Yanaka A, Carter K J, Lee H H, Silen W
Department of Surgery, Beth Israel Hospital, Harvard Medical School, Boston, Massachusetts.
Am J Physiol. 1990 May;258(5 Pt 1):G815-24. doi: 10.1152/ajpgi.1990.258.5.G815.
The effect of Cl- on intracellular pH (pH(i)) was studied using sheets of frog (Rana catesbeiana) fundic mucosa in which oxynticopeptic cells were selectively loaded with the acetomethoxy ester form of the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF/AM). Before the measurement of pH(i), tissues were exposed to either 10(-5) M forskolin in the serosal solution (stimulated tissues) or 3 x 10(-4) omeprazole in the serosal solution (inhibited tissues). In HCO3- and N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) buffers, pH(i) increased significantly after removal of Cl- from serosal and luminal solution, both in stimulated and inhibited tissues. The presence of Cl- in the luminal solution prevented this rise in pHi, an effect abolished by serosal 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS, 3 x 10(-4) M) but not by serosal amiloride (10(-3)M). In the presence of serosal Cl-, pH(i) increased after exposure to serosal DIDS, more prominently in the stimulated than in the inhibited tissues. These results confirm the presence of a Cl(-)-HCO3-exchanger in the basolateral membrane of oxynticopeptic cells in intact sheets of mucosa and suggest that luminal Cl- contributes to the regulation of pH(i) in oxynticopeptic cells.
使用青蛙(牛蛙)胃底黏膜片研究了氯离子对细胞内pH值(pH(i))的影响,在这些黏膜片中,泌酸-胃蛋白酶细胞被选择性地加载了pH敏感荧光染料2',7'-双(2-羧乙基)-5(6)-羧基荧光素(BCECF/AM)的乙酰甲氧基酯形式。在测量pH(i)之前,将组织暴露于浆膜溶液中的10(-5) M福斯可林(刺激组织)或浆膜溶液中的3×10(-4)奥美拉唑(抑制组织)。在碳酸氢根和N-2-羟乙基哌嗪-N'-2-乙磺酸(HEPES)缓冲液中,从浆膜和腔液中去除氯离子后,刺激组织和抑制组织中的pH(i)均显著升高。腔液中存在氯离子可阻止pH(i)的这种升高,浆膜4,4'-二异硫氰基芪-2,2'-二磺酸(DIDS,3×10(-4) M)可消除这种作用,但浆膜阿米洛利(10(-3)M)则不能。在浆膜存在氯离子的情况下,暴露于浆膜DIDS后pH(i)升高,在刺激组织中比在抑制组织中更明显。这些结果证实了完整黏膜片中泌酸-胃蛋白酶细胞基底外侧膜中存在氯离子-碳酸氢根交换体,并表明腔液中的氯离子有助于调节泌酸-胃蛋白酶细胞中的pH(i)。