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The rate of release of purines (adenosine, inosine, hypoxanthine, xanthine and uric acid) from isolated working rat hearts was measured and compared to tissue concentrations of high energy phosphate compounds. Hearts were subjected to different workloads, and perfusions were performed: with normal oxygen supply (group 1); with the addition of insulin to the standard perfusion buffer, which contained glucose as energy source (group 2); in hypoxic conditions (group 3). In each group purine release increased (P less than 0.01) at higher workload and was closely related to indices of mechanical performance such as cardiac output or minute work (r = 0.902 and 0.858 in group 1, r = 0.902 and 0.851 in group 2, r = 0.851 and 0.881 in group 3, P less than 0.001 in each case). Work had no effect on adenine nucleotides but produced a significant (P less than 0.01) reduction in phosphocreatine/creatine ratio. The comparison of different groups showed that at any level of heart performance purine release was higher (P less than 0.001) in group 3 vs. group 1, and lower (P less than 0.001) in group 2 vs. group 1. High energy phosphates were reduced in group 3 vs. group 1 but were unchanged in group 2 vs. group 1. We conclude that in the isolated heart purine release is directly related to the rate of energy consumption, and inversely related to the rate of energy production. Purine release provides a sensitive method to evaluate myocardial energy metabolism, which is more sensitive than measurement of high energy phosphates.

Creatine phosphate (CP) has been shown to possess some pharmacological properties. When added to cardioplegic solutions it improves their myocardial protection. Furthermore exogenous creatine phosphate shows an anti-arrhythmic effect in the experimental animal and appears to decrease lactate formation following haemorrhagic shock. These properties have been ascribed to the conservation of the tissue stores of ATP. Recently Down et al. have observed higher rat heart levels of ATP and creatine phosphate after the intravenous administration of creatine phosphate. Since it is difficult to find the conditions for an increase of the adenylate pool, it seems of interest to establish whether the ATP increase is due to a rise of the total adenylate pool or of the energy charge or of both. Similarly the higher creatine phosphate concentration may be ascribed to a variation of the CP/creatine ratio or to an increase in the creatine plus CP pool. In the present paper we report that the administration of creatine phosphate raises both the adenylate pool and the energy charge. An increase of creatine and the creatine phosphate pool was also observed.

The influence of exogenous creatine phosphate (CP) on peroxidative heart injury was investigated in two experimental models: isolated working rat hearts and myocardial membrane preparations. In the first model the addition of 190 μ m hydrogen peroxide to the perfusion buffer caused a marked decrease of aortic flow, minute work and peak aortic pressure, and leakage of intracellular enzymes. In the presence of 10 m m CP the hemodynamic damage produced by the same concentration of hydrogen peroxide was significantly lower and enzyme release was also remarkably reduced. The protection was concentration-dependent and the whole structure of the molecule was required since creatine was found to be ineffective. In the absence of hydrogen peroxide, CP and creatine did not affect heart peformance. In microsomal membrane preparations CP decreased the formation of thiobarbituric acid-reactive material (malonaldehyde) induced by hydrogen peroxide in the presence of ferrous ions. This protection was concentration-dependent and occurred at physiological concentrations of CP. Also in this experimental model creatine had no effect and creatine plus inorganic phosphate was much less active than CP. The influence of CP on oxidative heart stress could account for the beneficial effect of this substance in different models of ischemic injury.

Cells are generally impermeable to nucleotides like ATP, ADP and AMP while nucleosides and bases readily cross the plasma membrane. A release of adenosine and of its catabolic derivatives by the myocardium of different animal species has been demonstrated in physiological and physiopathological conditions [2, 4, 13]. Furthermore there are many studies on the uptake of adenosine and inosine by the myocardial cells and their incorporation into tissue nucleotides [7–9, 15]. Taking into account the extracellular localization of adenosine receptors [5] and the role of this nucleoside as regulator of coronary blood flow and modulator of the positive inotropic effects of catecholamines [4] it was of interest to study a possible extracellular formation of adenosine from adenine nucleotides. Most mammalian tissues like muscle, liver and adipose tissue and cells like platelets, leukocytes and lymphocytes possess some enzyme activities associated with the cell surface membrane. Ecto-ATPase (EC 3.6.1.3/15) and ecto-5′-nucleotidase (EC 3.1.3.5) activities have been demonstrated in myocardial, smooth-muscle and endothelial cell membranes [3,4,8,12]. In the present paper we report a comparative study on the ability of the isolated rat heart to breakdown exogenous adenine nucleotides, nucleosides and bases: we have determined in the coronary perfusate all the degradation products in basal conditions and after perfusion with ATP, ADP, AMP, adenosine, inosine and hypoxanthine. All the perfused compounds were degraded up to uric acid that amounted to about 10%. Nucleotides were catabolized at a higher rate than nucleosides and so adenosine may accumulate outside the cells.

Transient ischemia does not induce myocardial necrosis but may be associated with prolonged contractile dysfunction ("stunned" myocardium). It has been suggested that alteration of the excitation-contraction coupling system (sarcoplasmic reticulum) could be responsible for this phenomenon. We tested this hypothesis by characterizing sarcoplasmic reticulum (SR) function in an isolated rat heart model of "stunned" myocardium (hearts reperfused after 10 min of normothermic global ischemia). At the end of the ischemic period oxalate-supported Ca-uptake was depressed either in the whole homogenate or in isolated SR (to 47% and 22% of control values, respectively). During reperfusion Ca-uptake of the whole heart homogenate recovered almost completely whereas slight but significant depression persisted in isolated SR (48 +/- 2 vs 67 +/- 4 nmol/min x mg, P less than 0.01). In the presence of ruthenium red or ryanodine, two inhibitors of SR Ca-release channels, Ca-uptake was stimulated. Both in the whole heart homogenate and in isolated SR, such stimulation was remarkably smaller after reperfusion than in control conditions (P less than 0.001) suggesting reduced conductivity state of the SR Ca-release channels. Ca-stimulated, magnesium-dependent ATPase activity was remarkably reduced during ischemia and postischemic reperfusion induced only incomplete recovery (93 +/- 18 vs 169 +/- 14 nmol ATP/min x mg protein, P less than 0.05). We conclude that complex modifications of SR function occur in the "stunned" myocardium and could contribute to the contractile impairment found in this condition.