Gamma-aminobutyric acid (GABA) transport across human intestinal epithelial (Caco-2) cell monolayers

被引:62
作者
Thwaites, DT [1 ]
Basterfield, L [1 ]
McCleave, PMJ [1 ]
Carter, SM [1 ]
Simmons, NL [1 ]
机构
[1] Newcastle Univ, Sch Med, Dept Physiol Sci, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England
关键词
proton-coupled transport; GABA; amino acid; human intestine; epithelium; Caco-2; cells; cell culture; intracellular pH; short-circuit current;
D O I
10.1038/sj.bjp.0703069
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
1 Transintestinal absorption of gamma-aminobutyric acid (GABA) via a pH-dependent mechanism is demonstrated in the model human intestinal epithelial cell line Caco-2. 2 Experiments with BCECF [2',7',-bis(2-carboxyethyl)-5(6)-carboxyfluorescein]-loaded Caco-2 cells demonstrate that GABA transport across the apical membrane is coupled to proton flow into the cell. 3 Short-circuit current (I-SC) measurements using Caco-2 cell monolayers under voltage-clamped conditions demonstrate that pH-dependent GABA transport is a rheogenic process even in the absence of extracellular Na+, consistent with H+/GABA symport. 4 A range of GABA analogues were tested for their abilities to: (a) inhibit pH-dependent [H-3]GABA uptake across the apical membrane; (b) stimulate H+ flow across the apical surface of BCECF-loaded Caco-2 cell monolayers; (c) increase inward ISC across voltage-clamped Caco-2 cell monolayers. 5 Nipecotic acid, isonipecotic acid, D,L-beta-aminobutyric acid, and 3-amino-1-propanesulphonic acid each caused a marked acidification of intracellular pH and an increase in I-SC when superfused at the apical surface of Caco-2 cell monolayers. In contrast L-alpha-amino-n-butyric acid failed to induce proton flow or Ise The ability of these compounds to induce proton or current flow across the apical surface of this intestinal epithelium was closely related to the relative inhibitory effects on [H-3]GABA uptake. 6 These observations demonstrate H+/GABA symport and suggest that this transport mechanism may be accessible as a route for oral absorption of therapeutically-useful GABA analogues.
引用
收藏
页码:457 / 464
页数:8
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