[Effect of C-terminal derivatives of sorbin on duodenal ion transports in rats]. / Effet des dérivés C-terminaux de la sorbine sur les flux ioniques duodénaux chez le rat.

OBJECTIVES AND METHODS: Synthetic derivatives of sorbin have been shown to inhibit VIP stimulated fluxes in the ileum in decreasing plasma-to-mucosa Na and Cl effluxes. The effect of this group of new peptides, without homology with any known peptides, was determined in rat duodenum where ion transport mechanisms differ. The improved technique of ligated loops in situ, was used, permitting the simultaneous measurement of net fluxes, influxes and effluxes for Na and Cl, in an integrated in vivo model. To determine the minimal active fragment of sorbin, synthetic C5, C7 and C20 peptides were tested and compared with known anti-secretor drugs such as loperamide, neuropeptide Y, somatostatine and metenkephalinamide. RESULTS: C7-sorbin was the minimal peptide able to decrease duodenal VIP-stimulated fluxes of water, Na and bicarbonate. It intervenes in increasing Na influx and more slightly Cl influx, which have been decreased by VIP. It does not modify much Na and Cl effluxes stimulated by VIP. Sorbin effect is in contrast with those of known antidiarrheic agents like somatostatine, loperamide, NPY and metenkephalinamide which chiefly decrease Cl efflux. CONCLUSIONS: Sorbin acts like an activator of absorption in the duodenum, in contrast to the other peptides or drugs and to its own anti-secretor effect in the ileum.

Pharmacokinetics and organ distribution of the sorbin C-terminal peptides.

Sorbin is a 153 amino acid peptide isolated from porcine small intestine. The heptapeptide-amide is the minimal active site of the natural molecule. A comparison of the distribution of C-7 and C-20 sorbin, which have been shown to share the activity of sorbin in increasing intestinal absorption of electrolytes, was undertaken by radioimmunoassay, after perfusion of 200 micrograms/kg/h. A longer half-life in plasma was observed for C-20 sorbin than for C-7 sorbin, with a clearance rate of 18 +/- 4 ml/min/kg vs. 40.6 +/- 13.5 ml/min/kg and a distribution volume of 192 +/- 35 ml/kg vs. 286 +/- 123 ml/kg. The accumulation of tritiated C-7 sorbin was observed in enterocytes, serosal acini of the salivary glands, and fundus chief cells. The recovery of intact peptide in the intestine was 0.06% per gram of tissue. Eighteen percent of the peptide was detected in urine.

[Comparison of hydroelectrolytic exchange characterized by calculation of saturable fluxes at three levels of the rat intestine]. / Comparaison des échanges hydroélectrolytiques caractérisés par le calcul des flux saturables à trois niveaux de l'intestin chez le rat.

Using the everted sac technique, which responded, as expected, to VIP by an increase of the secretion and to glucose by an increase of the absorption, we compared the water and electrolyte movements in the jejunum, ileum and colon in rats. Identical iso-osmolar test-solutions containing increasing NaCl concentrations, placed on the serosal and mucosal sides, allowed us to quantify fluxes in the absence of initial gradient. The measured net Na and Cl fluxes were dissociated into their two components, a passive flux from serosa to mucosa and a saturable flux from mucosa to serosa. The parameters of the saturable transport, calculated for each of the intestinal parts, showed the highest J max for the ileum (58.5 microEq.g-1.h-1 for Na and 52.8 microEq.g-1.h-1 for Cl) and the lowest Km for the colon that had the highest affinity for sodium and chloride (Km 11.1 mM for Na and 7.8 mM for Cl). These data confirm the functional difference between the three intestinal parts, with an active absorption of Na and active secretion of Cl in the jejunum, an apparent coupled Na and Cl absorption in the ileum and an active absorption with high affinity for both Na and Cl in the colon.

Impaired G-proteins and cyclic nucleotide phosphodiesterase activity in T-lymphocytes from patients with sarcoidosis.

