title: Erythropoietin protects the myocardium against reperfusion injury in vitro and in vivo.
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 microM) 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).
  
  
  CONCLUSIONS
  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.
authors.affiliation: The Hatter Institute and centre for Cardiology, University College London Hospital and Medical School, Grafton Way, London WC1E 6DB, UK.
authors.forename: A J
authors.initials: AJ
authors.lastname: Bullard
authors.forename: P
authors.initials: P
authors.lastname: Govewalla
authors.forename: D M
authors.initials: DM
authors.lastname: Yellon
mesh_terms: D000818:Animals; D004920:Erythropoiesis / Q000187:drug effects; D004921:Erythropoietin / Q000627:therapeutic use*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000502:physiology; D066298:In Vitro Techniques; D008297:Male; D009203:Myocardial Infarction / Q000188:drug therapy; D015428:Myocardial Reperfusion Injury / Q000517:prevention & control*; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D004921:Erythropoietin; D019869:Phosphatidylinositol 3-Kinases; D048049:Extracellular Signal-Regulated MAP Kinases