Mustonen H, Kivilaakso E
II Department of Surgery, University of Helsinki, Finland.
Gastroenterology. 1997 Sep;113(3):875-83. doi: 10.1016/s0016-5085(97)70182-3.
BACKGROUND & AIMS: The gastric mucosa must have efficient protective mechanisms to maintain physiological intracellular pH. The aim of this study was to investigate the effect of low luminal pH on apical membrane permeability.
Chambered Necturus antral mucosa was perfused with Ringer's/95% O2-5% CO2 at pH 7.25. The mucosal side was exposed to pH 4.0-2.0 with four microelectrodes placed in surface cells. Two-dimensional cable analysis was used to measure apical, basolateral, and shunt resistances. In some experiments, liquid sensor pH or Na(+)-selective microelectrodes were used.
Luminal acidification hyperpolarized apical cell membrane potential and increased apical cell membrane resistance from 21.3 +/- 2.6 (pH 7.25) to 38.0 +/- 2.3 k omega.cm2 (pH 3.0; n = 8). The increase in apical cell membrane resistance was preceded by transient intracellular acidosis from 7.32 +/- 0.07 (pH 4.0) to 7.23 +/- 0.06 (pH 3.0; n = 6). Similar intracellular acidosis (provoked by NH4+ prepulse) failed to cause the effects observed with luminal acid. The increase in apical cell membrane resistance caused by luminal acid was eliminated when N-methyl-D-glucamine+, but not Na+, was substituted for all cations in the luminal solution.
Luminal acidification (pH 3.0-2.0) closes apical amiloride-blockable Na+ channels. Protons are probably able to pass and even block these channels, but their effect in closing the channels does not occur intracellularly.
胃黏膜必须具备有效的保护机制以维持生理细胞内pH值。本研究旨在探讨低管腔pH值对顶端膜通透性的影响。
将有腔的美西螈胃窦黏膜用pH 7.25的林格氏液/95%氧气 - 5%二氧化碳灌注。在表面细胞中放置四个微电极,使黏膜侧暴露于pH 4.0 - 2.0的环境。采用二维电缆分析来测量顶端、基底外侧和旁路电阻。在一些实验中,使用了液体传感器pH或钠选择性微电极。
管腔酸化使顶端细胞膜电位超极化,并使顶端细胞膜电阻从21.3±2.6(pH 7.25)增加到38.0±2.3 kΩ·cm²(pH 3.0;n = 8)。顶端细胞膜电阻增加之前,细胞内出现短暂酸中毒,从7.32±0.07(pH 4.0)降至7.23±0.06(pH 3.0;n = 6)。类似的细胞内酸中毒(由铵预脉冲引起)未能产生管腔酸化所观察到的效应。当用N - 甲基 - D - 葡糖胺⁺而非钠离子替代管腔溶液中的所有阳离子时,管腔酸化引起的顶端细胞膜电阻增加被消除。
管腔酸化(pH 3.0 - 2.0)关闭顶端阿米洛利可阻断的钠离子通道。质子可能能够通过甚至阻断这些通道,但它们关闭通道的作用并非在细胞内发生。