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Aim: d-myo-inositol-1,2,6-trisphosphate (alpha-trinositol, PP56), is a synthetic isomer of the intracellular second messenger, d-myo-inositol-1,4,5-trisphospahate. The pharmacological actions of alpha-trinositol include potent anti-inflammatory properties and inhibition of the secretion induced by cholera toxin and obstructive ileus. In the present study, we investigated whether alpha-trinositol was able to influence the secretion induced by heat-stable ST-toxin from Escherichia coli in the rat jejunum., Methods: A midline abdominal incision was performed in anaesthetized male Sprague-Dawley rats and a 6-7 cm long jejunal segment was isolated with intact vascular supply and placed in a chamber suspended from a force displacement transducer connected to a Grass(R) polygraph. Intestinal net fluid transport was continuously monitored gravimetrically. Crystalline ST-toxin (120 mouse units) was introduced into the intestinal lumen and left there for the rest of the experiment. When a stable secretion was observed, alpha-trinositol (60 mg kg-1 h-1) or saline were infused during 2 h, followed by a 2-h control period., Results: alpha-Trinositol induced a significant (P < 0.001) inhibition of ST-toxin secretion within 30 min, lasting until 2 h after infusion had stopped. The agent also moderately increased (P < 0.05) net fluid absorption in normal jejunum. Mean arterial pressure (P < 0.001) and heart rate (P < 0.001) were reduced by alpha-trinositol., Conclusion: The inhibition by alpha-trinositol of ST-toxin induced intestinal secretion is primarily secondary to inhibition of secretory mechanisms and only to lesser extent due to increased absorption. The detailed mechanisms of action have not been clarified but may involve suppression of inflammation possibly by means of cellular signal transduction.

Neurotensin (NT) is released from N cells in the small intestinal epithelium. Among other effects NT is known to elicit fluid secretion in the small intestine. This study was carried out in order to elucidate the mechanism by which NT elicits this secretion. Neurotensin infusions at two rates (4.5 and 45 pmol min-1 kg-1 body wt) to isolated segments of cat ileum in vivo, caused a steady rate of net fluid secretion and a release of vasoactive intestinal polypeptide into the mesenteric vein. The secretion was totally inhibited by tetrodotoxin. Hexamethonium, a nicotinic receptor antagonist, inhibited the secretion elicited by the lower but not by the higher dose of NT. Met-enkephalin also inhibited the induced secretion while pyrilamine, a histamine-I receptor antagonist had no effect. No significant change in enteric blood flow was caused by the NT infusion. These results indicate that NT elicits a nervous reflex in the enteric nervous system which, accordingly, turns the transport of the enterocytes into net fluid secretion.

In previous reports we have suggested that nervous reflexes are involved in the pathophysiology of cholera secretion and that these nervous reflexes involve a cholinergic synapse and a neuron with vasoactive intestinal polypeptide (VIP) as neurotransmitter. These proposals were further analyzed in this study. Tetrodotoxin (TTX) and lidocaine applied on the serosal surface inhibited cholera secretion in segments of rat small intestine. Fluid absorption in control rats was not significantly changed. Hexamethonium given i.v. decreased cholera secretion in the cat. No additional inhibition of cholera secretion was observed after giving TTX close i.a. Furthermore, the intestinal secretion evoked by VIP was not influenced by hexamethonium given i.v. or TTX given close i.a. The present observations support the hypothesis of a role for nervous reflexes in cholera secretion. The results suggest that at least a major part of the proposed nervous reflex(es) in cholera have a cholinergic synapse. Furthermore, the VIP-ergic neuron is situated "distal" to the cholinergic neuron in the reflex(es) closer to the effector cells.

