Liver ischemia preconditions the heart against ischemia-reperfusion arrhythmias

Document Type: Original Article


1 Department of Pharmacology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

2 Department of Physiology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran


Objective(s):This study aimed to examine the hypothesis that an antiarrhythmic effect might be obtained by ischemic preconditioning of the liver, and also to characterize the potential underlying mechanisms.
Materials and Methods: Male Wistar rats were anesthetized by thiopental sodium (50 mg/kg, IP) followed by IV injection of heparin (250 IU). Remote ischemic preconditioning (RIPC) was induced by 3 cycles of 5 min liver ischemia followed by 5 min of reperfusion. The hearts were excised within 5 min after the final cycle of preconditioning and perfused using Langendorff’s system.            The isolated perfused hearts were subjected to 30 min global ischemia followed by 90 min reperfusion. The myocardial arrhythmias induced by ischemia- reperfusion (I/R) were determined in accordance with the guidelines of Lambeth Conventions. The potential role of KATP channels on RIPC was assessed by injection of glibenclamide (nonselective KATP blocker) or             5-hydroxydecanoate (mitochondrial KATP blocker) on rats 30 and 15 min before induction of RIPC in the liver, respectively.
Results: Hepatic remote preconditioning of the heart significantly (P<0.0001) prevented the incidence of myocardial arrhythmias induced by I/R in the perfused hearts (5.33±1.54 vs. 32.33±6.44,). However, the protective effects of remote preconditioning was significantly (P<0.01) abolished by the KATP blocker, glibenclamide (25.5±4.9 vs. 5.33±1.54,).
Conclusion: Hepatic RIPC may prevent the arrhythmias induced by I/R in the isolated perfused hearts via KATP channels.


1.Bril A. Cellular mechanisms of cardiac arrhythmias in the ischemic and reperfused heart. EXS 1996; 76:135-153.

2.Mccord JM. Oxygen-derived free radicals in postischemic tissue injury. N Engl J Med 1985; 312:159-163.

3.Kloner RA, Przyklenk K, Whittaker P. Deleterious effects of oxygen radicals in ischemia/reperfusion. Resolved and unresolved issues. Circulation 1989; 80:1115-1127.

4.Dhalla NS, Elmoselhi AB, Hata T, Makino N. Status of myocardial antioxidants in ischemia-reperfusion injury. Cardiovasc Res 2000; 47:446-456.

5.Yang CS, Tsai PJ, Chou ST, Niu YL, Lai JS, Kuo JS. The roles of reactive oxygen species and endogenous opioid peptides in ischemia-induced arrhythmia of isolated rat hearts. Free Radic Biol Med 1995; 18:593-598.

6.Ravingerova T, Slezak J, Tribulova J, Dzurba A, Uhrik B, Ziegelhoffer A. Reactive oxygen species contribute to high incidence of reperfusion-induced arrhythmias in isolated rat heart. Life Sci 1999; 65:1927-1930.

7.Downey J, Omar B, Ooiwa H, Mccord J. Superoxide dismutase therapy for myocardial ischemia. Free Radic Res Commun 1991; 12:703-720.

8.Tang LD, Tang ZM. Protective effects of SH-compounds on ischemia reperfusion induced arrhythmias in the isolated rat heart. Yao Xue Xue Bao 1991; 26:91-95.

9.Qiuy, Galinanes M, Ferrari R, Cargnoni A, Ezrin A, Hearse DJ. PEG-SOD improves post-ischemic functional recovery and antioxidant status in blood-perfused rabbit hearts. Am J Physiol 1992; 263: H1243-H1249.

10.Murry CE, Jennings RB, Reimer KA. Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium. Circulation 1986; 74: 1124-1136.

11.Li,Y, P.Whittaker, Kloner R. The transient nature of the effect of ischemic preconditioning on myocardial infarct size and ventricular arrhythmia. Am Heart J 1992; 123:346–353.

12.Liu Y, Downey JM. Ischemic preconditioning protects against infarction in rat heart. Am J Physiol 1992; 263: H1107–H1112.

13.Murry, CE, Richard VJ, Reimer KA, Jennings RB. Ischemic preconditioning slows energy metabolism and delays ultrastructural damage during a sustained ischemic episode. Circ Res 1990; 66:913–931.

14.Efstathios KI, Antigone L, Dimitrios TK. Ischemic preconditioning: Protection against myocardial necrosis and apoptosis. Vasc Health Risk Manag 2007; 3: 629–637.

