Skip to main content
SpringerLink
Log in

AMP579 is revealed to be a potent A2b-adenosine receptor agonist in human 293 cells and rabbit hearts

  • Original Contribution
  • Published:
Basic Research in Cardiology Aims and scope Submit manuscript

Abstract

The mixed A1/A2a-adenosine agonist AMP579 given at reperfusion is protective in animal models of myocardial infarction. Receptor-blocking studies have indicated that the protection came from an adenosine receptor (AR), but neither A1- nor A2a-selective agonists could duplicate its protection. We recently found that A2b-selective agonists given at reperfusion are protective, and, therefore, tested whether AMP579 might also be an A2b agonist. We used human embryonic kidney cells overexpressing human A2b receptors as an assay system. In these cells, A2b receptor occupancy causes phosphorylation of ERK. AMP579 induced ERK phosphorylation with an EC50 of 250 nM and this phosphorylation could be blocked by MRS1754 or PSB1115, two highly selective blockers of human A2b receptors. We attempted to confirm our A2b hypothesis in a rabbit heart model of ischemia–reperfusion. AMP579 (500 nM) for 1 h starting at reperfusion reduced infarct size in isolated rabbit hearts exposed to 30 min of regional ischemia and 2 h of reperfusion (12.9 ± 2.2% infarction of risk zone vs. 32.0 ± 1.9% in untreated hearts). PSB1115 (500 nM) given for the first 15 min of reperfusion blocked AMP579’s protection (32.2 ± 3.1% infarction) which is consistent with an A2b mechanism. We conclude that AMP579 is a non-selective, but potent A2b-AR agonist, and that its protection against infarction is through that receptor.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Baxter GF, Hale SL, Miki T, Kloner RA, Cohen MV, Downey JM, Yellon DM (2000) Adenosine A1 agonist at reperfusion trial (AART): results of a three-center, blinded, randomized, controlled experimental infarct study. Cardiovasc Drugs Ther 14:607–614

    Article  CAS  PubMed  Google Scholar 

  2. Budde JM, Velez DA, Zhao Z-Q, Clark KL, Morris CD, Muraki S, Guyton RA, Vinten-Johansen J (2000) Comparative study of AMP579 and adenosine in inhibition of neutrophil-mediated vascular and myocardial injury during 24 h of reperfusion. Cardiovasc Res 47:294–305

    Article  CAS  PubMed  Google Scholar 

  3. Cohen MV, Downey JM (2008) Adenosine: trigger and mediator of cardioprotection. Basic Res Cardiol 103:203–215

    Article  CAS  PubMed  Google Scholar 

  4. de Zwart M, Link R, von Frijtag Drabbe Künzel JK, Cristalli G, Jacobson KA, Townsend-Nicholson A, Ijzerman AP (1998) A functional screening of adenosine analogues at the adenosine A2B receptor: a search for potent agonists. Nucleosides Nucleotides 17:969–985

    Google Scholar 

  5. Eckle T, Krahn T, Grenz A, Köhler D, Mittelbronn M, Ledent C, Jacobson MA, Osswald H, Thompson LF, Unertl K, Eltzschig HK (2007) Cardioprotection by ecto-5’-nucleotidase (CD73) and A2B adenosine receptors. Circulation 115:1581–1590

    Article  CAS  PubMed  Google Scholar 

  6. 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

    CAS  PubMed  Google Scholar 

  7. Goto M, Miura T, Iliodoromitis EK, O’Leary EL, Ishimoto R, Yellon DM, Iimura O (1991) Adenosine infusion during early reperfusion failed to limit myocardial infarct size in a collateral deficient species. Cardiovasc Res 25:943–949

    Article  CAS  PubMed  Google Scholar 

  8. Hausenloy DJ, Maddock HL, Baxter GF, Yellon DM (2002) Inhibiting mitochondrial permeability transition pore opening: a new paradigm for myocardial preconditioning? Cardiovasc Res 55:534–543

