Moderate cold acclimation (MCA) is a non-invasive intervention mitigating effects of various pathological conditions including myocardial infarction. We aim to determine the shortest cardioprotective regimen of MCA and the response of β1/2/3-adrenoceptors (β-AR), its downstream signaling, and inflammatory status, which play a role in cell-survival during myocardial infarction. Adult male Wistar rats were acclimated (9 °C, 1-3-10 days). Infarct size, echocardiography, western blotting, ELISA, mitochondrial respirometry, receptor binding assay, and quantitative immunofluorescence microscopy were carried out on left ventricular myocardium and brown adipose tissue (BAT). MultiPlex analysis of cytokines and chemokines in serum was accomplished. We found that short-term MCA reduced myocardial infarction, improved resistance of mitochondria to Ca2+-overload, and downregulated β1-ARs. The β2-ARs/protein kinase B/Akt were attenuated while β3-ARs translocated on the T-tubular system suggesting its activation. Protein kinase G (PKG) translocated to sarcoplasmic reticulum and phosphorylation of AMPKThr172 increased after 10 days. Principal component analysis revealed a significant shift in cytokine/chemokine serum levels on day 10 of acclimation, which corresponds to maturation of BAT. In conclusion, short-term MCA increases heart resilience to ischemia without any negative side effects such as hypertension or hypertrophy. Cold-elicited cardioprotection is accompanied by β1/2-AR desensitization, activation of the β3-AR/PKG/AMPK pathways, and an immunomodulatory effect.

1st Faculty of Medicine Center for Advanced Preclinical Imaging Charles University Prague Czech Republic

Centre of Experimental Medicine Institute for Heart Research Slovak Academy of Sciences Bratislava Slovak Republic

Faculty of Science Department of Physiology Charles University Vinicna 7 128 00 Prague 2 Czech Republic

Institute of Animal Physiology and Genetics Czech Academy of Sciences Libechov Czech Republic

Institute of Biotechnology Czech Academy of Sciences Prague West Czech Republic

Institute of Physiology Czech Academy of Sciences Prague Czech Republic

School of Pharmacy and Medical Science Griffith University Southport QLD Australia

Zobrazit více v PubMed

Heusch G. Critical issues for the translation of cardioprotection. Circ. Res. 2017;120:1477–1486. PubMed

Paradies, V., Chan, M. H. H. & Hausenloy, D. J. Strategies for reducing myocardial infarct size following STEMI. Primary Angioplasty 307–322. https://pubmed.ncbi.nlm.nih.gov/31314426/ (2018). PubMed

Hanssen MJW, et al. Short-term cold acclimation improves insulin sensitivity in patients with type 2 diabetes mellitus. Nat. Med. 2015;21:863–865. PubMed

Hanssen MJW, et al. Short-term cold acclimation recruits brown adipose tissue in obese humans. Diabetes. 2016;65:1179–1189. PubMed

KralovaLesna I, Rychlikova J, Vavrova L, Vybiral S. Could human cold adaptation decrease the risk of cardiovascular disease? J. Therm. Biol. 2015;52:192–198. PubMed

Sun Z, Cade R. Cold-induced hypertension and diuresis. J. Therm. Biol. 2000;25:105–109.

ClinicalTrials.gov. Chronic Cold Exposure and Energy Metabolism in Humans. https://beta.clinicaltrials.gov/study/NCT01730105 (U. S. National Library of Medicine, 2022).

ClinicalTrials.gov. The Effect of Cold Exposure on Energy Expenditure. https://beta.clinicaltrials.gov/study/NCT05107570 (U. S. National Library of Medicine, 2022).

ClinicalTrials.gov. Cold Acclimation as a Modulator of Brown Adipose Tissue Function in Adults with Obesity (MOTORBAT). https://beta.clinicaltrials.gov/study/NCT05468151 (U. S. National Library of Medicine, 2022).

