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Erythropoietin protects the myocardium against reperfusion injury in vitro and in vivo

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Abstract

Objective

Erythropoietin (EPO) is a hormone that is currently used to treat patients with renal failure and anaemia. However, it has also been shown to protect against ischaemia/reperfusion injury; this protection occurring via activation of the ERK 1/2 and PI3K pathways. Since we have previously shown activation of ERK 1/2 and PI3K to be important for protection against reperfusion–induced injury in the myocardium, this study was designed to investigate its effect in the myocardium using both an isolated perfused rat heart and an in vivo rat recovery model of ischaemia–reperfusion.

Methods

Using an in vitro isolated rat heart model of 35 minutes ischaemia and 2 hours reperfusion, EPO (50 ng/ml) was administered to the rat myocardium 5 minutes prior to reperfusion for 20 minutes. The in vivo open–chest rat model consisted of 40 minutes ischaemia followed by 24 hours reperfusion with EPO (5000 U/kg) being administered at the point of reperfusion.

Results

In the isolated perfused heart studies 50 ng/ml EPO was found to provide protection with a % I/R of 22.9% ± 6.4 vs 54.5% ± 7.4 for the ischaemic control group. To examine the mechanistic pathways involved in EPO–mediated protection, we co–administered the ERK 1/2 inhibitor, U0126 (10 uM) or the PI3K inhibitors, wortmannin, (100 nM) and LY294002 (15 µM) at reperfusion. U0126, wortmannin and LY294002 all abrogated EPO–mediated protection (% I/R 49.2% ± 5.6, 46.1% ± 5.5 and 49.9% ± 6.1 respectively, p < 0.05). In the in vivo open–chest rat model, the % I/R was significantly attenuated in EPO–treated animals from 53.6 % ± 3.7 in the control to 32.5% +/– 2.9 (p < 0.05). Likewise, wortmannin abrogated EPO–mediated protection (% I/R 50.7 ± 2.3 v EPO 32.5% ± 2.9, p < 0.05).

Conclusion

We demonstrate that EPO, administered at the point of reperfusion, reduced infarct size in an isolated perfused rat heart, in an ERK and PI3K dependent manner; in addition the mechanism was also confirmed in a whole animal model of ischaemia–reperfusion. These results suggest that EPO may be able to directly protect the myocardium against lethal reperfusion–induced injury and so offer the myocardium an additional clinical advantage over and above its ability to improve the oxygen carrying capacity of the blood.

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References

  1. Abdelrahman M, Sharples EJ, McDonald MC, Collin M, Patel NS, Yaqoob MM, Thiemermann C (2004) Erythropoietin attenuates the tissue injury associated with hemorrhagic shock and myocardial ischemia. Shock 22 (1):63–69

    Article  PubMed  Google Scholar 

  2. Bell RM, Yellon D M (2003) Bradykinin limits infarction when administered as an adjunct to reperfusion in mouse heart: the role of PI3K, Akt and eNOS. J Mol Cell Cardiol 35 (2):185–193

    PubMed  Google Scholar 

  3. Calvillo L, Latini R, Kajstura J, Leri A, Anversa P, Ghezzi P, Salio M, Cerami A, Brines M (2003) Recombinant human erythropoietin protects the myocardium from ischemia–reperfusion injury and promotes beneficial remodeling. Proc Natl Acad Sci USA 100 (8):4802–4806

    PubMed  Google Scholar 

  4. Chong ZZ, Kang JQ, Maiese K (2002) Erythropoietin is a novel vascular protectant through activation of Akt1 and mitochondrial modulation of cysteine proteases. Circulation 106 (23):2973–2979

    Article  PubMed  Google Scholar 

  5. Erbayraktar S, Grasso G, Sfacteria A, Xie QW, Coleman T, Kreilgaard M, Torup L, Sager T, Erbayraktar Z, Gokmen N, Yilmaz O, Ghezzi P, Villa P, Fratelli M, Casagrande S, Leist M, Helboe L, Gerwein J, Christensen S, Geist MA, Pedersen LO, Cerami–Hand C, Wuerth JP, Cerami A, Brines M (2003) Asialoerythropoietin is a nonerythropoietic cytokine with broad neuroprotective activity in vivo. Proc Natl Acad Sci USA 100 (11):6741–6746