Hydrochloric acid (0.1 M) placed on the serosal surface of a jejunal segment evoked an intestinal secretion (rats) or inhibited a net fluid absorption (cats). In rats it was demonstrated that lidocaine (placed on the serosa of periarterially denervated intestinal segments), hexamethonium (given i.v.; innervated or denervated intestines) and indomethacin (given i.v.; denervated intestines) markedly inhibited the acid induced secretion, while atropine (given i.v.) had no effect. In the cat experiments it was shown that tetrodotoxin (given close i.a. to denervated intestines) returned the rate of net fluid absorption to the control value observed before applying acid. It is concluded that exposing the intestinal serosa to an acid solution evokes a fluid secretion that is nervously mediated. Furthermore, it is proposed that prostaglandins are involved in the induction of the fluid secretion probably via a stimulation of nociceptors. It is also suggested that the results may have pathophysiological implications for some types of paralytic ileus.

Prostanoids were given intraluminally (PGE2) or infused close intra-arterially (PGE1 and PGE2) or arachidonic acid was administered intraluminally to denervated jejunal segments of the rat in vivo. These experimental manoeuvres caused a net fluid secretion, although a 1,000-fold higher concentration of the prostanoids was needed from the luminal than from the vascular side. I.v. hexamethonium or serosally applied lidocaine diminished the induced fluid secretion suggesting that the prostanoids act mainly by eliciting local secretory reflexes in the enteric nervous system. This nerve-mediated secretion is not accompanied by any increase in tissue cAMP. However, at higher i.a. concentrations of PGE2 there seems to be a non-neurogenic effect on the enterocytes associated with an increase in tissue cAMP.

Eighteen hours after intragastric inoculation Salmonella typhimurium elicited a net fluid secretion in the jejunum and ileum of rats. The mechanisms behind the secretory response were analysed in vivo. Extrinsic denervation of the experimental intestinal segments had no effect. The nerve-blocking agents hexamethonium (i.v.) and lidocaine (serosally applied) blocked the secretion but atropine had no effect. It was demonstrated that the bacteria were invasive by culturing from the intestinal wall. The presence of inflammatory reactions was supported by the facts that i.v. indomethacin blocked the secretory response and that there was an increased capillary leakage of albumin-bound Evans Blue into the interstitium of the intestinal mucosa. There was also a secretory response upon inoculation of the jejunal and ileal segments by cell-free supernatants of broth where bacteria had been cultured for 24 h and thereafter lysed. This response was, however, blocked by i.v. atropine, which had no effect on the secretion elicited by living bacteria. We could show no presence of E. coli LT, ST or cholera toxin in the cell-free supernatant by means of ELISA-tests. We therefore conclude that (1) S. typhimurium R5 invaded the rat jejunal and ileal mucosa, thereby creating an inflammatory response which in turn activated a nerve reflex(es) within the ENS leading to a net fluid secretion; (2) the presence of prostaglandins was needed for activating the reflex(es); (3) the reflex(es) contained a nicotinergic transmission; (4) enterotoxin was of no importance for the secretory response in the model used.

Duodenal HCO-3 secretion in anaesthetized rabbits was measured by continuous titration of the recirculating luminal perfusate at pH 7.4. The segment under study started 3-4 cm distal to the pylorus and was devoid of pancreatic and biliary HCO-3 secretion. On histological examination the submucosa was seen to contain Brunner's glands, mainly of a mucous type. Duodenum in rabbit secreted HCO3- at a considerably higher basal rate (100-125 mu equiv h-1 cm-1 of intestine) than has previously been found in the rat, cat or dog. The cyclo-oxygenase inhibitor indomethacin (20 mg kg-1) reduced the secretion by 30%, while prostaglandin E2 (5-80 microM, luminal) caused a dose-dependent increase. Prostaglandins thus seem to be important in regulation of duodenal HCO3- secretion in the rabbit and may play a role in duodenal protection against acid. Carbachol (1 and 10 micrograms kg-1) and atropine (0.5 and 1 mg kg-1) had no effects whereas hexamethonium (10 mg kg-1) caused a persistent decrease (25%) in secretion. Effects of electrical stimulation of the vagal nerves or injection of the alpha 2-adrenergic agonist clonidine markedly depended on the agent used for anaesthesia. In urethane-anaesthetized animals, clonidine (0.75-75 micrograms kg-1) tended to increase the secretion whereas with nembutal, clonidine (5-150 micrograms kg-1) decreased it significantly. Electrical stimulation of the cervical vagal nerves decreased the HCO3- secretion in urethane-anaesthetized animals but had no significant effect during nembutal anaesthesia. The responses in the nembutal-anaesthetized rabbit are similar to those previously observed in the cat, rat or dog.

