Skip to main content
SpringerLink
Log in

Cardioprotective effects of adenosine A1 and A 3 receptor activation during hypoxia in isolated rat cardiac myocytes

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Adenosine (ADO) is a well-known regulator of a variety of physiological functions in the heart. In stress conditions, like hypoxia or ischemia, the concentration of adenosine in the extracellular fluid rises dramatically, mainly through the breakdown of ATP. The degradation of adenosine in the ischemic myocytes induced damage in these cells, but it may simultaneously exert protective effects in the heart by activation of the adenosine receptors. The contribution of ADO to stimulation of protective effects was reported in human and animal hearts, but not in rat hearts. The aim of this study was to evaluate the role of adenosine A1 and A3 receptors (A1R and A3R), in protection of isolated cardiac myocytes of newborn rats from ischemic injury. The hypoxic conditions were simulated by exposure of cultured rat cardiomyocytes (4–5 days in vitro), to an atmosphere of a N2 (95%) and CO2 (5%) mixture, in glucose-free medium for 90 min. The cardiotoxic and cardioprotective effects of ADO ligands were measured by the release of lactate dehydrogenase (LDH) into the medium. Morphological investigation includes immunohistochemistry, image analysis of living and fixed cells and electron microscopy were executed. Pretreatment with the adenosine deaminase considerably increased the hypoxic damage in the cardiomyocytes indicating the importance of extracellular adenosine. Blocking adenosine receptors with selective A1 and A3 receptor antagonists abolished the protective effects of adenosine. A1R and A3R activation during the hypoxic insult delays onset of irreversible cell injury and collapse of mitochondrial membrane potential as assessed using DASPMI fluorochrom. Cardioprotection induced by the A1R agonist, CCPA, was abolished by an A1R antagonist, DPCPX, and was not affected by an A3R antagonist, MRS1523. Cardioprotection caused by the A3R agonist, Cl-IB-MECA, was antagonized completely by MRS1523 and only partially by DPCPX. Activation of both A1R and A3R together was more efficient in protection against hypoxia than by each one alone. Our study indicates that activation of either A1 or A3 adenosine receptors in the rat can attenuate myocyte injury during hypoxia. Highly selective A1R and A3R agonists may have potential as cardioprotective agents against ischemia or heart surgery.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Tucker AL, Robeva AS, Taylor HE, Holeton D, Bockner M, Lynch KR, Linden J: A1 adenosine receptors. Two amino acids are responsible for species differences in ligand recognition. J Biol Chem 269: 27900–27906, 1994

    Google Scholar 

  2. Berne RM: Cardiac nucleotides in hypoxia: Possible role in regulation of coronary blood flow. Am J Physiol 204: 317–322, 1963

    Google Scholar 

  3. Headrick JP: Ischemic preconditioning: Bioenergetic and metabolic changes and the role of endogenous adenosine. J Mol Cell Cardiol 28: 1227–1240, 1996

    Google Scholar 

  4. Wang J, Drake L, Sajjadi F, Firestein GS, Mullane KM, Bullough DA: Dual activation of adenosine A1 and A3 receptors mediates preconditioning of isolated cardiac myocytes. Eur J Pharmacol 320: 241–248, 1997

    Google Scholar 

  5. Tracey WR, Magee W, Masamune H, Kennedy SP, Knight DR, Buchholz RA, Hill RJ: Selective adenosine A3 receptor stimulation reduces ischemic myocardial injury in the rabbit heart. Cardiovasc Res 33: 410–415, 1997

    Google Scholar 

  6. Strickler J, Jacobson KA, Liang BT: Direct preconditioning of cultured chick ventricular myocytes. Novel functions of cardiac adenosine A2a and A3 receptors. J Clin Invest 98: 1773–1779, 1996

    Google Scholar 

  7. Stambaugh K, Jacobson KA, Jiang JL, Liang BT: A novel cardioprotective function of adenosine A1 and A3 receptors during prolonged simulated ischemia. Am J Physiol 273: H501–H505, 1997