Among the various immune abnormalities which characterize active sarcoidosis, a low proliferative response of peripheral blood lymphocytes to mitogenic lectins has long been observed. Since membrane-associated G-proteins are very likely to be crucial elements in lectin signal transduction, we investigated the binding of 5'-guanylylimidodiphosphate (GppNHp), a non hydrolyzable GTP analogue, to blood total lymphocyte membranes and to blood T-lymphocyte membranes from patients with active sarcoidosis, and from healthy control subjects. GppNHp binding was markedly decreased in peripheral cells from patients with sarcoidosis as compared to controls, suggesting the occurrence of a defect at the level of G-protein(s). A further characterization of G-proteins in these cells by means of ADP-ribose-labelling in the presence of bacterial toxins brought forward a significant decrease in the labelling of a 40 kDa protein, the major pertussis toxin substrate, in membranes from sarcoid patients, while the labelling of the major 44 kDa cholera toxin substrate proved to be unchanged with respect to control membranes. It is hypothesized that, in sarcoid lymphocytes, a defect in the negative control of adenylate cyclase mediated by the inhibitory G-protein Gi, prevents the lowering of cAMP necessary to normal mitogenic response of blood lymphocytes to stimulation. cAMP degradation by the specialized enzyme phosphodiesterase constitutes another critical step in the control of cAMP levels. Both cAMP and cGMP phosphodiesterase activities were found decreased in blood total lymphocyte preparations from sarcoid patients. With purified T-cells, although the mean cAMP and cGMP phosphodiesterase activities from sarcoid patients were found more markedly decreased with respect to healthy donors, only the decrease in cGMP phosphodiesterase was found statistically significant. The role these defects in cyclic nucleotide degradation potentially play in the disturbance of blood lymphocytes response associated with sarcoidosis is discussed.

Effect of sorbin on duodenal absorption of water and electrolytes in the rat.

Sorbin is a newly isolated intestinal peptide that has been purified because of its ability to induce water absorption. The effects that sorbin and some synthetic peptides corresponding to its C-terminal sequence have on duodenal absorption of water, chloride, and sodium were studied in comparison with the effects of vasoactive intestinal peptide (VIP), [D-Ala,Met]-enkephalinamide (DAMA), and angiotensin II. The technique of an in situ ligated duodenal loop in the rat was used for all peptides. Under the experimental conditions used, a low basal secretion of water, chloride, and sodium was obtained; VIP induced an increase of the secretion, whereas DAMA induced an absorption, both in the nanomolar dose range. Angiotensin II and sorbin induced an absorption in the picomolar dose range. The most effective doses of sorbin peptides but not of angiotensin induced the lowest final concentrations of Na+ and Cl- obtainable without inducing secondary water secretion. All synthetic peptides containing the C-terminal heptapeptide of sorbin were active in the picomolar dose range. Contrary to angiotensin, they had no effect on blood pressure.

Comparison of VIP-induced electrolyte secretion at three levels in rat small intestine.

Duodenal, jejunal and ileal loops were prepared and an iso-osmotic test solution injected, containing 80 mM Na+, 5-mM K+, 1.2 mM Ca2+, 77 mM Cl-, 10 mM HCO3- and 136 mM mannitol. 14CPEG 4000 was used as a non-absorbable marker and 36Cl was added to measure the bidirectional fluxes. During the 60-min in vivo incubation time, the duodenum actively secreted bicarbonate, a virtually zero flux in the jejunum was observed, whereas the ileum absorbed water and chloride and secreted bicarbonate. The response to the perfused doses of 0.15 to 2.4 nmol.100 g-1.h-1 of VIP (vasoactive intestinal peptide) differed qualitatively and quantitatively in the 3 segments: VIP increased bicarbonate secretion and induced chloride secretion in the duodenum, induced chloride secretion in the jejunum without changing bicarbonate minimal influx, induced bicarbonate secretion and suppressed chloride absorption in the ileum. The minimal dose required was lower in the duodenum (0.3 nmol.100 g-1.h-1) than in the jejunum and ileum (1.2 nmol.100 g-1.h-1). The functional heterogeneity of the small intestine was clearly demonstrated after VIP stimulation.