Na+/H+ exchange is not operative under low-flow ischemic conditions

Using 31P-NMR the existence of Na+/H+ exchange system and its contribution to intracellular pH (pHi) regulation were examined in the isolated isovolumic rat heart under physiological and pathophysiological conditions. Ethylisopropylamiloride (EIPA) was used as a tool to search into the role of Na+/H+ exchange system. In the normal well-oxygenated heart dose-dependent negative chronotropic effects were observed with 10(-6) to 10(-5) M EIPA. After 10(-4) M the heart ceased to beat and a progressive fall of high energy phosphates compounds occurred. However, contrary to expectation pHi did not fall but rose after EIPA. In NH4Cl-loaded hearts removal of NH4Cl resulted in a fall of the pHi followed by a rapid recovery to the normal pHi. After 10(-5) M EIPA the fall of pHi became greater and there was no recovery within 35 min of observation period. This dose of EIPA, however, did not affect the time course of changes in the pHi during 60 min of low-flow ischemia (0.2 ml/min). It is concluded that pHi regulation following an acute acid loading is dependent on amiloride-sensitive Na+/H+ exchange. However, Na+/H+ exchange system does not play an important role in maintenance of the pHi under normoxic or ischemic condition. In the normoxic heart EIPA produced a decrease in heart rate without producing any change either in myocardial energy metabolism or in pHi. Thus, the compound could be categorized as a bradycardic agent.