardiovascular Toxicology. 2021; 21: 800–807.

[7] Trapp J. The role of NAD+ dependent histone deacetylases (sirtuins) in ageing. Current Drug Targets. 2006; 7: 1553–1560.

[8] Mendes KL, de Farias Lelis D, Santos SHS. Nuclear sirtuins and inflammatory signaling pathways. Cytokine & Growth Factor Reviews. 2017; 38: 98–105.

[9] Poulose N, Raju R. Sirtuin regulation in aging and injury. Biochimica Et Biophysica Acta (BBA)—Molecular Basis of Disease. 2015; 1852: 2442–2455.

[10] Tennen RI, Chua KF. Chromatin regulation and genome maintenance by mammalian SIRT6. Trends in Biochemical Sciences. 2011; 36: 39–46.

[11] Wang X, Wang X, Tong M, Gan L, Chen H, Wu S, et al. SIRT6 protects cardiomyocytes against ischemia/reperfusion injury by augmenting FoxO3α-dependent antioxidant defense mechanisms. Basic Research in Cardiology. 2016; 111: 13.

[12] Chen G, Zhang F, Wang L, Feng Z. Isoflurane alleviates hypoxia/reoxygenation induced myocardial injury by reducing miR-744 mediated SIRT6. Toxicology Mechanisms and Methods. 2022; 32: 235–242.

[13] Deng Z, Yao J, Xiao N, Han Y, Wu X, Ci C, et al. DNA methyltransferase 1 (DNMT1) suppresses mitophagy and aggravates heart failure via the microRNA-152-3p/ETS1/RhoH axis. Laboratory Investigation. 2022; 102: 782–793.

[14] Boovarahan SR, Kurian GA. Investigating the role of DNMT1 gene expression on myocardial ischemia reperfusion injury in rat and associated changes in mitochondria. Biochimica Et Biophysica Acta (BBA)—Bioenergetics. 2022; 1863: 148566.

[15] Jia B, Chen J, Wang Q, Sun X, Han J, Guastaldi F, et al. SIRT6 promotes osteogenic differentiation of adipose-derived mesenchymal stem cells through antagonizing DNMT1. Frontiers in Cell and Developmental Biology. 2021; 9: 648627.

[16] Bagheri F, Khori V, Alizadeh AM, Khalighfard S, Khodayari S, Khodayari H. Reactive oxygen species-mediated cardiac-reperfusion injury: mechanisms and therapies. Life Sciences. 2016; 165: 43–55.

[17] Zare MFR, Rakhshan K, Aboutaleb N, Nikbakht F, Naderi N, Bakhshesh M, et al. Apigenin attenuates doxorubicin induced cardiotoxicity via reducing oxidative stress and apoptosis in male rats. Life Sciences. 2019; 232: 116623.

[18] Qiu Z, Lei S, Zhao B, Wu Y, Su W, Liu M, et al. NLRP3 inflammasome activation-mediated pyroptosis aggravates myocardial ischemia/reperfusion injury in diabetic rats. Oxidative Medicine and Cellular Longevity. 2017; 2017: 1–17.

[19] Kanazawa M, Miura M, Toriyabe M, Koyama M, Hatakeyama M, Ishikawa M, et al. Microglia preconditioned by oxygen-glucose depri-vation promote functional recovery in ischemic rats. Scientific Reports. 2017; 7: 1–16.

[20] Kurian GA, Rajagopal R, Vedantham S, Rajesh M. The role of oxidative stress in myocardial ischemia and reperfusion injury and remodeling: revisited. Oxidative Medicine and Cellular Longevity. 2016; 2016: 1–14.

[21] Li C, Miao X, Li F, Adhikari BK, Liu Y, Sun J, et al. Curcuminoids: implication for inflammation and oxidative stress in cardiovascular diseases. Phytotherapy Research. 2019; 33: 1302–1317.

[22] Relja B, Land WG. Damage-associated molecular patterns in trauma. European Journal of Trauma and Emergency Surgery. 2020; 46: 751–775.

[23] Anselmi A. Myocardial ischemia, stunning, inflammation, and apoptosis during cardiac surgery: a review of evidence. European Journal of Cardio-Thoracic Surgery. 2004; 25: 304–311.

[24] Olson JM, Yan Y, Bai X, Ge Z, Liang M, Kriegel AJ, et al. Up-regulation of MicroRNA-21 mediates isoflurane-induced protection of cardiomyocytes. Anesthesiology. 2015; 122: 795–805.

[25] Sayed S, Idriss NK, Sayyed HG, Ashry AA, Rafatt DM, Mohamed AO, et al. Effects of propofol and isoflurane on haemodynamics and the inflammatory response in cardiopulmonary bypass surgery. British Journal of Biomedical Science. 2015; 72: 93–101.

[26] Zhang S, Zhang Y. Isoflurane reduces endotoxin-induced oxidative, inflammatory, and apoptotic responses in H9c2 cardiomyocytes. European Review for Medical and Pharmacological Sciences. 2018; 22: 3976–3987.

[27] Wei H, Kang B, Wei W, Liang G, Meng QC, Li Y, et al. Isoflurane and sevoflurane affect cell survival and BCL-2/BAX ratio differently. Brain Research. 2005; 1037: 139–147.

[28] Jamnicki-Abegg M, Weihrauch D, Pagel P, Kersten J, Bosnjak Z, Warltier D, et al. Isoflurane inhibits cardiac myocyte apoptosis during oxidative and inflammatory stress by activating akt and enhancing Bcl-2 expression. Anesthesiology. 2005; 103: 1006–1014.

[29] Yang Y, Tian T, Wang Y, Li Z, Xing K, Tian G. SIRT6 protects vascular endothelial cells from angiotensin II-induced apoptosis and oxidative stress by promoting the activation of Nrf2/ARE signaling. European Journal of Pharmacology. 2019; 859: 172516.