    Google Scholar 

  8. Lasley RD, Narayan P, Jahania MS, Partin EL, Kraft KR, Mentzer RMJ: Species-dependent hemodynamic effects of adenosine A3-receptor agonists IB-MECA and C1-IB-MECA. Am J Physiol 276: H2076–H2084, 1999

    Google Scholar 

  9. Cave AC, Collis CS, Downey JM, Hearse DJ: Improved functional recovery by ischaemic preconditioning is not mediated by adenosine in the globally ischaemic isolated rat heart. Cardiovasc Res 27: 663–668, 1993

    Google Scholar 

  10. Nojiri M, Tanonaka K, Yabe K, Kawana K, Iwai T, Yamane M, Yoshida H, Hayashi J, Takeo S: Involvement of adenosine receptor, potassium channel and protein kinase C in hypoxic preconditioning of isolated cardiomyocytes of adult rat. Jpn J Pharmacol 80: 15–23, 1999

    Google Scholar 

  11. Shneyvays V, Nawrath H, Jacobson KA, Shainberg A: Induction of apoptosis in cardiac myocytes by an A3 adenosine receptor agonist. Exp Cell Res 243: 383–397, 1998

    Google Scholar 

  12. Shneyvays V, Jacobson KA, Li AH, Nawrath H, Zinman T, Isaac A, Shainberg A: Induction of apoptosis in rat cardiocytes by A3 adenosine receptor activation and its suppression by isoproterenol. Exp Cell Res 257: 111–126, 2000

    Google Scholar 

  13. Norton GR, Woodiwiss AJ, McGinn RJ, Lorbar M, Chung ES, Honeyman TW, Fenton RA, Dobson JG Jr, Meyer TE: Adenosine A1 receptormediated antiadrenergic effects are modulated by A2a receptor activation in rat heart. Am J Physiol 276: H341–H349, 1999

    Google Scholar 

  14. Auchampach JA, Bolli R: Adenosine receptor subtypes in the heart: Therapeutic opportunities and challenges. Am J Physiol 276: H1113–H1116, 1999

    Google Scholar 

  15. El-Ani D, Jacobson KA, Shainberg A: Characterization of adenosine receptors in intact cultured heart cells. Biochem Pharmacol 48: 727–735, 1994

    Google Scholar 

  16. Waisberg M, Shainberg A: Characterization of muscarinic cholinergic receptors in intact myocardial cells in vitro. Biochem Pharmacol 43: 2327–2334, 1992

    Google Scholar 

  17. Nieminen AL, Gores GJ, Bond JM, Imberti R, Herman B, Lemasters JJ: A novel cytotoxicity screening assay using a multiwell fluorescence scanner. Toxicol Appl Pharmacol 115: 147–155, 1992

    Google Scholar 

  18. Li AH, Moro S, Melman N, Ji XD, Jacobson KA: Structure-activity relationships and molecular modeling of 3, 5-diacyl-2,4-dialkylpyridine derivatives as selective A3 adenosine receptor antagonists. J Med Chem 41: 3186–3201, 1998

    Google Scholar 

  19. Downey JM, Liu GS, Thornton JD: Adenosine and the anti-infarct effects of preconditioning. Cardiovasc Res 27: 3–8, 1993

    Google Scholar 

  20. Jacobson KA: Adenosine A3 receptors: Novel ligands and paradoxical effects. Trends Pharmacol Sci 19: 184–191, 1998

    Google Scholar 

  21. Babbitt DG, Virmani R, Forman MB: Intracoronary adenosine administered after reperfusion limits vascular injury after prolonged ischemia in the canine model. Circulation 80: 1388–1399, 1989

    Google Scholar 

  22. Olafsson B, Forman MB, Puett DW, Pou A, Cates CU, Friesinger GC, Virmani R: Reduction of reperfusion injury in the canine preparation by intracoronary adenosine: importance of the endothelium and the noreflow phenomenon. Circulation 76: 1135–1145, 1987

