Efflux of adenosine and total adenylate catabolites during alterations of the cellular energy state. An NMR study of continuous and discontinuous ischemia.

Reperfusion after continuous or discontinuous ischemia has a bearing on clinical interventions. An important question is the washout of metabolites after periods of diminished energy state of the myocardial cell. We therefore set out to determine the washout of adenosine and its metabolites after periods of ischemia in an experimental set-up which allowed non-destructive monitoring of the cellular energy state and cytosolic pH over consecutive time intervals. Isolated rat hearts were perfused with hemoglobin-free saline in a nuclear magnetic resonance spectrometer equipped for 31P NMR spectroscopy of phosphorus-containing metabolites, which could be measured over 3-min time blocks. The response of the heart when subjected to 18 min of continuous ischemia and subsequent reperfusion was compared with that when subjected to three 6-min periods of ischemia separated by 3-min periods of reperfusion. The mechanical performance of the hearts, oxygen consumption and efflux of adenosine and its metabolites were measured. The consecutive ischemic periods produced no evidence of preconditioning as judged from the cellular energy state, although the mechanical recovery was better than after continuous ischemia. During the repetitive ischemia/reperfusion protocol the efflux of adenosine was smaller, although the efflux of combined adenylate catabolites did not differ from that after continuous ischemia. The results do not support the view of adenosine being a major effector in the phenomenon of preconditioning.