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Effects of the thiol-oxidizing agent diamide on monochloramine-induced rat colonic electrolyte secretion



Effects of the thiol-oxidizing agent diamide on monochloramine-induced rat colonic electrolyte secretion



American Journal of Physiology 265(1 Part 1): C166-C170



The granulocyte-derived oxidant, monochloramine (NH-2Cl), is known to stimulate chloride ion secretion in rat distal colonic mucosa mounted in Ussing chambers, through mechanisms that are sensitive and insensitive to tetrodotoxin (TTX). The possible role of intracellular thiols, in the mechanism of action of NH-2Cl as a secretagogue, was evaluated with the thiol-oxidizing agent diamide and by measuring tissue sulfhydryl levels in response to NH-2Cl. Serosal exposure to the antioxidant glutathione (0.25 mM), 5 min before NH-2Cl (50 mu-M) addition, decreased the maximal effect of 50 mu-M NH-2Cl on short-circuit current (I-sc). The NH-2Cl-stimulated increase in I-sc was not affected by mucosal amiloride (5 mu-M). Pretreatment with 0.1 mM diamide shortened the lag period before the increase in I-sc in response to NH-2Cl, but it did not affect the maximal increase in I-sc. Although TTX (0.5 mu-M) increased the lag time for achievement of the maximal I-sc response to NH-2Cl, the neurotoxin did not inhibit the effect of diamide, suggesting that diamide acts primarily on the nonneural component of NH-2Cl-stimulated secretion. Incubation of colonic mucosa with NH-2Cl, with or without diamide, decreased cellular acid-soluble sulfhydryl concentrations. Taken together, the results support a role for epithelial cell thiols in NH-2Cl-stimulated electrolyte secretion by the rat colon.

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