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Am J Physiol Heart Circ Physiol 277: H28-H32, 1999;
0363-6135/99 $5.00
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Vol. 277, Issue 1, H28-H32, July 1999

Two distinct HCOminus 3-dependent H+ efflux pathways in human vascular endothelial cells

Bing Sun1,2, Richard D. Vaughan-Jones1, and Jun-Ichi Kambayashi2

1 University Laboratory of Physiology, Oxford OX1 3PT, United Kingdom; and 2 Thrombosis and Vascular Biology, Maryland Research Laboratories, Otsuka America Pharmaceutical, Rockville, Maryland 20850

Intracellular pH (pHi) regulation in human umbilical vein endothelial cells (HUVEC) was investigated. The pHi was recorded using seminaphthorhodafluor-1 (SNARF-1). Cells were intracellularly acid loaded with NH4Cl prepulse. In HEPES-buffered Tyrode (nominally HCO-3 free), pHi recovery from acid load was inhibited by 1.5 mM amiloride or Na+-free solution. Additionally, in HCO-3-buffered Tyrode, a HCO-3-dependent pHi recovery from acidosis was evident in the presence of 1.5 mM amiloride, which mediated complete recovery of pHi (7.26). In Na+-free solution, the HCO-3-dependent acid extruder mediated pHi recovery after an acid load but only back to 7.09. These results suggest that there are two HCO-3-dependent acid extruders in the HUVEC. One is Na+ dependent, and the other is Na+ independent. The former was further shown to be completely inhibited by 0.5 mM DIDS, whereas the latter was only inhibited by 24.6%. In Cl--free solution, both of the HCO-3-dependent pathways were inhibited. In conclusion, one HCO-3-dependent acid extruder in the HUVEC resembles the Na+-dependent Cl-/HCO-3 exchange found in other tissues, and the other is Cl- dependent but Na+ independent.

intracellular pH; human umbilical vein endothelial cells; seminaphthorhodafluor-1; sodium/hydrogen exchange; chloride/bicarbonate exchange


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