Abstract

To determine the buffering capacity of ischemic rat myocardium, lactate production was altered by glycogen depletion prior to total global ischemia. Lactate production was monitored by ¹H-NMR spectroscopy in perfused rat hearts and determined by enzymatic assay of freeze-clamped tissue extracts. Intracellular pH was measured by ³¹P-NMR spectroscopy. The relationship between total lactate produced and pH varied considerably, depending on the final pH reached. At pH > 6.4 this relationship is linear with a total buffering capacity (Δlactate/ΔpH) of 25 μmol H⁺/g wet weight per pH unit. At lower pH values (pH < 6.4), the total buffering capacity increases progressively. Since ischemia is invariably accompanied by ATP and phosphocreatine (PCr) hydrolysis, the proton production/consumption during high-energy phosphate hydrolysis must be considered when evaluating the intrinsic buffering capacity of the myocardum against proton loads produced by lactate production from glucose and glycogen. Schemes are presented which allow an estimation of the contribution of ATP and PCr hydrolysis and the buffering by the CO₂/HCO₃⁻ system during ischemia. At pH > 6.4, the majority (about 60%) of buffering is due to hydrolysis of adenosine triphosphate, phosphocreatine in the heart, and neutralization of sodium bicarbonate in the perfusate. At pH < 6.4 an increasing proportion of cardiac buffering is from intrinsic cardiac buffers, most likely from intracellular proteins. After correction for these contributions to the observed total cardiac buffering capacity, the intrinsic buffering capacity of the myocardium can be accounted for by a high capacity (170 μmol/g wet weight) but low pK_a (5.2) buffering system.

Author Keywords: Buffering capacity; Glycogen depletion; Lactate production; Ischemia; NMR, ¹H-; NMR, ³¹P; (Rat myocardium)

Abbreviations: MDP; methylene diphosphonate

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