    Google Scholar 

  23. Funaya H, Kitakaze M, Node K, Minamino T, Komamura K, Hori M: Plasma adenosine levels increase in patients with chronic heart failure. Circulation 95: 1363–1365, 1997

    Google Scholar 

  24. Ely SW, Berne RM: Protective effects of adenosine in myocardial ischemia. Circulation 85: 893–904, 1992

    Google Scholar 

  25. Mubagwa K, Mullane K, Flameng W: Role of adenosine in the heart circulation. Cardiovasc Res 32: 797–813, 1996

    Google Scholar 

  26. Liang BT, Jacobson KA: A physiological role of the adenosine A3 receptor: Sustained cardioprotection. Proc Natl Acad Sci USA 95: 6995–6999, 1998

    Google Scholar 

  27. Carr CS, Hill RJ, Masamune H, Kennedy SP, Knight DR, Tracey WR, Yellon DM: Evidence for a role for both the adenosine A1 and A3 receptors in protection of isolated human atrial muscle against simulated ischaemia. Cardiovasc Res 36: 52–59, 1997

    Google Scholar 

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

    Google Scholar 

  29. Kohno Y, Yoshitatsu S, Koshiba M, Kim HO, Jacobson KA: Induction of apoptosis in HL-60 human promyelocytic leukemia cells by adenosine A3 receptor agonists. Biochem Biophys Res Commun 219: 904–910, 1996

    Google Scholar 

  30. Budinger GR, Duranteau J, Chandel NS, Schumacker PT: Hibernation during hypoxia in cardiomyocytes. Role of mitochondria as the O2 sensor. J Biol Chem 273: 3320–3326, 1998

    Google Scholar 

  31. Murry CE, Jennings RB, Reimer KA: Preconditioning with ischemia: A delay of lethal cell injury in ischemic myocardium. Circulation 74: 1124–1136, 1986

    Google Scholar 

  32. Piper HM, Schwartz P, Spahr R, Hutter JF, Spieckermann PG: Absence of reoxygenation damage in isolated heart cells after anoxic injury. Pflügers Arch 401: 71–76, 1984

    Google Scholar 

  33. Ambrosio G, Zweier JL, Duilio C, Kuppusamy P, Santoro G, Elia PP, Tritto I, Cirillo P, Condorelli M, Chiariello M et al.: Evidence that mitochondrial respiration is a source of potentially toxic oxygen free radicals in intact rabbit hearts subjected to ischemia and reflow. J Biol Chem 268: 18532–18541, 1993

    Google Scholar 

  34. Olson RD, Mushlin PS: Doxorubicin cardiotoxicity: Analysis of prevailing hypotheses. FASEB J 4: 3076–3086, 1990

    Google Scholar 

  35. Maggirwar SB, Dhanraj DN, Somani SM, Ramkumar V: Adenosine acts as an endogenous activator of the cellular antioxidant defense system. Biochem Biophys Res Commun 201: 508–515, 1994

    Google Scholar 

  36. Ramkumar V, Nie Z, Rybak LP, Maggiewar SB: Adenosine, antioxidant enzymes and cytoprotection. Trends Pharmacol Sci 16: 283–285, 1995

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Gonda (Goldschmied) Medical Diagnostic Research Center, Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel

    Noam Safran, Vladimir Shneyvays, Nissim Balas & Asher Shainberg

  2. Laboratory of Bioorganic Chemistry, NIDDK, NIH, Bethesda, MD, USA

    Kenneth A. Jacobson

  3. Institute for Pharmacology, University of Mainz, Germany

    Hermann Nawrath

Authors
  1. Noam Safran
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Vladimir Shneyvays
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nissim Balas
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kenneth A. Jacobson
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hermann Nawrath
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Asher Shainberg
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Asher Shainberg.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Safran, N., Shneyvays, V., Balas, N. et al. Cardioprotective effects of adenosine A1 and A 3 receptor activation during hypoxia in isolated rat cardiac myocytes. Mol Cell Biochem 217, 143–152 (2001). https://doi.org/10.1023/A:1007209321969

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1007209321969

Search

Navigation