Tibenska V, et al. The cardioprotective effect persisting during recovery from cold acclimation is mediated by the b2-adrenoceptor pathway and Akt activation. J. Appl. Physiol. 2021;130:746–755. PubMed

Tibenska V, et al. Gradual cold acclimation induces cardioprotection without affecting β-adrenergic receptor-mediated adenylyl cyclase signaling. J. Appl. Physiol. 2020;128:1023–1032. PubMed

Lefkowitz RJ, Rockman HA, Koch WJ. Catecholamines, cardiac β-adrenergic receptors, and heart failure. Circulation. 2000;101:1634–1637. PubMed

Scanzano A, Cosentino M. Adrenergic regulation of innate immunity: A review. Front. Pharmacol. 2015;6:1–18. PubMed PMC

Grisanti LA, et al. β2-Adrenergic receptor-dependent chemokine receptor 2 expression regulates leukocyte recruitment to the heart following acute injury. Proc. Natl. Acad. Sci. USA. 2016;113:15126–15131. PubMed PMC

Grisanti LA, et al. Leukocyte-expressed β2-adrenergic receptors are essential for survival after acute myocardial injury. Circulation. 2016;134:153–167. PubMed PMC

Brodde OE, Michel MC, Brodde OE, Michel MC. Adrenergic and muscarinic receptors in the human heart. Pharmacol. Rev. 1999;51:651–689. PubMed

Barki-Harrington L, Perrino C, Rockman HA. Network integration of the adrenergic system in cardiac hypertrophy. Cardiovasc. Res. 2004;63:391–402. PubMed

Lohse MJ, Engelhardt S, Eschenhagen T. What is the role of β-adrenergic signaling in heart failure? Circ. Res. 2003;93:896–906. PubMed

Balligand JL. Cardiac salvage by tweaking with beta-3-adrenergic receptors. Cardiovasc. Res. 2016;111:128–133. PubMed

Micova P, et al. Chronic intermittent hypoxia affects the cytosolic phospholipase A2α/cyclooxygenase 2 pathway via β2-adrenoceptor-mediated ERK/p38 stimulation. Mol. Cell. Biochem. 2016;423:151–163. PubMed

Barr LA, et al. Exercise training provides cardioprotection by activating and coupling endothelial nitric oxide synthase via a β3-adrenergic receptor-AMP-activated protein kinase signaling pathway. Med. Gas Res. 2017;7:1–8. PubMed PMC

Belge C, et al. Enhanced expression of β3-adrenoceptors in cardiac myocytes attenuates neurohormone-induced hypertrophic remodeling through nitric oxide synthase. Circulation. 2014;129:451–462. PubMed

Dubois-Deruy E, et al. Beta 3 adrenoreceptors protect from hypertrophic remodelling through AMP-activated protein kinase and autophagy. ESC Hear. Fail. 2020;7:920–932. PubMed PMC

Sentis SC, Oelkrug R, Mittag J. Thyroid hormones in the regulation of brown adipose tissue thermogenesis. Endocr. Connect. 2021;10:R106–R115. PubMed PMC

Nedergaard, J., Wang, Y. & Cannon, B. Cell proliferation and apoptosis inhibition: Essential processes for recruitment of the full thermogenic capacity of brown adipose tissue. Biochim. Biophys. Acta-Mol. Cell Biol. Lipids1864, 51–58 (2019). PubMed

Hanssen MJW, et al. Serum FGF21 levels are associated with brown adipose tissue activity in humans. Sci. Rep. 2015;5:1–8. PubMed PMC

Villarroya J, et al. New insights into the secretory functions of brown adipose tissue. J. Endocrinol. 2019;243:R19–R27. PubMed

Lømo T, Eken T, Bekkestad Rein E, Njå A. Body temperature control in rats by muscle tone during rest or sleep. Acta Physiol. 2020;228:1–26. PubMed

Shechtman O, Fregly MJ, Papanek PE. Factors affecting cold-induced hypertension in rats. Proc. Soc. Exp. Biol. Med. 1990;195:364–368. PubMed

Trappanese DM, et al. Chronic β1-adrenergic blockade enhances myocardial β3-adrenergic coupling with nitric oxide-cGMP signaling in a canine model of chronic volume overload: New insight into mechanisms of cardiac benefit with selective β1-blocker therapy. Basic Res. Cardiol. 2015;110:1–26. PubMed PMC

Schobesberger S, et al. β-Adrenoceptor redistribution impairs NO/cGMP/PDE2 signalling in 4 failing cardiomyocytes. Elife. 2020;9:1–15. PubMed PMC

Gauthier C, et al. The negative inotropic effect of β3-adrenoceptor stimulation is mediated by activation of a nitric oxide synthase pathway in human ventricle. J. Clin. Invest. 1998;102:1377–1384. PubMed PMC

Mongillo M, et al. Compartmentalized phosphodiesterase-2 activity blunts β-adrenergic cardiac inotropy via an NO/cGMP-dependent pathway. Circ. Res. 2006;98:226–234. PubMed

Farah C, Michel LYM, Balligand JL. Nitric oxide signalling in cardiovascular health and disease. Nat. Rev. Cardiol. 2018;15:292–316. PubMed

Cheng HJ, et al. Upregulation of functional β3-adrenergic receptor in the failing canine myocardium. Circ. Res. 2001;89:599–606. PubMed

Jin CZ, et al. Neuronal nitric oxide synthase is up-regulated by angiotensin II and attenuates NADPH oxidase activity and facilitates relaxation in murine left ventricular myocytes. J. Mol. Cell. Cardiol. 2012;52:1274–1281. PubMed

Calmettes G, et al. Hexokinases and cardioprotection. J. Mol. Cell. Cardiol. 2015;78:107–115. PubMed PMC

Halestrap AP, Pereira GC, Pasdois P. The role of hexokinase in cardioprotection—Mechanism and potential for translation. Br. J. Pharmacol. 2015;172:2085–2100. PubMed PMC

Waskova-Arnostova P, et al. Cardioprotective adaptation of rats to intermittent hypobaric hypoxia is accompanied by the increased association of hexokinase with mitochondria. J. Appl. Physiol. 2015;119:1487–1493. PubMed

Waskova-Arnostova P, et al. Chronic hypoxia enhances expression and activity of mitochondrial creatine kinase and hexokinase in the rat ventricular myocardium. Cell. Physiol. Biochem. 2014;33:310–320. PubMed

Costa ADT, Garlid KD. Intramitochondrial signaling: Interactions among mitoKATP, PKCε, ROS, and MPT. Am. J. Physiol.-Heart Circ. Physiol. 2008;295:H874–H882. PubMed PMC

Li X, et al. AMPK: A therapeutic target of heart failure—Not only metabolism regulation. Biosci. Rep. 2019;39:1–13. PubMed PMC

Marino A, et al. AMP-activated protein kinase: A remarkable contributor to preserve a healthy heart against ROS injury. Free Radic. Biol. Med. 2021;166:238–254. PubMed

Salminen A, Kaarniranta K. AMP-activated protein kinase (AMPK) controls the aging process via an integrated signaling network. Ageing Res. Rev. 2012;11:230–241. PubMed

Calvert JW, et al. Exercise protects against myocardial ischemia-reperfusion injury via stimulation of β3-adrenergic receptors and increased nitric oxide signaling: Role of nitrite and nitrosothiols. Circ. Res. 2011;108:1448–1458. PubMed PMC

Hermida N, et al. Cardiac myocyte β3-adrenergic receptors prevent myocardial fibrosis by modulating oxidant stress-dependent paracrine signaling. Eur. Heart J. 2018;39:888–898. PubMed

Horman S, Beauloye C, Vanoverschelde JL, Bertrand L. AMP-activated protein kinase in the control of cardiac metabolism and remodeling. Curr. Heart Fail. Rep. 2012;9:164–173. PubMed

Liu SQ, et al. Endocrine protection of ischemic myocardium by FGF21 from the liver and adipose tissue. Sci. Rep. 2013;3:1–11. PubMed PMC

Planavila A, et al. Fibroblast growth factor 21 protects against cardiac hypertrophy in mice. Nat. Commun. 2013;4:1–12. PubMed

Mann DL. Innate immunity and the failing heart: The cytokine hypothesis revisited. Circ. Res. 2015;116:1254–1268. PubMed PMC

Robert M, Miossec P. Effects of interleukin 17 on the cardiovascular system. Autoimmun. Rev. 2017;16:984–991. PubMed

Mills KHG. IL-17 and IL-17-producing cells in protection versus pathology. Nat. Rev. Immunol. 2023;23:38–54. PubMed PMC

Kanda T, Takahashi T. Interleukin-6 and cardiovascular diseases. Jpn. Heart J. 2004;45:183–193. PubMed

Elyasi A, et al. The role of interferon-γ in cardiovascular disease: An update. Inflamm. Res. 2020;69:975–988. PubMed

Trinchieri G. Interleukin-12 and the regulation of innate resistance and adaptive immunity. Nat. Rev. Immunol. 2003;3:133–146. PubMed

Deshmane SL, Kremlev S, Amini S, Sawaya BE. Monocyte chemoattractant protein-1 (MCP-1): An overview. J. Interf. Cytokine Res. 2009;29:313–325. PubMed PMC

Bousquenaud M, et al. Monocyte chemotactic protein 3 is a homing factor for circulating angiogenic cells. Cardiovasc. Res. 2012;94:519–525. PubMed