15.Walker MJA, Curtis MJ, Hearse DJ, Campbell RWF, Janse MJ, Yellon DM, et al. The Lambeth Conventions: guidelines for the study of arrhythmias in ischaemia, infarction, and reperfusion. Cardiovasc Res 1988; 22: 441-455.

16.Takayuki M, Uno I, Munetaka F, Jun S, Hirofumi A, Kazu-ichi Y. Effects of ischemic and Sevoflurane-induced preconditioning on myocardial infarction and arrhythmias in rabbits in vivo. J Anesth Clin Res 2013; 4:361-366.

17.Matejikova  J, Kucharska J, Pinterova M , Pancza D, Ravingerova T. Protection against ischemia-induced ventricular arrhythmias and myocardial dysfunction conferred by preconditioning in the rat heart: involvement of mitochondrial KATPChannels. Physiol Res 2009; 58:9-19.

18.Evrengul H, Seleci D, Tanriverdi H, Kaftan A. The antiarrhythmic effect and clinical consequences of ischemic preconditioning. Coron Artery Dis 2006 May; 17:283-288.

19.Derek J, Hausenloy M, Derek MY. Remote ischaemic preconditioning: underlying mechanisms and clinical application. Cardiovascular Research 2008; 79:377–386.

20.Przyklenk K, Bauer B, Ovize M, Kloner R, Whittaker P. Regional ischemic preconditioning protects remote virgin myocardium from subsequent sustained coronary occlusion. Circulation 1993; 87: 893–899.

21.Jose FC, Ricardo F-C, Adelino FLM. Myocardial remote ischemic preconditioning: From pathophysiology to clinical application. Rev Port Cardiol 2013; 32:893-904.

22.Das DK, Maulik A. Preconditioning potentiates redox signaling and converts death signal into survival signal. Arch Biochem Biophys 2003; 420:305-311.

23.Gross G, Fryer RM. Sarcolemmal versus mitochondrial ATP-sensitive K+ channels and myocardial preconditioning in dogs. Circ Res 1999; 9: 973-979.

24.Das B, Sarkar CH. Is the sarcolemmal or mitochondrial KATP channel activation important in the antiarrhythmic and cardioprotective effects during acute ischaemia/reperfusion in the intact anesthetized rabbit model. Life Sci 2005; 77: 1226-1248.

25.Vegh A, Parratt JR. The role of mito KATP             channels in antiarrhythmic effects of ischaemic preconditioning in dogs. Br J Pharmacol 2002; 137: 1107-1115.

26.Gross GJ, Auchampach JA. Blockade of ATP-sensitive potassium channel prevents myocardial preconditioning in dogs. Circ Res 1992; 70: 223-233.

27.Grover GJ, D’alonzo AJ, Sleph PG, Dzwonczyk S,            Hess T, Darbenzio RB. The cardioprotective and electrophysiological effects of cromakalim are attenuated by meclofenamate through a cyclooxygenase independent mechanism. J Pharmacol Exp 1994; 269: 536-540.

28.Tosaki A, Szerdahelyi P, Das DK. Reperfusion-induced arrhythmias and myocardial ion shifts: a pharmacologic interaction between pinacidil and cicletanine in isolated rat hearts. Basic Res Cardiol 1992; 87: 366-384.

29.Hamada K, Yamazaki J, Nagao T. Shortening of action potential duration is not prerequisite for cardiac protection by ischemic preconditioning or a KATP channel opener. J Mol Cell Cardiol 1998; 30: 1369-1379.

30.Ravingerova T, Lokebo JE, Sundset R, Ytrehus K. Preconditioning against contractile dysfunction in guinea pig papillary muscle depends on the opening of KATP-sensitive channels. Exp Clin Cardiol 1998; 3: 184-188.

31.Sato T, Sasaki N, Seharaseyon J, O’rourke B, Marban E. Selective pharmacological agents implicate mitochondrial but not sarcolemmal KATP channels in ischemic cardioprotection. Circulation 2000; 101: 2418-2423.

32.Munch-Ellingsen J, Lokebo JE, Bugge E, Jonassen AK, Ravingerova T, Ytrehus K: 5-HD abolishes ischemic preconditioning independently of monophasic action potential duration in the heart. Basic Res Cardiol 2000; 95: 228-234.

