title: Annexin-1 peptide Anx-1(2-26) protects adult rat cardiac myocytes from cellular injury induced by simulated ischaemia.
abstract: 1 The anti-inflammatory properties of annexin-1 peptides have been largely ascribed to their powerful antineutrophil actions in vivo. We have recently reported that the N-terminal fragment of annexin-1, Anx-1(2-26), preserves contractile function of cardiac muscle in vitro. The aim of the present study was to determine if Anx-1(2-26) elicits protective actions specifically on the cardiac myocyte (in the absence of neutrophils), using a model of metabolic inhibition to simulate ischaemia. 2 Metabolic inhibition of cardiac myocytes (4 h incubation at 37 degrees C in HEPES-containing buffer supplemented with 2-deoxy-D-glucose, D,L-lactic acid and pH adjusted to 6.5) followed by 2.5 h recovery in normal medium markedly increased creatine kinase (CK) and lactate dehydrogenase (LDH) levels by 179+/-39 and 26+/-7 IU L(-1) (both n=40, P<0.001), respectively. However, cellular injury was significantly decreased when Anx-1(2-26) (0.3 microM) was present during metabolic inhibition, CK by 74+/-10% and LDH by 71+/-6% (both n=31, P<0.001), respectively. 3 Boc 2 (10 microM), a nonselective formyl peptide receptor antagonist, present during metabolic inhibition, abolished the cardioprotective effect of Anx-1(2-26). 4 Addition of chelerythrine (10 microM), 5-hydroxydecanoate (500 microM) or SB202190 (1 microM) during metabolic inhibition also abolished Anx-1(2-26)-induced cardioprotection. 5 Cellular injury induced by metabolic inhibition was also largely prevented when myocytes were incubated with Anx-1(2-26) for 5 min with 10 min recovery prior to the insult, or when Anx-1(2-26) was present only during the recovery period following drug-free metabolic inhibition. 6 In conclusion, the annexin-1 peptide Anx-1(2-26) potently prevents cardiac myocyte injury induced by metabolic inhibition, an action that was dependent at least in part on the activation of the formyl peptide receptor family of G-protein-coupled receptors, protein kinase C, p38 mitogen-activated protein kinase and ATP-sensitive potassium channels.
authors.affiliation: Wynn Department of Metabolic Cardiology, Baker Heart Research Institute, Melbourne, Victoria, Australia. rebecca.ritchie@baker.edu.au
authors.forename: Rebecca H
authors.initials: RH
authors.lastname: Ritchie
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Gordon
authors.forename: Owen L
authors.initials: OL
authors.lastname: Woodman
authors.forename: Anh H
authors.initials: AH
authors.lastname: Cao
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Dusting
mesh_terms: D000470:Alkaloids; D000704:Analysis of Variance; D000818:Animals; D017305:Annexin A1 / Q000494:pharmacology*; D053119:Benzophenanthridines; D002478:Cells, Cultured; D003402:Creatine Kinase / Q000378:metabolism; D003652:Decanoic Acids / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006880:Hydroxy Acids / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D017202:Myocardial Ischemia / Q000517:prevention & control; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D010455:Peptides; D010617:Phenanthridines / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D011725:Pyridines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors
chemical_list: D000470:Alkaloids; D017305:Annexin A1; D053119:Benzophenanthridines; D003652:Decanoic Acids; D004791:Enzyme Inhibitors; D006880:Hydroxy Acids; D007093:Imidazoles; D010455:Peptides; D010617:Phenanthridines; D011725:Pyridines; C096153:annexin A1 peptide (2-26); C052853:5-hydroxydecanoic acid; C016299:chelerythrine; D007770:L-Lactate Dehydrogenase; D011493:Protein Kinase C; D048051:p38 Mitogen-Activated Protein Kinases; D003402:Creatine Kinase; C090942:4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole