Schmid A, Scholz W, Lang H J, Popp R
Max-Planck-Institut für Biophysik, Frankfurt/Main, Germany.
Biochem Biophys Res Commun. 1992 Apr 15;184(1):112-7. doi: 10.1016/0006-291x(92)91165-m.
Na+/H+ exchange activity has been examined in endothelial cells isolated from porcine brain capillaries. Intracellular pH (pHi) changes were monitored using a confocal laser scanning microscope and the pH-sensitive fluorescence indicator 2',7'-bis-(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF). Acid load of the brain capillary endothelial cells was performed with a NH4Cl (20 mM) prepulse. In bicarbonate-free solutions pHi recovered within 3 to 10 min. Removal of extracellular Na+ ions demonstrated that H+ extrusion after an acid load of the cells was Na+ dependent. The Na+/H+ exchange could be completely blocked by EIPA (5-(N-ethyl-N-isopropyl)amiloride) as well as by the novel inhibitor 3-methylsulfonyl-4-piperidinobenzoyl guanidine hydrochloride (HOE 694) in concentrations of 1 to 10 microM, respectively. EIPA and HOE 694 in a concentration of 0.1 microM caused a partial block of Na+/H+ exchange.
已对从猪脑毛细血管分离的内皮细胞中的钠氢交换活性进行了检测。使用共聚焦激光扫描显微镜和pH敏感荧光指示剂2',7'-双-(2-羧乙基)-5,6-羧基荧光素（BCECF）监测细胞内pH（pHi）变化。用氯化铵（20 mM）预脉冲对脑毛细血管内皮细胞进行酸负荷处理。在无碳酸氢盐的溶液中，pHi在3至10分钟内恢复。去除细胞外钠离子表明，细胞酸负荷后氢离子的排出依赖于钠离子。钠氢交换可分别被浓度为1至10 microM的EIPA（5-(N-乙基-N-异丙基)氨氯吡脒）以及新型抑制剂3-甲基磺酰基-4-哌啶基苯甲酰胍盐酸盐（HOE 694）完全阻断。浓度为0.1 microM的EIPA和HOE 694会导致钠氢交换部分受阻。
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