A(3) adenosine receptors and mitogen-activated protein kinases in lung injury following in vivo reperfusion

Introduction Although activation of A(3) adenosine receptors attenuates reperfusion lung injury and associated apoptosis, the signaling pathway that mediates this protection remains unclear. Adenosine agonists activate mitogen-activated protein kinases, and these kinases have been implicated in ischemia/reperfusion injury; the purpose of this study was therefore to determine whether A(3) adenosine receptor stimulation with reperfusion modulates expression of the different mitogen-activated protein kinases. In addition, we compared the effect of the A(3) adenosine agonist IB-MECA with the newly synthesized, highly selective A(3) adenosine receptor agonist MRS3558 on injury in reperfused lung. Method Studies were performed in an in vivo spontaneously breathing cat model, in which the left lower lobe of the lung was isolated and subjected to 2 hours of ischemia and 3 hours of reperfusion. The selective A(3) adenosine receptor agonists IB-MECA (0.05 mg/kg, 0.1 mg/kg, or 0.3 mg/kg) and MRS3558 ( 0.05 mg/kg or 0.1 mg/kg) were administered before reperfusion. Results Both A(3) adenosine receptor agonists administered before reperfusion markedly ( P < 0.01) attenuated indices of injury and apoptosis, including the percentage of injured alveoli, wet/dry weight ratio, myeloperoxidase activity, TUNEL ( in situ TdT-mediated dUTP nick end labeling)-positive cells, and caspase 3 activity and expression. The more pronounced effects at low doses were observed with MRS3558. Increases in phosphorylated c-Jun amino-terminal protein kinase ( JNK), p38, and extracellular signal-regulated kinase (ERK) 1/2 levels were observed by the end of reperfusion compared with controls. Pretreatment with the A(3) agonists upregulated phosphorylated ERK1/2 levels but did not modify phosphorylated JNK and p38 levels. Conclusion The protective effects of A(3) adenosine receptor activation are mediated in part through upregulation of phosphorylated ERK. Also, MRS3558 was found to be more potent than IB-MECA in attenuating reperfusion lung injury. The results suggest not only that enhancement of the ERK pathway may shift the balance between cell death and survival toward cell survival, but also that A(3) agonists have potential as an effective therapy for ischemia/reperfusion-induced lung injury.