Liu J, et al. Eosinophils improve cardiac function after myocardial infarction. Nat. Commun. 2020;11:1–15. PubMed PMC

Sharma HS, Das DK. Role of cytokines in myocardial ischemia and reperfusion. Mediators Inflamm. 1997;6:175–183. PubMed PMC

Henein MY, Vancheri S, Longo G, Vancheri F. The role of inflammation in cardiovascular disease. Int. J. Mol. Sci. 2022;23:1–23. PubMed PMC

Szodoray P, et al. Th1/Th2 imbalance, measured by circulating and intracytoplasmic inflammatory cytokines—Immunological alterations in acute coronary syndrome and stable coronary artery disease. Scand. J. Immunol. 2006;64:336–344. PubMed

Zwaag J, Naaktgeboren R, Van Herwaarden AE, Pickkers P, Kox M. The effects of cold exposure training and a breathing exercise on the inflammatory response in humans: A pilot study. Psychosom. Med. 2022;84:457–467. PubMed PMC

Janský L, et al. Immune system of cold-exposed and cold-adapted humans. Eur. J. Appl. Physiol. Occup. Physiol. 1996;72:445–450. PubMed

Kasparova D, et al. Cardioprotective and nonprotective regimens of chronic hypoxia diversely affect the myocardial antioxidant systems. Physiol. Genomics. 2015;47:612–620. PubMed

Parulek J, Srámek M, Cerveanský M, Novotová M, Zahradník I. A cell architecture modeling system based on quantitative ultrastructural characteristics. Methods Mol. Biol. 2009;500:289–312. PubMed

Neckář J, et al. Selective replacement of mitochondrial DNA increases the cardioprotective effect of chronic continuous hypoxia in spontaneously hypertensive rats. Clin. Sci. 2017;131:865–881. PubMed

Porter C, et al. Human and mouse brown adipose tissue mitochondria have comparable UCP1 function. Cell Metab. 2016;24:246–255. PubMed PMC

Hahnova K, et al. β-Adrenergic signaling in rat heart is similarly affected by continuous and intermittent normobaric hypoxia. Gen. Physiol. Biophys. 2016;35:165–173. PubMed

Klevstig M, et al. Transgenic rescue of defective Cd36 enhances myocardial adenylyl cyclase signaling in spontaneously hypertensive rats. Pflugers Arch. Eur. J. Physiol. 2013;465:1477–1486. PubMed

Ihnatovych I, et al. Maturation of rat brain is accompanied by differential expression of the long and short splice variants of G(s)alpha protein: identification of cytosolic forms of G(s)alpha. J. Neurochem. 2001;79:88–97. PubMed

Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods. 2012;9:671–675. PubMed PMC

Kohutova J, et al. Anti-arrhythmic cardiac phenotype elicited by chronic intermittent hypoxia is associated with alterations in connexin-43 expression, phosphorylation, and distribution. Front. Endocrinol. (Lausanne) 2019;9:1–10. PubMed PMC

Manders EMM, Verbeek FJ, Aten JA. Measurement of co-localization of objects in dual-colour confocal images. J. Microsc. 1993;169:375–382. PubMed

Schindelin J, et al. Fiji: An open-source platform for biological-image analysis. Nat. Methods. 2012;9:676–682. PubMed PMC

R Core Team. R: A Language and Environment for Statistical Computing. https://www.r-project.org/ (R Foundation for Statistical Computing, 2021).

Sanz H, et al. drLumi: An open-source package to manage data, calibrate, and conduct quality control of multiplex bead-based immunoassays data analysis. PLoS One. 2017;12:1–18. PubMed PMC

KupcovaSkalnikova H, VodickovaKepkova K, Vodicka P. Luminex xMAP assay to quantify cytokines in cancer patient serum. Methods Mol. Biol. 2020;2108:65–88. PubMed

Gu Z, Eils R, Schlesner M. Complex heatmaps reveal patterns and correlations in multidimensional genomic data. Bioinformatics. 2016;32:2847–2849. PubMed

Vu, V. Q. GitHub—vqv/ggbiplot: A Biplot Based on ggplot2. https://github.com/vqv/ggbiplot (2011).

Acclimation of Hairless Spontaneously Hypertensive Rat to Ambient Temperature Attenuates Hypertension-Induced Pro-Arrhythmic Downregulation of Cx43 in the Left Heart Ventricle of Males