Marcocci C, Grollman E F
Section on Cell Regulation, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892.
Endocrinology. 1988 Oct;123(4):1705-11. doi: 10.1210/endo-123-4-1705.
In the present study we describe a Na+/H+ exchange system in FRTL-5 rat thyroid cells and its role in the regulation of intracellular pH (pHi). The pHi of these cells was measured in acidic medium (pH 6.5) using the equilibrium distribution of 14C-labeled 5,5'-dimethyl-oxazolidine-2,4-dione. The estimated pHi in 10 separate experiments was 7.30 +/- 0.01 (mean +/- SE). The pHi declined to 6.87 +/- 0.02 when equimolar concentrations of choline+ replaced Na+ in the incubation medium, but was restored when cells were placed in a medium containing 50 mM Na+. The necessity for external Na+ to maintain pHi, when cells are exposed to acidic solutions, is consistent with a Na+/H+ antiport system in these cells. This is further supported by the observation that pHi decreased to 6.70 +/- 0.01 when amiloride, an inhibitor of Na+/H+ exchange, was added, and that this agent prevented the recovery of pHi in Na+-depleted cells after addition of Na+ to the medium. This report also provides additional information concerning the relationship between H+ and I- uptake and suggests that there are two I- transport systems: one that is Na+-dependent, 4,4'-diisothiocyano 2,2'-disulfonic acid stilbene-sensitive, and not associated with intracellular acidification, and the other operative at an external pH of 6.5 and associated with intracellular acidification.
在本研究中,我们描述了FRTL-5大鼠甲状腺细胞中的一种Na⁺/H⁺交换系统及其在调节细胞内pH(pHi)中的作用。使用¹⁴C标记的5,5'-二甲基-恶唑烷-2,4-二酮的平衡分布,在酸性培养基(pH 6.5)中测量这些细胞的pHi。在10次独立实验中估计的pHi为7.30±0.01(平均值±标准误)。当在孵育培养基中用等摩尔浓度的胆碱⁺替代Na⁺时,pHi降至6.87±0.02,但当细胞置于含有50 mM Na⁺的培养基中时又恢复。当细胞暴露于酸性溶液时,外部Na⁺对维持pHi的必要性与这些细胞中的Na⁺/H⁺反向转运系统一致。当加入Na⁺/H⁺交换抑制剂氨氯吡脒时,pHi降至6.70±0.01,并且该试剂阻止了向培养基中添加Na⁺后Na⁺耗尽细胞中pHi的恢复,这一观察结果进一步支持了这一点。本报告还提供了有关H⁺与I⁻摄取之间关系的更多信息,并表明存在两种I⁻转运系统:一种是Na⁺依赖性的,对4,4'-二异硫氰基-2,2'-二磺酸芪敏感,且与细胞内酸化无关;另一种在外部pH为6.5时起作用,并与细胞内酸化有关。