Effects of chloride transport inhibitors on intestinal fluid and ion transport in vivo and in vitro.
The hypothesis that intestinal fluid secretion is driven by Cl- has been tested by investigating the effects of NPPB (5-nitro-2-(3-phenyl-propylamino)-benzoate), a blocker of Cl- channels in nephrons, and the loop diuretic bumetanide, an inhibitor of the Na+, K+, 2Cl(-)-co-transporter. Both NPPB (IC50 (inhibitory concentration) approximately 100 microM) and bumetanide (IC50 approximately 2 microM) inhibited stimulated short-circuit current (Isc) in monolayers of a colonic cell line T84. NPPB also inhibited 36Cl- uptake by these cells, indicating that NPPB acts as a Cl- channel blocker in the T84 cells. NPPB (300 microM) and bumetanide (10 microM) abolished both stimulated Isc and Cl- secretion in isolated rat colonic mucosa. As judged by autoradiography, [3H]NPPB was found both in the crypts and at the surface after exposure of either side to the compound. In line with these results, NPPB and bumetanide reduced stimulated fluid secretion in everted colon sacs from the rat. In the anaesthetized rat model, neither bumetanide nor NPPB affected the net fluid transport. After luminal administration of [3H]NPPB to the rat, radioactivity was found mainly in the villus tips, whereas no labelling was found in the crypts. NPPB was bound to plasma protein (99%), and the inhibitory effects of both NPPB and bumetanide on Isc in T84 cells and fluid secretion in the colonic sacs decreased in the presence of albumin, again indicating that the compounds might not reach the in vivo target, or that the mechanism for fluid secretion in vivo may not be explained solely by the secretion of Cl-.
Fryklund
J
J
Astra Hässle AB, Mölndal, Sweden.
Mattsson
J P
JP
Berglund
M L
ML
Helander
H F
HF
Larsson
H
H
D000818:Animals; D001798:Blood Proteins / Q000378:metabolism; D001826:Body Fluids / Q000378:metabolism*; D002034:Bumetanide / Q000494:pharmacology; D002478:Cells, Cultured; D018118:Chloride Channels / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000378:metabolism*; D002713:Chlorine; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D009579:Nitrobenzoates / Q000493:pharmacokinetics / Q000494:pharmacology; D011485:Protein Binding; D011868:Radioisotopes; D051381:Rats; D017207:Rats, Sprague-Dawley; D014018:Tissue Distribution
D001798:Blood Proteins; D018118:Chloride Channels; D009579:Nitrobenzoates; D011868:Radioisotopes; D002034:Bumetanide; C058176:5-nitro-2-(3-phenylpropylamino)benzoic acid; D002713:Chlorine