Stimulation of colonic anion secretion by monochloramine: action sites.
During inflammatory bowel disease, reactive oxygen metabolites are released by phagocytes reacting with intraluminal NH3 to produce monochloramine (NH2Cl). NH2Cl is assumed to play role in the pathogenesis of inflammation-associated diarrhoea, as it is able to induce intestinal secretion. The aim of the present study was to determine the action sites of NH2Cl in rat colonic epithelium with Ussing chamber and fura-2 experiments. In intact mucosa, NH2Cl (5.10(-6)-10(-4) mol.l(-1)) evoked a concentration-dependent increase in short-circuit current (Isc), consistent with the induction of anion secretion, as demonstrated by anion substitution and transport blocker experiments. When the apical membrane was permeabilised by the ionophore nystatin, two basolateral action sites of NH2Cl (5.10(-5) mol.l(-1)) could be identified, i.e. an increase in the K+ conductance and a stimulation of the Na+-K+ pump. When tissues were basolaterally depolarised by a high K+ concentration, the stimulation of an apical Cl- conductance by NH2Cl was observed. In isolated colonic crypts loaded with the Ca2+-sensitive fluorescent dye fura-2, NH2Cl (5.10(-5) mol.l(-1)) evoked an increase in the intracellular Ca2+ concentration. This increase was independent from the presence of Ca2+ in the extracellular medium, but was inhibited by blockade of intracellular sarcoplasmatic, endoplasmatic Ca2+-ATPases with cyclopiazonic acid (10(-5) mol.l(-1)). The NH2Cl-evoked Ca2+ release was sensitive against inhibition of ryanodine receptors with ruthenium red (5.10(-5) mol.l(-1)) and against inhibition of inositol-1,4,5-trisphosphate (IP3) receptors with 2-aminoethoxydiphenylborate (10(-4) mol.l(-1)). Both blockers also inhibited the NH2Cl-induced increase in Isc. These results indicate that an intracellular Ca2+ release via ryanodine and/or IP3 receptors is involved in oxidant stimulation of anion secretion in rat colon.
Schultheiss
Gerhard
G
Institut für Veterinär-Physiologie, Justus-Liebig-Universität Giessen, Frankfurter Strasse 100, 35392, Giessen, Germany.
Lán Kocks
Sarah
S
Diener
Martin
M
D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D000838:Anions / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000378:metabolism; D016764:Cell Polarity / Q000187:drug effects / Q000502:physiology; D002700:Chloramines / Q000494:pharmacology*; D002712:Chlorides / Q000378:metabolism*; D003106:Colon / Q000187:drug effects* / Q000378:metabolism*; D004232:Diuretics / Q000494:pharmacology; D053496:Inositol 1,4,5-Trisphosphate Receptors; D011188:Potassium / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism
D000838:Anions; D015220:Calcium Channels; D002700:Chloramines; D002712:Chlorides; D004232:Diuretics; D053496:Inositol 1,4,5-Trisphosphate Receptors; D017382:Reactive Oxygen Species; D018160:Receptors, Cytoplasmic and Nuclear; D019837:Ryanodine Receptor Calcium Release Channel; D000643:Ammonium Chloride; D000254:Sodium-Potassium-Exchanging ATPase; C030816:chloramine; D011188:Potassium; D002118:Calcium