Fresh normal rat bile premixed with cholera toxin (CT) did not significantly affect the CT-induced fluid accumulation in rat jejunal ligated loops. Bile from rats intrajejunally (i.j.) immunized three times with CT definitely inhibited CT-induced fluid secretion. Bile duct ligature (BDL) for 1-4 days in unimmunized rats, in contrast with mice, did not significantly affect subsequent CT-elicited fluid secretion in their ligated loops. BDL for 4 days in rats i.j. immunized with CT, only slightly decreased the CT-neutralizing ability of their gut loops. Passive transfer during 24 hr of bile from i.j.-immunized rats, but not from normal rats, into gut of normal recipient rats with BDL, significantly protected loops made in such recipients. The affinity-purified antibodies of immune bile, mixed with CT, neutralized its effect. Our data show that, unlike mice, rat bile acids are not required for expression of the CT effect in gut loops. In addition, bile from i.j.-immunized rats contains enough anti-CT antibodies to be protective on its own, but is not necessary for substantial gut protection against CT in i.j.-immunized BDL rats. Our results confirm a major and complementary role of both biliary and intestinal secretory IgA antibodies in protection of the rat gut mucosa against CT-induced fluid secretion.

We have analysed tissue cyclic 3'5'-adenosine monophosphate (cAMP) concentration in different fractions of the cat's small intestinal mucosa during secretion elicited in vivo by four different secretagogues: cholera toxin (administered intraluminally), vasoactive intestinal polypeptide (VIP; given i.a.), arachidonic acid (AA; administered intraluminally) and 5-hydroxytryptamine (5-HT; given i.a.). Cholera toxin was found to increase cAMP concentration in the villi but not in the crypts. The VIP, AA and 5-HT did not influence tissue cAMP concentration despite a profuse net fluid secretion. Hexamethonium inhibited secretion elicited by cholera toxin and AA but did not significantly influence tissue cAMP concentration. There is strong evidence for the view that villus and crypt regions of the small intestinal mucosa have different functions, secretion taking place in the crypts and absorption in the villi. However, the lack of cAMP increase in the crypts reported in this study suggests that cholera toxin in this model does not reach the crypts. The results are not in agreement with a role for cAMP in mediating secretion from the crypts, but are compatible with a role of cAMP in inhibiting absorption in the villi. It is suggested that the observed fluid secretion from the crypts elicited by cholera toxin, AA and 5-HT is to a major part mediated by intramural enteric reflexes.

The effect of sympathetic nerve stimulation and intra-arterial infusion of neuropeptide Y (NPY) on net fluid secretion and release of vasoactive intestinal polypeptide (VIP) was studied in the cat small intestine during a secretion due to cholera toxin. Activation of the splanchnic nerves (4 Hz, 5 ms, 5 V) decreased net fluid secretion to 57 +/- 10% of control. Concomitantly, the release of VIP was reduced to less than 50%. Furthermore, close i.a. infusion of NPY (estimated increase in plasma concentration 75 nmol l-1) reduced the net fluid secretion and VIP release to 27 +/- 5 and 28 +/- 4% of the pre-stimulatory value. The correlation between the decrease in net fluid secretion and reduction in VIP release showed a strong positive correlation (r = 0.83). These results strongly indicate that the antisecretory effect of sympathetic nerve stimulation during cholera diarrhoea is mediated by inhibition of secretory VIP neurons in the intestinal mucosa. A similar mechanism is also proposed for the intravascularly administered NPY.