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. 1999 Jan;44(1):33–39. doi: 10.1136/gut.44.1.33

Proabsorptive and prosecretory roles for nitric oxide in cholera toxin induced secretion

J Turvill 1, F Mourad 1, M Farthing 1
PMCID: PMC1760056  PMID: 9862823

Abstract

Background—Cholera toxin causes small intestinal hypersecretion by inducing a coordinated response from enterocytes, enterochromaffin cells, enteric neurones, and the vascular supply. Nitric oxide has been implicated in the function of these separate components. 
Aims—To explore the role of nitric oxide in the totality of cholera toxin induced secretion in vivo. 
Methods—One group of adult male Wistar rats was treated with the nitric oxide synthase inhibitors NG-nitro-L-arginine methyl ester (L-NAME; subcutaneously or intraluminally), NG-methyl-L-arginine (L-NMA), or 7-nitroindazole. A second group of rats was treated with L-arginine (intraperitoneally or intraluminally) or D-arginine. The small intestine was isolated between two cannulae and instilled with 75 µg cholera toxin or saline for two hours. Small intestinal perfusion of a plasma electrolyte solution containing [14C]-PEG was undertaken to determine net water and electrolyte movement. After the experiment macroscopic and microscopic intestinal appearances were noted and jejunal 5-hydroxytryptamine concentrations were determined. 
Results—Both L-arginine and L-NAME induced secretion in the basal state, but only when administered intraluminally. Systemically applied L-NAME caused a dose dependent reduction in cholera toxin induced secretion. This was paralleled by L-NMA but not by 7-nitroindazole or by intraluminally applied L-NAME. Systemically applied L-NAME caused notable cyanosis of the intestine, consistent with mesenteric ischaemia, but no microscopic abnormalities. Systemically applied L-arginine but not D-arginine also reduced cholera toxin induced secretion and inhibited 5-hydroxytryptamine release. 
Conclusion—Nitric oxide has a duality of roles in cholera toxin induced secretion, acting both as an absorbagogue and a secretagogue. Its mechanisms of action include the maintenance of mucosal perfusion and enterochromaffin cell stabilisation. 



Keywords: cholera toxin; nitric oxide; small intestinal transport; 5-hydroxytryptamine; L-arginine; nitric oxide synthase inhibitors

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Figure 1 .

Figure 1

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Influence of systemic L-NAME (60 mg/kg), L-NMA (50 mg/kg), 7-nitroindazole (100 mg/kg), L-arginine (1 g/kg), and D-arginine (1 g/kg) and intraluminal L-NAME and L-arginine (20 mmol/l added to PES) on basal net small intestinal water movement. *p<0.0001 compared with control. 


Figure 2 .

Figure 2

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Influence of L-NAME (20, 40, and 60 mg/kg), L-NMA (50 mg/kg), and 7-nitroindazole (50 and 100 mg/kg) on cholera toxin (CT) induced net small intestinal water movement (µl/min/g dry intestinal weight). *p<0.02 compared with CT; †p<0.02 compared with CT+L-NAME 20 mg/kg; NS, CT versus CT+7-nitroindazole. 


Figure 3 .

Figure 3

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Influence of L-arginine 0.5 and 1.0 g/kg and D-arginine 1.0 g/kg on cholera toxin (CT) induced net small intestinal water movement.*p<0.002 compared with control; †p<0.0003 compared with CT+L-arginine 0.5 g/kg; NS, CT versus CT+D-arginine. 


Figure 4 .

Figure 4

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Effect of pretreatment with L-arginine or D-arginine (1 g/kg) on cholera toxin (CT) induced decrease in small intestinal 5-HT concentrations. *p<0.0001 compared with control; †p<0.0002 compared with CT and CT+D-arginine; NS, control versus CT+L-arginine. 


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