    Article  CAS  PubMed  Google Scholar 

  9. Hausenloy DJ, Tsang A, Mocanu MM, Yellon DM (2005) Ischemic preconditioning protects by activating prosurvival kinases at reperfusion. Am J Physiol 288:H971–H976

    CAS  Google Scholar 

  10. Hausenloy DJ, Yellon DM (2003) The mitochondrial permeability transition pore: its fundamental role in mediating cell death during ischaemia and reperfusion. J Mol Cell Cardiol 35:339–341

    Article  CAS  PubMed  Google Scholar 

  11. Hausenloy DJ, Yellon DM (2008) Clinical translation of cardioprotective strategies: report and recommendations of the Hatter Institute 5th International Workshop on Cardioprotection. Basic Res Cardiol 103:493–500

    Article  PubMed  Google Scholar 

  12. Kis A, Baxter GF, Yellon DM (2003) Limitation of myocardial reperfusion injury by AMP579, an adenosine A1/A2A receptor agonist: role of A2A receptor and Erk1/2. Cardiovasc Drugs Ther 17:415–425

    Article  CAS  PubMed  Google Scholar 

  13. Kopecky SL, Aviles RJ, Bell MR, Lobl JK, Tipping D, Frommell G, Ramsey K, Holland AE, Midei M, Jain A, Kellett M, Gibbons RJ (2003) A randomized, double-blinded, placebo-controlled, dose-ranging study measuring the effect of an adenosine agonist on infarct size reduction in patients undergoing primary percutaneous transluminal coronary angioplasty: The ADMIRE (AmP579 Delivery for Myocardial Infarction REduction) study. Am Heart J 146:146–152

    Article  CAS  PubMed  Google Scholar 

  14. Kuno A, Critz SD, Cui L, Solodushko V, Yang X-M, Krahn T, Albrecht B, Philipp S, Cohen MV, Downey JM (2007) Protein kinase C protects preconditioned rabbit hearts by increasing sensitivity of adenosine A2b-dependent signaling during early reperfusion. J Mol Cell Cardiol 43:262–271

    Article  CAS  PubMed  Google Scholar 

  15. Kuno A, Solenkova NV, Solodushko V, Dost T, Liu Y, Yang X-M, Cohen MV, Downey JM (2008) Infarct limitation by a protein kinase G activator at reperfusion in rabbit hearts is dependent on sensitizing the heart to A2b agonists by protein kinase C. Am J Physiol 295:H1288–H1295

    CAS  Google Scholar 

  16. Linden J, Thai T, Figler H, Jin X, Robeva AS (1999) Characterization of human A2B adenosine receptors: radioligand binding, western blotting, and coupling to Gq in human embryonic kidney 293 cells and HMC-1 mast cells. Mol Pharmacol 56:705–713

    CAS  PubMed  Google Scholar 

  17. 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

    Article  CAS  PubMed  Google Scholar 

  18. Liu GS, Thornton J, Van Winkle DM, Stanley AWH, Olsson RA, Downey JM (1991) Protection against infarction afforded by preconditioning is mediated by A1 adenosine receptors in rabbit heart. Circulation 84:350–356

    CAS  PubMed  Google Scholar 

  19. McVey MJ, Smits GJ, Cox BF, Kitzen JM, Clark KL, Perrone MH (1999) Cardiovascular pharmacology of the adenosine A1/A2-receptor agonist AMP 579: coronary hemodynamic and cardioprotective effects in the canine myocardium. J Cardiovasc Pharmacol 33:703–710

    Article  CAS  PubMed  Google Scholar 

  20. Morrison RR, Talukder MAH, Ledent C, Mustafa SJ (2002) Cardiac effects of adenosine in A2A receptor knockout hearts: uncovering A2B receptors. Am J Physiol 282:H437–H444

    CAS  Google Scholar 

  21. National Research Council (1996) Guide for the care and use of laboratory animals, 7th edn. National Academy Press, Washington, DC

