Exposure to sublethal heat stress activates a complex cascade of signaling events, such as activators (NO), signal molecules (PKCε), and mediators (HSP70 and COX-2), leading to implementation of heat preconditioning, an adaptive mechanism which makes the organism more tolerant to additional stress. We investigated the time frame in which these chemical signals are triggered after heat stress (41 ± 0.5°С/45 min), single or repeated (24 or 72 h after the first one) in heart tissue of male Wistar rats. The animals were allowed to recover 24, 48 or 72 h at room temperature. Single heat stress caused a significant increase of the concentration of HSP70, NO, and PKC level and decrease of COX-2 level 24 h after the heat stress, which in the next course of recovery gradually normalized. The second heat stress, 24 h after the first one, caused a significant reduction of the HSP70 levels, concentration of NO and PKCɛ, and significant increase of COX-2 concentration. The second exposure, 72 h after the first heat stress, caused more expressive changes of HSP70 and NO in the 24 h-recovery groups. The level of PKCɛ was not significantly changed, but there was significantly increased COX-2 concentration during recovery. Serum activity of AST, ALT, and CK was reduced after single exposure and increased after repeated exposure to heat stress, in both time intervals. In conclusion, a longer period of recovery (72 h) between two consecutive sessions of heat stress is necessary to achieve more expressive changes in mediators (HSP70) and triggers (NO) of heat preconditioning.
Single and repeated heat stress Activators Mediators Triggers Rat’s heart
This is a preview of subscription content, log in to check access.
We thank prof. Michal Horowitz (Laboratory of Environmental Physiology, The Hebrew University, Hadassah Medical Center, Jerusalem, Israel) for advice and critical comments during writing the paper.
The research was performed at the Department of Experimental Physiology and Biochemistry, Institute of Biology, Faculty of Natural Sciences and Mathematics, University “Ss Cyril and Methodius”, Skopje, R. Macedonia.
This research did not receive any specific grant from funding agencies in the public, commercial or non-profit sectors.
Compliance with ethical standards
Conflict of interest
We confirm that there is no conflict of interest between authors.
Arakawa H, Kodama H, Matsuoka N, Yamaguchi I (1997) Stress increases plasma enzyme activity in rats: differential effects of adrenergic and cholinergic blockades. J Pharmacol Exp Ther 280(3):1296–1303PubMedGoogle Scholar
Arnaud C, Laubriet A, Joyeux M, Godin-Ribuot D, Rochette L, Demenge P, Ribuot C (2001) Role of nitric oxide synthases in the infarct size-reducing effect conferred by heat stress in isolated rat hearts. Br J Pharmacol 132(8):1845–1851CrossRefPubMedPubMedCentralGoogle Scholar
Arnaud C, Joyeux M, Garrel C, Godin-Ribuot D, Demenge P, Ribuot C (2002) Free-radical production triggered by hyperthermia contributes to heat stress-induced cardioprotection in isolated rat hearts. Br J Pharmacol 135(7):1776–1782Google Scholar
Arnaud C, Godin-Ribuot D, Bottari S, Peinnequin A, Joyeux M, Demenge P, Ribuot C (2003a) iNOS is a mediator of the heat stress-induced preconditioning against myocardial infarction in vivo in the rat. Cardiovasc Res 58(1), pp.118–125Google Scholar
Arnaud C, Joyeux-Faure M, Godin-Ribuot D, Ribuot C (2003b) COX-2: an in vivo evidence of its participation in heat stress-induced myocardial preconditioning. Cardiovasc Res 58(3):582–588Google Scholar
Bolli R (1996) The early and late phases of preconditioning against myocardial stunning and the essential role of oxyradicals in the late phase: an overview. In: New paradigms of coronary artery disease. Steinkopff, Heidelberg, pp 175–181Google Scholar
Bolli R, Dawn B, Tang XL, Qiu Y, Ping P, Xuan YT, Jones WK, Takano H, Guo Y, Zhang J (1998) The nitric oxide hypothesis of late preconditioning. Basic Res Cardiol 93(5):325–338CrossRefPubMedPubMedCentralGoogle Scholar
Das M, Das DK (2008) Molecular mechanism of preconditioning. Int Union Biochem Mol Biol Life 60(4):199–203CrossRefGoogle Scholar
DeMarco VG, Scumpia PO, Bosanquet JP, Skimming JW (2004) α-lipoic acid inhibits endotoxin-stimulated expression of iNOS and nitric oxide independent of the heat shock response in RAW 264.7 cells. Free Radic Res 38(7):675–682CrossRefPubMedGoogle Scholar
Diao LW, Zhao LL, Qi F, Sun ZD, Zhang QH, Wu NS (2012) Heat shock protein 70 induced by heat stress protects heterotopically transplanted hearts in rats. Mol Med Rep 6(4):729–732CrossRefPubMedGoogle Scholar
Franesoni RP, Mager M (1978) Heat injured rats: pathochemical indices and survival time. J Appl Physiol 45:1–6CrossRefGoogle Scholar
Goto K, Kojima A, Kobayashi T, Uehara K, Morioka S, Naito T, Akema T, Sugiura T, Ohira Y, Yoshioka T (2005) Heat stress as a countermeasure for prevention of muscle atrophy in microgravity environment. Jpn J Aerosp Environ Med 42(2):51–59Google Scholar
