pHi regulation in alveolar macrophages: relative roles of Na(+)-H+ antiport and H(+)-ATPase

Abstract

In rabbit alveolar macrophages, recovery of intracellular pH (pHi) from acid loads to pHi values > or = 6.8 at an extracellular pH (pHo) of 7.4 (nominal absence of CO2-HCO3-) is insensitive to amiloride, an inhibitor of Na(+)-H+ exchange, and abolished by bafilomycin A1, an inhibitor of vacuolar-type H(+)-ATPase [A. Bidani, S.E.S. Brown, T.A. Heming, R. Gurich, and T.D. Dubose, Jr. Am. J. Physiol. 257 (Cell Physiol. 26): C65-C76, 1989; A. Bidani and S. E. S. Brown. Am. J. Physiol. 259 (Cell Physiol. 28): C586-C598, 1990]. To further evaluate the roles of Na(+)-H+ exchange and H(+)-ATPase activity in pHi regulation in rabbit alveolar macrophages, we have investigated the effects of amiloride and bafilomycin over a greater range of pHi (6.3-7.0) and pHo (5.0-7.4). The results indicate that rabbit alveolar macrophages possess H(+)-ATPase and a Na(+)-H+ antiporter, both of which are activated by decrements in pHi. However, in all cases, H(+)-ATPase activity exclusively determined basal pHi and was the principal mechanism (> 50%) for pHi recovery from intracellular acid loads. The pHi set point for activation of Na(+)-H+ exchange was approximately 6.8 at pHo of 7.4 and approximately 6.5 at pHo of 6.8. Na(+)-H+ exchange did not contribute significantly to pHi recovery at acid-loaded pHi above these set points. At pHo of 7.4 and pHi > or = 6.8, pHi recovery displayed an activation energy of approximately 11,000 kcal/mol and temperature coefficient of approximately 2.1, which are consistent with an energy-dependent process (i.e., H+ pump).(ABSTRACT TRUNCATED AT 250 WORDS)