Recent studies have been focused on the protective role of ischemic preconditioning against ischemia-reperfusion injury of the lung occurring following cardiopulmonary by-pass surgery and lung or heart transplantation. The present study was undertaken to investigate the role of adenosine in ischemic preconditioning in the isolated buffer-perfused rat lung. Since the pulmonary perfusion flow rate and left atrial pressure were constant, changes in pulmonary arterial pressure directly reflect changes in pulmonary vascular resistance. When compared to control values, ischemia-reperfusion injury in the form of 2 h of normothermic ischemia significantly reduced the pulmonary vasoconstrictor response to phenylephrine and KCl, increased wet-to-dry lung weight ratios and increased malondialdehyde content of rat lungs. Ischemic preconditioning in the form of one cycle of 5 min of ischemia and reperfusion applied prior to ischemia-reperfusion, as well as, adenosine preconditioning in the form of adenosine infusion prior to ischemia-reperfusion independently prevented the reduction in pulmonary vasoconstrictor responses and the increases in pulmonary edema and malondialdehyde formation in response to ischemia-reperfusion injury. Pretreatment with adenosine receptor antagonists, theophylline or 8-cyclopentyl-1,3-dipropyl xanthine (DPCPX) prior to ischemic preconditioning or adenosine preconditioning abolished the protective effects of preconditioning by ischemic preconditioning and adenosine preconditioning. The present data demonstrate that ischemic preconditioning and adenosine preconditioning prevent the vascular and biochemical alterations studied in response to ischemia-reperfusion injury in the pulmonary vascular bed of the rat. Results of the present study suggest activation of adenosine A1 receptors mediates the protective properties of ischemic preconditioning and adenosine preconditioning on ischemia-reperfusion injury in the lung. Moreover, the present data further suggest selective adenosine receptor agonists may be useful as pharmacologic preconditioning agents in preventing ischemia-reperfusion injury in lung transplantation and other forms of pulmonary vascular ischemia.