Hypoxic Preconditioning of Isolated Cardiomyocytes of Adult Rat

  • Michiko Nojiri
  • Kouchi Tanonaka
  • Ken-Ichi Yabe
  • Satoshi Takeo
Part of the Progress in Experimental Cardiology book series (PREC, volume 1)


The present study was undertaken to examine whether or not cytoprotective effects of hypoxic preconditioning were detectable in isolated, quiescent cardiomyocytes of adult rats. The cardiomyocytes were incubated for 120 minutes under hypoxic condtions (sustained hypoxia), followed by 15-minute reoxygenation. Sustained hypoxia decreased the number of viable cells (from 99% to 70% of the initial cell), which consisted of rod- and square-shaped cardiomyocytes. It also decreased the number of rod-shaped cardiomyocytes (from 90% to 40% of the initial cell) and simultaneously increased the number of square-shaped cells (from 10% to 30% of the initial cell). Fifteen-minute reoxygenation resulted in a further decrease in the numbers of viable cells (less than 50% of the initial cell) and square-shaped cells (10% of the initial cell), whereas it did not change the number of rod-shaped cells. Hypoxia-reoxygenation also induced a release of purine nucleosides and bases (ATP metabolites) into the incubation medium. When the cardomyocytes were subjected to 20 minutes of hypoxic incubation, followed by 30 minutes of normoxic incubation (hypoxic preconditioning), sustained hypoxia-induced decreases in the numbers of viable cells and rod-shaped cells were attenuated (80% and 60% of the initial cell, respectively). The intervention also attenuated sustained hypoxia-induced increase in the number of square-shaped cells (18% of the initial cell). The number of rod-shaped cells subjected to hypoxic preconditioning at the end of 15-minute reoxygenation was similar to that at the end of sustained hypoxia, whereas the number of square-shaped cells decreased to 10% of the initial cells, which was similar to that of square-shaped cells without hypoxic preconditioning. The intervention also suppressed the release of ATP metabolites during hypoxia-reoxygenation.


