Basic Research in Cardiology

, Volume 91, Issue 5, pp 374–381

Loss of glycogen during preconditioning is not a prerequisite for protection of the rabbit heart

  • C. Weinbrenner
  • P. Wang
  • J. M. Downey
Original Contribution

Abstract

Depletion of gycogen has been proposed as the mechanism of protection from ischemic preconditioning. The hypothesis was tested by seeing whether pharmacological manipublation of preconditioning causes parallel changes in cardiac glycogen content. Five groups of isolated rabbit hearts were studied. Group 1 experienced 30 min of ischemia only. Group 2 (PC) was preconditioned with 5 min of global ischemia followed by 10 min of reperfusion. Group 3 was preconditioned with 5 min exposure to 400 nM bradykinin followed by a 10 min washout period. Group 4 experienced exposure to 10 μM adenosine followed by a 10 min washout period, and the fifth group was also preconditioned with 5 min ischemia and 10 min reperfusion but 100 μM8-(p-sulfophenyl) theophylline (SPT), which blocks adenosine receptors, was included in the buffer to block preconditioning's protection. Transmural biopsies were taken before treatment, just prior to the 30 min period of global ischemia, and after 30 min of global ischemia. Glycogen in the samples was digested with amyloglucosidase and the resulting glucose was assayed. Baseline glycogen averaged 17.3±0.6 μmol glucose/g wet weight. After preconditioning glycogen decreased to 13.3±1.3 μmol glucose/g wet weight (p<0.005 vs. baseline). Glycogen was similarly depleted after pharmacological preconditioning with adenosine (14.0±1.0 μmol glucose/g wet weight, p<0.05 vs. baseline) suggesting a correlation. However, when proconditioning was performed in the pressence of SPT, which blocks protection, glycogen was also depleted by the same amount (13.3±0.7 μmol glucose/g wet weight, p=ns vs. PC). Bradykinin, which also mimics preconditioning, caused no depletion of glycogen (16.3±0.8 μmol glucoseig wet weight, p=ns vs. baseline). Because preconditioning with bradykinin did not deplete glycogen and because glycogen continued to be low when protection from preconditioning was blocked with SPT, we conclude that loss of glycogen per se does not cause the protection of preconditioning.