    PubMed  Google Scholar 

  6. Fiordaliso F, Chimenti S, Staszewsky L, Bai A, Carlo E, Cuccovillo I, Doni M, Mengozzi M, Tonelli R, Ghezzi P, Coleman T, Brines M, Cerami A, Latini R (2005) A nonerythropoietic derivative of erythropoietin protects the myocardium from ischemia–reperfusion injury. Proc Natl Acad Sci USA 102 (6):2046–2051

    PubMed  Google Scholar 

  7. Guillard C, Chretien S, Pelus AS, Porteu F, Muller O, Mayeux P, Duprez V (2003) Activation of the mitogen–activated protein kinases Erk1/2 by erythropoietin receptor via a G(i )protein beta gammasubunit– initiated pathway. J Biol Chem 278 (13):11050–11056

    Article  PubMed  Google Scholar 

  8. Hausenloy DJ, Yellon DM (2004) New directions for protecting the heart against ischaemia–reperfusion injury: targeting the Reperfusion Injury Salvage Kinase (RISK)–pathway. Cardiovasc Res 61 (3):448–460

    Article  PubMed  Google Scholar 

  9. Junk AK, Mammis A, Savitz SI, Singh M, Roth S, Malhotra S, Rosenbaum PS, Cerami A, Brines M, Rosenbaum DM (2002) Erythropoietin administration protects retinal neurons from acute ischemia–reperfusion injury. Proc Natl Acad Sci USA 99 (16):10659–10664

    PubMed  Google Scholar 

  10. Leist M, Ghezzi P, Grasso G, Bianchi R, Villa P, Fratelli M, Savino C, Bianchi M, Nielsen J, Gerwien J, Kallunki P, Larsen AK, Helboe L, Christensen S, Pedersen LO, Nielsen M, Torup L, Sager T, Sfacteria A, Erbayraktar S, Erbayraktar Z, Gokmen N, Yilmaz O, Cerami–Hand C, Xie QW, Coleman T, Cerami A, Brines M (2004) Derivatives of erythropoietin that are tissue protective but not erythropoietic. Science 305 (5681):239–242

    Article  PubMed  Google Scholar 

  11. Lipsic E, van der Meer P, Henning RH, Suurmeijer AJ, Boddeus KM, van Veldhuisen DJ, van Gilst WH, Schoemaker RG (2004) Timing of erythropoietin treatment for cardioprotection in ischemia/ reperfusion. J Cardiovasc Pharmacol 44 (4):473–479

    Article  PubMed  Google Scholar 

  12. Mancini DM, Katz SD, Lang CC, LaManca J, Hudaihed A, Androne AS (2003) Effect of erythropoietin on exercise capacity in patients with moderate to severe chronic heart failure. Circulation 107 (2):294–299

    Article  PubMed  Google Scholar 

  13. Matsumura K, Jeremy RW, Schaper J, Becker LC (1998) Progression of myocardial necrosis during reperfusion of ischemic myocardium. Circulation 97 (8):795–804

    PubMed  Google Scholar 

  14. Michell BJ, Griffiths JE, Mitchelhill KI, Rodriguez–Crespo I, Tiganis T, Bozinovski S, de Montellano PR, Kemp BE, Pearson RB (1999) The Akt kinase signals directly to endothelial nitric oxide synthase. Curr Biol 9 (15):845–848

    Article  PubMed  Google Scholar 

  15. Moon C, Krawczyk M, Ahn D, Ahmet I, Paik D, Lakatta EG, Talan MI (2003) Erythropoietin reduces myocardial infarction and left ventricular functional decline after coronary artery ligation in rats. Proc Natl Acad Sci USA 100 (20):11612–11617

    Article  PubMed  Google Scholar 

  16. Parsa CJ, Kim J, Riel RU, Pascal LS, Thompson RB, Petrofski JA, Matsumoto A, Stamler JS, Koch WJ (2004) Cardioprotective effects of erythropoietin in the reperfused ischemic heart: a potential role for cardiac fibroblasts. J Biol Chem 279 (20):20655–20662