Ischemic Precondition Hypoxic Precondition Isolate Rabbit Heart Hypoxic Incubation Sustained Hypoxia 
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  1. 1.
    Murry CE, Jennings RB, Reimer KA. 1986. Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium. Circulation 74:1124–1136.PubMedGoogle Scholar
  2. 2.
    Shiki K, Hearse DJ. 1987. Preconditioning of ischemic myocardium: reperfusion induced arrhythmias. Am J Physiol 253:H1470–H1476.PubMedGoogle Scholar
  3. 3.
    Murry CE, Richard VJ, Reimer KA, Jennings RB. 1990. Ischemic preconditioning slows energy metabolism and delays ultrastructural damage during sustained ischemic episode. Circ Res 66:913–931.PubMedGoogle Scholar
  4. 4.
    Hagar JM, Hale SL, Kloner RA. 1991. Effect of preconditioning ischemia on reperfusion arrhythmias after coronary artery occlusion and reperfusion in the rat. Circ Res 68:61–68.PubMedGoogle Scholar
  5. 5.
    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 28:1057–1061.PubMedGoogle Scholar
  6. 6.
    Armstrong S, Ganote CE. 1994. Adenosine receptor specificity in preconditioning of isolated rabbit cardiomyocytes: evidence of A3 receptor involovement. Cardiovasc Res 28:1049–1056.PubMedGoogle Scholar
  7. 7.
    Armstrong S, Downey JM, Ganote CE. 1994. Preconditioning of isolated rabbit cardiomyocytes: induction by metabolic stress and blockade by the adenosine antagonist SPT and calphostin C, a protein kinase inhibitor. Cardiovasc Res 28:72–77.PubMedGoogle Scholar
  8. 8.
    Gottlieb RA, Gruol DL, Zhu JY, Engler U. 1996. Preconditioning in rabbit cardiomyocytes. Role of pH, vacuolar proton ATPase, and apoptosis. J Clin Invest 97:2391–2398.PubMedCrossRefGoogle Scholar
  9. 9.
    Zhou X, Zhai X, Ashraf M. 1996. Direct evidence that initial oxidative stress triggered by preconditioning contributes to second window of protection by endogenous antioxidant enzyme in myocytes. Circulation 93:1177–1184.PubMedGoogle Scholar
  10. 10.
    Hayashi M, Nasa Y, Tanonaka K, Sasaki H, Miyake R, Hayashi J, Takeo S. 1995. The effects of long-term treatment with eicosapentaenoic acid and docosahexaenoic acid on hypoxia/reoxygenation injury of isolated cardiac cells in adult rats. J Mol Cell Cardiol 27:2031–2041.PubMedCrossRefGoogle Scholar
  11. 11.
    Liu J, Tanonaka K, Sanbe A, Yamamoto K, Takeo S. 1993. Beneficial effects of quinidine on post-ischemic contractile failure of isolated rat hearts. J Mol Cell Cardiol 25:1249–1263.PubMedCrossRefGoogle Scholar
  12. 12.
    Takeo S, Tanonaka K, Miyake K, Fukumoto T. 1988. Role of ATP metabolites in induction of incomplete recovery of cardiac contractile force after hypoxia. Can J Cardiol 4:193–200.PubMedGoogle Scholar
  13. 13.
    Takeo S, Tanonaka K, Miyake K, Fukumoto T. 1988. Adenine nucleotide metabolites are beneficial for recovery of cardiac contractile force after hypoxia. J Mol Cell Cardiol 20:187–199.PubMedCrossRefGoogle Scholar
  14. 14.
    Tanonaka K, Matsumoto M, Minematsu R, Miyake K, Murai R, Takeo S. 1989. Beneficial effect of amosulalol and phentolamine on post-hypoxic recovery of contractile force and energy metabolism in rabbit hearts. Br J Pharmacol 97:513–523.PubMedGoogle Scholar
  15. 15.
    Liu J, Tanonaka K, Ohtsuka Y, Sakai Y, Takeo S. 1993. Improvement of ischemia/reperfusion-induced contractile dysfunction of perfused hearts by class Ic antiarrhythmic agents. J Pharmacol Exp Ther 266:1247–1254.PubMedGoogle Scholar
  16. 16.
    Takeo S, Tanonaka K, Tazuma Y, Fukao N, Yoshikawa C, Fukumoto T, Tanaka T. 1988. Diltiazem and verapamil reduce the loss of adenine nucleotide metabolism from hypoxic hearts. J Mol Cell Cardiol 20:187–199.PubMedCrossRefGoogle Scholar
  17. 17.
    Tanonaka K, Maruyama Y, Takeo S. 1991. Beraprost, a prostacyclin mimetic agent, is beneficial for post-hypoxic recovery of cardiac function and metabolites in isolated rabbit hearts. Br J Pharmacol 104:779–786.PubMedGoogle Scholar
  18. 18.
    Schrader J, Haddy F, Gerlach E. 1977. Release of adenosine, inosine and hypoxanthine from the isolated guinea pig heart during hypoxia. Pflugers Arch 369:1–6.PubMedCrossRefGoogle Scholar
  19. 19.
    Vary TC, Angelakos ET, Schaffer SW. 1979. Relationship between adenine nucleotide metabolism and irreversible ischemic tissue damage in isolated perfused rat heart. Circ Res 45:218–225.PubMedGoogle Scholar
  20. 20.
    Takeo S, Sakanashi M. 1983. Possible mechanisms for reoxygenation-induced recovery of myocardial high-energy phosphates after hypoxia. J Mol Cell Cardiol 15:577–594.PubMedCrossRefGoogle Scholar
  21. 21.
    Takeo S, Tanonaka K, Shimizu K, Hirai K, Miyake K, Minematsu R. 1989. Beneficial effects of lidocaine and disopyramide on oxygen-deficiency-induced contractile failure and metabolic disturbance in isolated rabbit hearts. J Pharmacol Exp Ther 248:306–314.PubMedGoogle Scholar
  22. 22.
    Takeo S, Yamada H, Tanonaka K, Hayashi M, Sunagawa N. 1990. Possible involvement of membrane-stabilizing action in beneficial effect of beta adrenoceptor blocking agents on hypoxic and posthypoxic myocardium. J Pharmacol Exp Ther 254:847–856.PubMedGoogle Scholar
  23. 23.
    Fujioka H, Yoshihara S, Tanaka T, Fukumoto T, Kuroiwa A, Tanonaka K, Hayashi M, Takeo S. 1991. Enhancement of post-hypoxic contractile and metabolic recovery of perfused rat hearts by dlpropranolol: possible involvement of non-beta-receptor mediated activity. J Mol Cell Cardiol 23:949–962.PubMedCrossRefGoogle Scholar
  24. 24.
    Armstrong S, Ganote CE. 1994. Preconditioning of isolated rabbit cardiomyocytes: effects of glycolytic blockade, phorbol esters, and ischaemia. Cardiovasc Res 28:1700–1706.PubMedGoogle Scholar
  25. 25.
    Speechly-Dick ME, Mocanu MM, Yellon DM. 1994. Protein kinase C: its role in ischemic preconditioning in the rat. Circ Res 75:586–590.PubMedGoogle Scholar
  26. 26.
    Ytrehus K, Liu Y, Downey JM. 1994. Preconditioning protects ischemic rabbit heart by protein kinase C activation. Am J Physiol 266:H1145–H1152.PubMedGoogle Scholar
  27. 27.
    Li Y, Kloner RA. 1995. Does protein kinase C play a role in ischemic preconditioning in rat hearts? Am J Physiol 268:H426–H431.PubMedGoogle Scholar
  28. 28.
    Liu Y, Tsuchida A, Cohen MV, Downey JM. 1995. Pretreatment with angiotensin II activates protein kinase C and limits myocardial infarction in isolated rabbit hearts. J Mol Cell Cardiol 27:883–892.PubMedCrossRefGoogle Scholar
  29. 29.
    Mitchell MB, Meng X, Ao L, Brown JM, Harken AH, Banerjee A. 1995. Preconditioning of isolated rat heart is mediated by protein kinase C. Circ Res 76:73–81.PubMedGoogle Scholar
  30. 30.
    Goto M, Liu Y, Yang X-M, Ardell JL, Cohen MV, Downey JM. 1995. Role of bradykinin in protection of ischemic preconditioning in rabbit hearts. Circ Res 77:611–621.PubMedGoogle Scholar
  31. 31.
    Armstrong S, Ganote CE. 1995. In vitro ischaemic preconditioning of isolated rabbit cardiomyocytes: effects of selective adenosine receptor blockade and calphostin C. Cardiovasc Res 29:647–652.PubMedCrossRefGoogle Scholar
  32. 32.
    Armstrong SC, Hoover DB, Delacey MH, Ganote CE. 1996. Translocation of PKC, protein phosphatase inhibition and preconditioning of rabbit cardiomyocytes. J Mol Cell Cardiol 28:1479–1492.PubMedCrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Michiko Nojiri
    • 1
  • Kouchi Tanonaka
    • 1
  • Ken-Ichi Yabe
    • 1
  • Satoshi Takeo
    • 1
  1. 1.Tokyo University of Pharmacy and Life ScienceJapan

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