Key words

Glycogen preconditioning adenosine SPT bradykinin 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    de Albuquerque CP, Gerstenblith G, Weiss RG (1994). Importance of metabolic inhibition and cellular pH in mediating preconditioning contractile and metabolic effects in rat hearts. Circ Res. 74:139–150Google Scholar
  2. 2.
    Asimakis GK (1996) Myocardial glycogen depletion cannot explain the cardioprotective effects of ischemic preconditioning in the rat heart. J Mol Cell Cardiol 28:563–570Google Scholar
  3. 3.
    Bøtker HE, Kimose HH, Thomassen AR, Nielsen TT (1995) Applicability of small endomyocardial biopsies for evaluation of high energy phosphates and glycogen in the heart. J Mol Cell Cardiol 27:2081–2089Google Scholar
  4. 4.
    Bøtker HE, Randsbæk F, Hansen SB Thomassen A, Nielsen TT (1995) Superiority of acid extractable glycogen for detection of metabolic changes during myocardial ischaemia. J Mol Cell Cardiol 27:1325–1332Google Scholar
  5. 5.
    Brew EC, Mitchell MB, Rehring TF, Gamboni-Robertson F, McIntyre RC, Harken AH, Banerjee A (1995) Role of bradykinin in cardiac functional protection after global ischemia-reperfusion in rat heart. Am J Physiol 269:H1370-H1378Google Scholar
  6. 6.
    Buxton DB, Fisher RA, Robertson SM, Olson MS (1987) Stimulation of glycogenolysis and vasoconstriction by adenosine and adenosine analogues in the perfused rat liver. Biochem J 248:35–41Google Scholar
  7. 7.
    Buxton DB, Robertson SM, Olson MS (1986) Stimulation of glycogenolysis by adenine nucleotides in the perfused rat liver. Biochem J 237:773–780Google Scholar
  8. 8.
    Carr RS, Neff JM (1984) Quantitative semi-automated enzymatic assay for tissue glycogen. Comp. Biochem Physiol 77B:447–449Google Scholar
  9. 9.
    Cross HR, Opie LH, Radda GK, Clarke K (1996) Is a high glycogen content beneficial or detrimental to the ischemic rat heart? A controversy resolved. Circ Res 78:482–491Google Scholar
  10. 10.
    Finegan BA, Lopaschuk GD, Coulson CS, Clanachan AS (1993) Adenosine alters glucose use during ischemia and reperfusion in isolated rat hearts. Circulation 87:900–908Google Scholar
  11. 11.
    Finegan BA, Lopaschuk GD, Gandhi M, Clanachan AS (1995) Ischemic preconditioning inhibits glycolysis and proton production in isolated working rat hearts. Am J Physiol 269:H1767-H1775Google Scholar
  12. 12.
    Goodwin GW, Taegtmeyer H (1994) Metabolic recovery of isolated working rat heart after brief global ischemia. Am J Physiol 267:H462-H470Google Scholar
  13. 13.
    Goto M, Cohen MV, Van Wylen DGL, Downey JM (1996) Attenuated purine production during subsequent ischemia in preconditioned rabbit myocardium is unrelated to the mechanism of protection. J Mol Cell Cardiol 28:447–454Google Scholar
  14. 14.
    Goto M, Liu Y, Yang XM, Ardell JL Cohen MV, Downey JM (1995) Role of bradykinin in protection of ischemic preconditioning in rabbit hearts. Circ Res 77:611–621Google Scholar
  15. 15.
    Ikonomidis JS, Shirai T, Weisel RD, Derylo B, Rao, V, Whiteside CI, Mickle DAG, Li R-K (1995) “Ischemic” or adenosine preconditioning of human ventricular cardiomyocytes is protein kinase C dependent. Circulation 92:I-12Google Scholar
  16. 16.
    Jennings RB, Reimer KA, Steenbergen C, Schaper J (1989) Total ischemia III: Effect of inhibition of anaerobic glycolysis. J Mol Cell Cardiol 21 Suppl 1:37–54Google Scholar
  17. 17.
    Lagerstrom CF, Walker WE, Taegtmeyer H (1988) Failure of glycogen depletion to improve, left ventricular function of the rabbit heart after hypothermic ischemic arrest. Circ Res 63:81–86Google Scholar
  18. 18.
    Lawson CS, Downey JM (1993) Preconditioning: state of the art myocardial protection. Cardiovasc Res 27:542–550Google Scholar
  19. 19.
    Liu GS, Richards SC, Olsson RA, Mullane K, Walsh RS, Downey JM (1994) Evidence that the adenosine A3 receptor may mediate the protection afforded by preconditioning in the isolated rabbit heart. Cardiovasc Res 28:1057–1061Google Scholar
  20. 20.
    Liu GS, Thornton JD, Van Winkle DM, Stanley AW, Olsson RA, Downey JM, (1991) Protection against infarction afforded by preconditioning, is mediated by A1 adenosine receptors in rabbit heart. Circulation 84:350–356Google Scholar