    Article  PubMed  Google Scholar 

  17. Parsa CJ, Matsumoto A, Kim J, Riel RU, Pascal LS, Walton GB, Thompson RB, Petrofski JA, Annex BH, Stamler JS, Koch WJ (2003) A novel protective effect of erythropoietin in the infarcted heart. J Clin Invest 112 (7):999–1007

    PubMed  Google Scholar 

  18. Rafiee P, Shi Y, Su J, Pritchard KA Jr, Tweddell JS, Baker JE (2005) Erythropoietin protects the infant heart against ischemia–reperfusion injury by triggering multiple signaling pathways. Basic Res Cardiol 100 (3):187–197

    PubMed  Google Scholar 

  19. Ruscher K, Freyer D, Karsch M, Isaev N, Megow D, Sawitzki B, Priller J, Dirnagl U, Meisel A et al. (2002) Erythropoietin is a paracrine mediator of ischemic tolerance in the brain: evidence from an in vitro model. J Neurosci 22 (23):10291–10301

    PubMed  Google Scholar 

  20. Schulman D, Latchman DS, Yellon DM (2002) Urocortin protects the heart from reperfusion injury via upregulation of p42/p44 MAPK signaling pathway. Am J Physiol Heart Circ Physiol 283 (4):H1481–H1488

    PubMed  Google Scholar 

  21. Silverberg DS, Wexler D, Blum M, Tchebiner JZ, Sheps D, Keren G, Schwartz D, Baruch R, Yachnin T, Shaked M, Schwartz I, Steinbruch S, Iaina A (2003) The effect of correction of anaemia in diabetics and non–diabetics with severe resistant congestive heart failure and chronic renal failure by subcutaneous erythropoietin and intravenous iron. Nephrol Dial Transplant 18 (1):141–146

    Article  PubMed  Google Scholar 

  22. Tramontano AF, Muniyappa R, Black AD, Blendea MC, Cohen I, Deng L, Sowers JR, Cutaia MV, El–Sherif N (2003) Erythropoietin protects cardiac myocytes from hypoxia–induced apoptosis through an Akt–dependent pathway. Biochem Biophys Res Commun 308 (4):990–994

    PubMed  Google Scholar 

  23. Vlahos CJ, Matter WF, Hui KY, Brown RF (1994) A specific inhibitor of phosphatidylinositol 3–kinase, 2–(4–morpholinyl)– 8–phenyl–4H–1–benzopyran– 4–one (LY294002). J Biol Chem 269 (7):5241–5248

    PubMed  Google Scholar 

  24. Wang X, Zhu C, Wang X, Gerwien JG, Schrattenholz A, Sandberg M, Leist M, Blomgren K (2004) The nonerythropoietic asialoerythropoietin protects against neonatal hypoxia–ischemia as potently as erythropoietin. J Neurochem 91 (4):900–910

    Article  PubMed  Google Scholar 

  25. Wright GL, Hanlon P, Amin K, Steenbergen C, Murphy E, Arcasoy MO (2004) Erythropoietin receptor expression in adult rat cardiomyocytes is associated with an acute cardioprotective effect for recombinant erythropoietin during ischemia–reperfusion injury. Faseb J 18 (9):1031–1033

    PubMed  Google Scholar 

  26. Yellon DM, Baxter GF (1999) Reperfusion injury revisited: is there a role for growth factor signaling in limiting lethal reperfusion injury? Trends Cardiovasc Med 9 (8):245–249

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. The Hatter Institute and centre for Cardiology, University College London Hospital and Medical School, Grafton Way, London WC1E 6DB, UK

    A. J. Bullard, P. Govewalla & D. M. Yellon

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  1. A. J. Bullard
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  2. P. Govewalla
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  3. D. M. Yellon
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Correspondence to D. M. Yellon.

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Bullard, A.J., Govewalla, P. & Yellon, D.M. Erythropoietin protects the myocardium against reperfusion injury in vitro and in vivo. Basic Res Cardiol 100, 397–403 (2005). https://doi.org/10.1007/s00395-005-0537-4

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  • DOI: https://doi.org/10.1007/s00395-005-0537-4

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