33.Garlid KD, Dos Santos P, Xie Z-J, Costa ADT, Paucek P. Mitochondrial potassium transport: the role of the mitochondrial ATP-sensitive K channels in cardiac function and cardioprotection. Biochim Biophys Acta 2003; 1606: 1-21.

34.Yue Y, Qin Q, Cohen MV, Downey JM, Critz SD. The relative order of mito KATP channels, free radicals and p38 MAPK in preconditioning’s protective pathway in rat heart. Cardiovasc Res 2002; 55: 681-689.

35.Hausenloy DJ, Yellon DM. Preconditioning and postconditioning: united at reperfusion. Pharmacol Ther 2007; 116: 173–191.

36.Steen BK, Ole H, Rajesh KK, Jens E N-K, Michael RS, Andrew N. Remote preconditioning reduces ischemic injury in the explanted heart by a KATP channel-dependent mechanism. Am J Physiol Heart Circ Physiol 2005; 288: H1252–H1256.

37.Harlokesh N Y, Manjeet S, P.L. Sharma, Dhiraj M, Tapan B, Atinder P K. Possible Role of Cyclooxy-genase-2 in Remote Aortic Preconditioning Induced Cardioprotection in Rat Heart. Pharmacologia 2012; 3: 1-8.

38.Lloris-Carsi JM, Cejalvo D, Toledo-Pereyra LH, Calvo MA, Suzuki S. Preconditioning: Effect upon lesion modulation in warm liver ischemia. Transplant Proc 1993; 25: 3303–3304.

39.Najafi M, Garjani A. The effect of L-carnitine on arrhythmias in the ischemic rat heart. Iran J Basic Med Sci 2005; 8: 38-44.

40.Najafi M, Shaseb E, Ghaffary Sa, Fakhrju A, Oskouei Eteraf T. Effects of chronic oral administration of natural honey on ischemia-reperfusion-induced arrhythmias in isolated rat heart. Iran J Basic Med Sci 2011; 14:75-81.

41.Shahid M, Tauseef M, Sharma K K, Fahim M. Brief femoral artery ischemia provides protection agains myocardial ischaemia–reperfusion injury in rats: the possible mechanisms. Exp Physiol 2008; 93: 954–968.

42.Przyklenk K, Whittaker P. Remote ischemic preconditioning current knowledge, unresolved questions, and future priorities. J Cardiovasc Pharmacol Ther 2011; 16: 255-259.

43.Clanahan MC, Nao B, Wolke L, Martin BJ, Mezt TE. Brief renal occlusion and reperfusion reduces myocardial infarct size in rabbits. FASEB J 1993; 7: A18.

44.Gho BC, Schoemaker RG, van den Doel MA, Duncker DJ, Verdouw PD. Myocardial protection by brief ischemia in noncardiac tissue. Circulation 1996; 94: 2193-2200.

45.Tang ZL, Dai W, Li YJ, Deng HW. Involvement of capsaicin-sensitive sensory nerves in early and delayed cardioprotection induced by a brief ischaemia of the small intestine. Naunyn Schmiedebergs Arch Pharmacol 1999; 359: 243-247.

46.Tatyana O, Michael A, Rodica K, Natalie A, Babeth R. Limb ischemia preconditions the heart against reperfusion tachyarrhythmia. Am J Physiol Heart Circ Physiol 1997; 273: H1707-H1712.

47.Heidi LL, Stephen ED. Partial Hind limb Occlusion Reduced the Susceptibility to Sustained Ventricular Tachycardia in Conscious Rats. J Cardiovasc Pharmacol Ther 2009; 14(3): 199–206.

48.Curtis MJ. Characterisation, utilisation and clinical relevance of isolated perfused heart models of ischemia-induced ventricular fibrillation. Cardiovasc Res 1998; 9: 194-215.

49.Bernier M, Ccurtis MJ, Hearse DJ. Ischemia-induced and reperfusion-induced arrhythmias: importance of heart rate. Am J Physiol 1989; 256: H21-H31.

50.Ravigerova T, Tibulova N, Slezak J, Curtis MJ. Brief intermediate and prolonged ischemia in the isolated crystalloid perfused rat heart: relationship between susceptibility to arrhythmias and degree of ultrastructural injury. J Mol Cell Cardiol 1995; 27: 1937-1951.

51.Botsford MW, Lukas A. Ishemic preconditioning and arrhythmogenesis in the rabbit heart: effects on epicardium vs. endocardium. J Mol Cell Cardiol 1998; 30: 1723-1735.

