Abstract

Neutrophil-derived oxidants may be involved in inflammatory bowel disease. Stable oxidants formed by halogenation of primary amines (e.g., monochloramine, NH2Cl) are extremely potent and may be of major importance in the pathophysiological response in inflammatory bowel disease. We tested the effects of NH2Cl relative to other oxidants on muscle-stripped rat colon under short-circuit conditions. Serosal addition of the oxidants evoked a concentration-dependent increase in short-circuit current (Isc). The EC50 values were 3.2 microM for NH2Cl, 6.5 microM for HOCl and 6.5 microM for H2O2. Responses to NH2Cl and H2O2 were abolished by removal of Cl- or Ca++. Unidirectional 36Cl- flux measurements showed that both oxidants (50 microM) evoked significant decreases in net chloride absorption. Tetrodotoxin (0.5 microM) and atropine (0.5 microM) partially inhibited the initial phase of the response to 50 microM NH2Cl; tetrodotoxin completely inhibited and atropine partially inhibited the response to 50 microM H2O2. Piroxicam (5 microM) inhibited the increase in Isc to 50 microM NH2Cl and H2O2 by 20-45% and 90%, respectively. Serosal prostaglandin E2 levels were significantly increased after the addition of 50 microM NH2Cl and H2O2. The morphological response to NH2Cl consisted of changes in mucosal depth and crypt architecture. In conclusion, at concentrations found in inflamed tissue, both NH2Cl and H2O2 increase Isc probably by stimulating release of arachidonate metabolites and neurotransmitter(s). NH2Cl also may act directly on the epithelial cell to stimulate Isc and evoke Cl- secretion.