Horowitz M (2007) Heat acclimation and cross-tolerance against novel stressors: genomic–physiological linkage. Prog Brain Res 162:373–392CrossRefPubMedGoogle Scholar
Horowitz M, Eli-Berchoer L, Wapinski I, Friedman N, Kodesh E (2004) Stress-related genomic responses during the course of heat acclimation and its association with ischemic-reperfusion cross-tolerance. J Appl Physiol 97(4):1496–1507CrossRefPubMedGoogle Scholar
Hoshida S, Yamashita N, Otsu K, Hori M (2002) Repeated physiologic stresses provide persistent cardioprotection against ischemia-reperfusion injury in rats. J Am Coll Cardiol 40(4):826–831CrossRefPubMedGoogle Scholar
Howard M, Roux J, Lee H, Miyazawa B, Lee JW, Gartland B, Howard AJ, Matthay MA, Carles M, Pittet JF (2010) Activation of the stress protein response inhibits the STAT1 signaling pathway and iNOS function in alveolar macrophages: role of Hsp90 and Hsp70. Thorax 65(4):346–353CrossRefPubMedPubMedCentralGoogle Scholar
Hsu SF, Chao CM, Huang WT, Lin MT, Cheng BC (2013) Attenuating heat-induced cellular autophagy, apoptosis and damage in H9c2 cardiomyocytes by pre-inducing HSP70 with heat shock preconditioning. Int J Hyperth 29(3):239–247CrossRefGoogle Scholar
Joyeux M, Godin-Ribuot D, Yellon DM, Demenge P, Ribuot C (1999) Heat stress response and myocardial protection. Fundam Clin Pharmacol 13(1):1–10CrossRefPubMedGoogle Scholar
Joyeux-Faure M, Arnaud C, Godin-Ribuot D, Ribuot C (2003) Heat stress preconditioning and delayed myocardial protection: what is new? Cardiovasc Res 60(3):469–477CrossRefPubMedGoogle Scholar
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275PubMedGoogle Scholar
Malyshev IY, Manukhina EB, Mikoyan VD, Kubrina LN, Vanin AF (1995) Nitric oxide is involved in heat-induced HSP70 accumulation. FEBS Lett 370(3):159–162CrossRefPubMedGoogle Scholar
Maslov LN, Khaliulin IG, Zhang I (2010) Role of heat shock proteins in the mechanism of cardioprotective effect of transient hyperthermia and delayed preconditioning. Patol Fiziol Eksp Ter 4:64–73Google Scholar
Miova B, Dinevska-Kjovkarovska S, Esplugues JV, Apostolova N (2015) Heat stress induces extended plateau of Hsp70 accumulation–a possible Cytoprotection mechanism in hepatic cells. J Cell Biochem 116(10):2365–2374CrossRefPubMedGoogle Scholar
Oka Y, Akagi Y, Kinugasa T, Ishibashi N, Iwakuma N, Shiratsuchi I, Shirouzu K (2013) Heat-shock pre-treatment reduces liver injury and aids liver recovery after partial hepatectomy in mice. Anticancer Res 33(7):2887–2894PubMedGoogle Scholar
Patel HH, Hsu A, Gross GJ (2001) Cardioprotection is strain dependent in rat in response to whole body hyperthermia. Am J Phys Heart Circ Phys 280(3):H1208–H1214Google Scholar
Ping P, Takano H, Zhang J, Tang XL, Qiu Y, Li RC, Banerjee S, Dawn B, Balafonova Z, Bolli R (1999) Isoform-selective activation of protein kinase C by nitric oxide in the heart of conscious rabbits a signaling mechanism for both nitric oxide–induced and ischemia-induced preconditioning. Circ Res 84(5):587–604CrossRefPubMedGoogle Scholar
Qian YZ, Shipley JB, Levasseur JE, Kukreja RC (1998) Dissociation of heat shock proteins expression with ischemic tolerance by whole body hyperthermia in rat heart. J Mol Cell Cardiol 30(6):1163–1172CrossRefPubMedGoogle Scholar
Shinmura K, Tang XL, Wang Y, Xuan YT, Liu SQ, Takano H, Bhatnagar A, Bolli R (2000) Cyclooxygenase-2 mediates the cardioprotective effects of the late phase of ischemic preconditioning in conscious rabbits. Proc Natl Acad Sci 97(18):10197–10202CrossRefPubMedGoogle Scholar
Somji S, Todd JH, Sens MA, Garrett SH, Sens DA (1999) Expression of the constitutive and inducible forms of heat shock protein 70 in human proximal tubule cells exposed to heat, sodium arsenite, and CdCl (2). Environ Health Perspect 107(11):887PubMedPubMedCentralGoogle Scholar
Velkovski M (2012) Effects of heat stress and nicotinamide on induction of HSP72 mRNA in heart of diabetic rats. Master thesis, University “Ss Cyril and Methodius”, SkopjeGoogle Scholar
Xi D, Tekin P, Bhargava RC, Kukreja L (2001) Whole body hyperthermia and preconditioning of the heart: basic concepts, complexity, and potential mechanisms. Int J Hyperth 17(5):439–455CrossRefGoogle Scholar
Yamashita N, Hoshida S, Taniguchi N, Kuzuya T, Hori M (1998) Whole-body hyperthermia provides biphasic cardioprotection against ischemia/reperfusion injury in the rat. Circulation 98(14):1414–1421CrossRefPubMedGoogle Scholar
Yang YL, Lin MT (1999) Heat shock protein expression protects against cerebral ischemia and monoamine overload in rat heatstroke. Am J Phys Heart Circ Phys 276(6):H1961–H1967Google Scholar
Zhang L, Liu Q, Yuan X, Wang T, Luo S, Lei H, Xia Y (2013) Requirement of heat shock protein 70 for inducible nitric oxide synthase induction. Cell Signal 25(5):1310–1317CrossRefPubMedGoogle Scholar