  21. 21.
    Liu Y, Downey JM (1992) Ischemic preconditioning protects against infarction in rat heart. Am J Physiol 263:H1107-H1112Google Scholar
  22. 22.
    Miura T, Iimura O (1993) Infarct size limitation by preconditioning: its phenomenological features and the key role or adenosine. Cardiovasc Res 27:36–42Google Scholar
  23. 23.
    Murry CE, Jennings RB, Reimer KA (1986) Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium. Circulation 74:1124–1136Google Scholar
  24. 24.
    Murry CE, Richard VJ, Reimer KA, Jennings RB (1990) Ischemic preconditioning slow energy metabolism and delays ultrastructural damage during a sustained ischemic episode. Circ Res 66:913–931Google Scholar
  25. 25.
    Neely JR, Grotyohann LW (1984) Role of glycolytic products in damage to ischemic myocardium. Dissociation of adenosine triphosphate levels and recovery of function of reperfused ischemic hearts. Circ Res 55:816–824Google Scholar
  26. 26.
    Nukina S, Fusaoka T, Thurman RG (1994) Glycogenolytic effect of adenosine involves ATP from hepatocytes and eicosanoids from Kupffer cells. Am J Physiol 266:G99–105Google Scholar
  27. 27.
    Parratt JR, Vegh A, Papp JG (1995) Bradykinin as an endogenous myocardial protective substance with particular reference to ischemic preconditioning—a brief review of the evidence. Can J Physiol Pharmacol 73:837–842Google Scholar
  28. 28.
    Reimer KA, Jennings RB (1986) Myocardial ischemia, hypoxial ano infarction. In: Fozzard HA, Haber E, Jennings RB, Katz AM, Morgan HE (eds) The heart and cardiovascular system. Raven Press, New York 1133–1201Google Scholar
  29. 29.
    Schaefer S, Carr LJ Prussel E, Ramasamy R (1995) Effects of glycogen depletion on ischemic injury, in isolated rat hearts: insights into preconditioning. Am J Physiol 268:H935-H944Google Scholar
  30. 30.
    Shizukuda Y, Iwamoto T, Mallet RT, Downey HF (1993) Hypoxic preconditioning attenuates stunning caused by repeated coronary artery occlusion in dog hearts. Cardiovasc Res 27:559–564Google Scholar
  31. 31.
    Tsuchida A, Liu Y, Liu GS, Cohen MV, Downey JM (1994) α1-Adrenergic agonists precondition rabbit ischemic myocardium independent of adenosine by direct activation of protein kinase C. Circ Res 75:576–585Google Scholar
  32. 32.
    Vander Heide RS, Reimer KA, Jennings RB (1993) Adenosine slows ischaemic metabolism in canine myocardium in vitro: relationship to ischaemic preconditioning. Cardiovasc Res 27:669–673Google Scholar
  33. 33.
    Vanoverschelde J-LJ, Janier MF, Bakke JE, Marshall DR, Bergmann SR (1994) Rate of glycolysis during ischemia determines extent of ischemic injury and functional recovery after reperfusion. Am J Physiol 267:H1785-H1794Google Scholar
  34. 34.
    Vanstapel F, Waebens M, Van Hecke P, Decanniere C, Stalmans W (1991) Modulation of maximal glycogenolysis in perfused rat liver by adenosine and ATP. Biochem J 277:597–602Google Scholar
  35. 35.
    Volovsek A, Subramanian R, Reboussin D (1992) Effects of duration of ischaemia during preconditioning on mechanical function, enzyme release and energy production in the isolated working rat heart. J Mol Cell Cardiol 24:1011–1019Google Scholar
  36. 36.
    Walsh RS, Borges M, Thornton JD, Cohen MV, Downey JM (1995) Hypoxia preconditions rabbit myocardium by an adenosine receptor-mediated mechanism. Can J Cardiol 11:141–146Google Scholar
  37. 37.
    Wolfe CL, Sievers RE, Visseren FLJ, Donnelly TJ (1993) Loss of myocardial protection after preconditioning correlates with the time course of glycogen recovery within the preconditioned segment. Circulation, 87:881–892Google Scholar
  38. 38.
    Woolfson RG, Patel VC, Yellon DM (1996) Pre-conditioning with adenosine leads to concentration-dependent infarct size reduction in the isolated rabbit heart. Cardiovasc Res 31:148–151Google Scholar

Copyright information

© Steinkopff Verlag 1996

Authors and Affiliations

  • C. Weinbrenner
    • 1
  • P. Wang
    • 1
  • J. M. Downey
    • 1
  1. 1.Department of PhysiologyUniversity of South Alabama College of MedicineMobileUSA

Personalised recommendations