Possible mechanisms for reoxygenation-induced recovery of myocardial high-energy phosphates after hypoxia.

Abstract

Changes in several factors responsible for high-energy phosphate production and metabolism in the heart perfused under hypoxic and subsequent reoxygenated conditions were studied using rabbit heart Langendorff preparation. A marked decline in myocardial ATP and creatine phosphate contents was observed with prolonged periods of hypoxia lasting from 15 to 60 min. Upon reoxygenation after 15 or 30 min hypoxia, creatine phosphate levels were fully recovered, whereas ATP contents were partially restored. Possible mechanisms responsible for reoxygenation-induced differential recovery of high-energy phosphate contents were investigated. Mitochondrial function for generating ATP was depressed upon hypoxia for longer than 15 min hypoxia, and the decreased function was found to be irreversible upon reoxygenation even after 15 min hypoxia. However, mitochondrial ability to generate ATP in the heart receiving 60 min hypoxia was still observed to some extent. Creatine phosphokinase activity of the myocardium exposed to hypoxic solution for 60 min showed only 19% depression. A release of creatine phosphokinase from the perfused heart was observed after more than 30 min of hypoxic perfusion or during reoxygenated perfusion after 60 min hypoxia. Changes in creatine phosphokinase activities of the myocardium and of the perfusate were not associated with those in myocardial high-energy phosphate contents. Hypoxia also induced significant release of adenine nucleotide metabolites from the perfused heart in a biphasic manner. Substrates responsible for the release of the metabolites were found to be mainly inosine and partly hypoxanthine. The metabolite release was also supported by our finding of a decrease in total adenine nucleotide contents of the myocardium upon hypoxia. The present results suggested a crucial role of hypoxia-induced release of adenine nucleotide metabolites in a differential recovery of ATP and creatine phosphate upon reoxygenation.