52.Cohen MV, Yang X-M, Liu GS, Heusch G, Downey JM. Acetylcholine, bradykinin, opioids, and henylephrine, but not adenosine, trigger preconditioning by generating free radicals and openingmitochondrial KATP channels. Circ Res 2001; 89: 273-278. 

53.Zhang DX, Chen YF, Campbell WB, Zou AP, Gross GJ, Li PL. Characteristics and superoxide-induced activation of reconstituted myocardial ATP-sensitive potassium channels. Circ Res 2001; 89: 1177-1183.

54.Kuazaki T, Masahiro A, Hitoshi I, Tomoaki T, Mikio Yanase, Yukiko I, et al. Ischemic Precondi-tioning Protects Hepatocytes Via Reactive Oxygen Species Derived From Kupffer Cells in Rats. Gastroenterology 2004; 127: 1488–1496.

55.Morihira M, Hasebe N, Baljinnyam E, Sumitomo K, Matsusaka T, Izawa K, et al. Ischemic preconditioning enhances scavenging activity of reactive oxygen species and diminishes transmural difference of infarct size. Am J Physiol 2006; 290: H577-H583.

56.Maczewski M, Duda M, Pawlak W, Reresewiks A. Endothelial protection from reperfusion injury by ischemic preconditioning and diazoxide involves a SOD-like anti-O2-mechanism. J Physiol Pharmacol 2004; 55: 537-550.

57.Glantz L, Avramovich A, Trembovler V, Gurvits V, Kohen R, Eidelman LA, ShohaMI E. Ischemic preconditioning increases antioxidants in the brain and peripheral organ after cerebral ischemia. Exp Neurol 2005; 192: 117-124.

58.Honda HM, Korge P, Weiss JN. Mitochondria and ischemia/reperfusion injury. Ann NY Acad Sci 2005; 1047: 248-258.

59.Glanemann M, Vollmar B, Nussler AK, Schaefer T, Neuhaus P, Menger MD. Ischemic preconditioning protects from hepatic ischemia/reperfusion-injury by preservation of microcirculation and mitochondrial redox-state. J Hepatol 2003; 38: 59–66.

60.Fernandez L, Heredia N, Grande L, Gomez Ga, Rimola A, Marco A. Preconditioning Protects Liver and Lung Damage in Rat Liver Transplantation: Role of Xanthine/Xanthine Oxidase. Hepatology 2002; 36: 562-572.

61.Yue Y, Qin Q, Cohen MV, Downey JM, Critz SD. The

relative order of mito KATP channels, free radicals and p38 MAPK in preconditioning’s protective pathway in rat heart. Cardiovasc Res 2002; 55: 681-689.

62.Tang LD, Tang ZM. Protective effects of SH-compounds on ischemia reperfusion induced arrhythmias in the isolated rat heart. Yao Xue Xue Bao 1991; 26: 91-95.

63.Forbes RA, Steenbergen CH, Murphy E. Diazoxide-induced cardioprotection requires signaling through a redox-sensitive mechanism. Circ Res 2001; 88: 802-809.

64.Hu K, Duan D, Li GR, Nattel S. Protein kinase C activates ATP-sensitive K+current in human and rabbit ventricular myocytes. Circ Res 1996; 78: 492-498.

65.Sato T, O'Rourke B, Marban E: Modulation of mitochondrial ATP-dependent K+channels by protein kinase C. Circ Res 1998; 83: 110-114.

66.Murphy E. Primary and secondary signaling pathways in early preconditioning that converge on the mitochondria to produce cardioprotection. Circ Res 2004; 94: 7-16.

67.Pain T, Yang XM, Critz SD, Yue Y, Nakano A, Liu GS, et al. Opening of mitochondrial KATP channels triggers the preconditioned state by generating free radicals. Circ Res 2000; 87: 460-466.

68.Holmuhamedov EL, Wang L, Terzic A. ATP-sensitive K+ channel openers prevent Ca++ overload in rat cardiac mitochondria. J Physiol Lond 1999; 519: 347-360.

69.Lim KH, Javadov SA, Das M, Clarke SJ, Suleiman MS, Halestrap AP. The effects of ischaemic preconditioning, diazoxide and 5-hydroxydecanoate on rat heart mitochondrial volume and respiration. J Physiol Lond 2002; 545: 961-974.

70.Shimizu S, Narita M, Tsujimito Y. Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 1999; 399: 483-487.