    Google Scholar 

  22. Peakman M-C, Hill SJ (1994) Adenosine A2B-receptor-mediated cyclic AMP accumulation in primary rat astrocytes. Br J Pharmacol 111:191–198

    CAS  PubMed  Google Scholar 

  23. Philipp S, Yang X-M, Cui L, Davis AM, Downey JM, Cohen MV (2006) Postconditioning protects rabbit hearts through a protein kinase C-adenosine A2b receptor cascade. Cardiovasc Res 70:308–314

    Article  CAS  PubMed  Google Scholar 

  24. Smits GJ, McVey M, Cox BF, Perrone MH, Clark KL (1998) Cardioprotective effects of the novel adenosine A1/A2 receptor agonist AMP 579 in a porcine model of myocardial infarction. J Pharmacol Exp Ther 286:611–618

    CAS  PubMed  Google Scholar 

  25. Solenkova NV, Solodushko V, Cohen MV, Downey JM (2006) Endogenous adenosine protects preconditioned heart during early minutes of reperfusion by activating Akt. Am J Physiol 290:H441–H449

    CAS  Google Scholar 

  26. Thornton JD, Liu GS, Olsson RA, Downey JM (1992) Intravenous pretreatment with A1-selective adenosine analogues protects the heart against infarction. Circulation 85:659–665

    CAS  PubMed  Google Scholar 

  27. Vander Heide RS, Reimer KA (1996) Effect of adenosine therapy at reperfusion on myocardial infarct size in dogs. Cardiovasc Res 31:711–718

    Article  CAS  PubMed  Google Scholar 

  28. Xu Z, Downey JM, Cohen MV (2001) AMP 579 reduces contracture and limits infarction in rabbit heart by activating adenosine A2 receptors. J Cardiovasc Pharmacol 38:474–481

    Article  CAS  PubMed  Google Scholar 

  29. Xu Z, Downey JM, Cohen MV (2003) Timing and duration of administration are crucial for antiinfarct effect of AMP 579 infused at reperfusion in rabbit heart. Heart Dis 5:368–371

    Article  CAS  PubMed  Google Scholar 

  30. Xu Z, Yang X-M, Cohen MV, Neumann T, Heusch G, Downey JM (2000) Limitation of infarct size in rabbit hearts by the novel adenosine receptor agonist AMP 579 administered at reperfusion. J Mol Cell Cardiol 32:2339–2347

    Article  CAS  PubMed  Google Scholar 

  31. Yang X-M, Krieg T, Cui L, Downey JM, Cohen MV (2004) NECA and bradykinin at reperfusion reduce infarction in rabbit hearts by signaling through PI3 K, ERK, and NO. J Mol Cell Cardiol 36:411–421

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grant HL-20468 from the Heart, Lung, and Blood Institute of the National Institutes of Health.

Author information

Authors and Affiliations

  1. Department of Physiology, University of South Alabama College of Medicine, MSB 3074, Mobile, AL, 36688, USA

    Yanping Liu, Xiulan Yang, Xi-Ming Yang, Sheree Walker, Michael V. Cohen & James M. Downey

  2. Department of Medicine, University of South Alabama College of Medicine, Mobile, AL, USA

    Michael V. Cohen

  3. Department of Cardiology, Ernst-Moritz-Arndt University, Greifswald, Germany

    Karina Förster & Thomas Krieg

Authors
  1. Yanping Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiulan Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xi-Ming Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sheree Walker
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Karina Förster
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Michael V. Cohen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Thomas Krieg
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. James M. Downey
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to James M. Downey.

Additional information

Y. Liu and X. Yang contributed equally to this manuscript.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liu, Y., Yang, X., Yang, XM. et al. AMP579 is revealed to be a potent A2b-adenosine receptor agonist in human 293 cells and rabbit hearts. Basic Res Cardiol 105, 129–137 (2010). https://doi.org/10.1007/s00395-009-0056-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00395-009-0056-9

Keywords

Search

Navigation