Extracellular adenosine levels and cellular energy metabolism in ischemically preconditioned rat heart

Abstract

Objective: Microdialysis and 31P-NMR spectroscopy were used to test opposing hypotheses that ischemic preconditioning inhibits adenine nucleotide degradation and purine efflux, or that preconditioning activates cardiovascular adenosine formation to provide enhanced cardioprotection. Methods:31P-NMR spectra and matching interstitial fluid (ISF) or venous effluent samples were obtained from Langendorff perfused rat hearts. Control hearts (n=9) underwent 30 min of global normothermic ischemia and 30 min reperfusion. Preconditioned hearts (n=6) were subjected to a 5 min ischemic episode and 10 min reflow prior to 30 min ischemia and 30 min reperfusion. Effects of repetitive ischemia–reperfusion (3×5 min ischemic episodes) on adenosine levels and energy metabolism were also assessed (n=8). Results: Preconditioning improved post-ischemic recovery of heart rate x left ventricular developed pressure (71±5 vs 43±8%, P<0.05) and end-diastolic pressure (14±3 vs 29±4 mmHg, P<0.05) compared with control hearts, respectively. Preconditioning did not alter intracellular ATP, phosphocreatine (PCr), inorganic phosphate (Pi), H+ or free Mg2+ during global ischemia, but improved recoveries of PCr, Pi, and ΔGATP on reperfusion. ISF adenosine increased more than 20-fold during 30 min ischemia. The 5 min preconditioning episode increased ISF adenosine 3-fold, and reduced ISF adenosine and inosine during subsequent prolonged ischemia by up to 75%. Venous purine levels during reperfusion were also reduced by preconditioning. Accumulation of adenosine in ISF and venous effluent during repetitive ischemia was progressively reduced despite comparable changes in substrate for adenosine formation via 5′-nucleotidase (5′-AMP), and in allosteric modulators of this enzyme (Mg2+, H+, Pi, ADP, ATP). Conclusions: (i) Ischemic preconditioning reduces interstitial and vascular adenosine levels during ischemia–reperfusion, (ii) reduced ISF adenosine during ischemia is not due to reduced ischemic depletion of adenine nucleotides in preconditioned rat hearts, (iii) preconditioning may inhibit adenosine formation via 5′-nucleotidase in ischemic rat hearts, and (iv) improved functional recovery with preconditioning is unrelated to metabolic/bioenergetic changes during the ischemic insult, but may be related to improved post-ischemic recovery of [Pi] and ΔGATP in this model.