Direct evidence for nitric oxide stimulation of electrolyte secretion in the rat colon.
Nitric Oxide (NO) is synthesized in the intestinal tract and may serve as a physiological regulator of intestinal ion transport and/or a pathophysiologic mediator of secretory diarrhea associated with inflammatory mucosal diseases. Indirect approaches, employing inhibitors of nitric oxide synthase or compounds capable of donating NO in solution, have been used to demonstrate the effects on gastrointestinal muscle and the mucosa. To determine directly whether nitric oxide itself is capable of stimulating electrolyte secretion we mounted muscle-stripped rat distal colon in Ussing chambers and monitored short-circuit current (Isc), as an indicator of effects on mucosal ion transport. Comparisons were made to sodium nitroprusside (SNP). NO and SNP stimulated concentration-dependent (0.1 microM to 100 microM) increases in Isc, with NO being more potent than SNP. The EC50 for NO was approximately 8 microM compared to a value < 20 microM for SNP. The response to NO was immediate. In contrast, SNP required a mean lag-time of 41 +/- 4 seconds, and a significantly longer time was required for SNP to reach its maximum effect. The response to both of these agonists was blocked by bumetanide, indicating that they were stimulating a chloride ion secretory response. The cyclooxygenase inhibitor piroxicam, the neurotoxin tetrodotoxin and the inhibitor of guanylate cyclase, methylene blue, all inhibited the response to both agonists. These studies demonstrate that NO itself can stimulate chloride secretion by the rat colonic mucosa through a prostaglandin-dependent, and partially neural mechanism that may involve guanylate cyclase.
Tamai
H
H
Searle Research and Development, Skokie, IL 60077.
Gaginella
T S
TS
D000818:Animals; D002034:Bumetanide / Q000494:pharmacology; D002712:Chlorides / Q000378:metabolism*; D003106:Colon / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008751:Methylene Blue / Q000494:pharmacology; D009569:Nitric Oxide / Q000494:pharmacology*; D009599:Nitroprusside / Q000494:pharmacology; D010894:Piroxicam / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013779:Tetrodotoxin / Q000494:pharmacology
D002712:Chlorides; D002034:Bumetanide; D010894:Piroxicam; D009599:Nitroprusside; D009569:Nitric Oxide; D013779:Tetrodotoxin; D008751:Methylene Blue