Amino acids in the rat intestinal lumen regulate their own absorption from a distant intestinal site.

Intestinal nutrient transport is altered in response to changes in dietary conditions and luminal substrate level. It is not clear, however, whether an amino acid in the intestinal lumen can acutely affect its own absorption from a distant site. Our aim is to study the effect of an amino acid present in rat small intestinal segment on its own absorption from a proximal or distal site and elucidate the underlying mechanisms. The effect of instillation of alanine (Ala) in either jejunum or ileum on its own absorption at ileal or jejunal level was examined in vivo. The modulation of this intestinal regulatory loop by the following interventions was studied: tetrodotoxin (TTX) added to Ala, subdiaphragmatic vagotomy, chemical ablation of capsaicin-sensitive primary afferent (CSPA) fibers, and IV administration of calcitonin gene-related peptide (CGRP) antagonist. In addition, the kinetics of jejunal Ala absorption and the importance of Na+-dependent transport were studied in vitro after instilling Ala in the ileum. Basal jejunal Ala absorption [0.198 +/- 0.018 micromol x cm(-1) x 20 min(-1) (means +/- SD)] was significantly decreased with the instillation of 20 mM Ala in the ileum or in an adjacent distal jejunal segment (0.12 +/- 0.015; P < 0.0001 and 0.138 +/- 0.014; P < 0.002, respectively). Comparable inhibition was observed in the presence of proline in the ileum. Moreover, basal Ala absorption from the ileum (0.169 +/- 0.025) was significantly decreased by the presence of 20 mM Ala in the jejunum (0.103 +/- 0.027; P < 0.01). The inhibitory effect on jejunal Ala absorption was abolished by TTX, subdiaphragmatic vagotomy, neonatal capsaicin treatment, and CGRP antagonism. In vitro studies showed that Ala in the ileum affects Na+-mediated transport and increases K(m) without affecting Vmax. Intraluminal amino acids control their own absorption from a distant part of the intestine, by affecting the affinity of the Na+-mediated Ala transporter, through a neuronal mechanism that involves CSPA and CGRP.

Colectomy induces an aldosterone-mediated increase in jejunal glucose uptake in rats.

AIMS: The main function of the colon is water and electrolyte absorption. Total colectomy eliminates this colonic function and may alter the absorptive capacity of the small intestine for nutrients. This study examines the effect of total colectomy on jejunal glucose absorption and investigates the potential role of aldosterone in mediating the alterations in glucose uptake post-colectomy using the aldosterone antagonist spironolactone. MAIN METHODS: Total colectomy with ileo-rectal anastomosis was performed on anesthetized rats. Sham rats were identically handled without colon resection. Two days post-surgery, groups of colectomized rats were injected with either a daily subcutaneous dose of spironolactone or sesame oil for 12days. Body weight changes and food and water intake were measured in all experimental groups. Glucose absorption was measured by in-vivo single pass perfusion in the rat jejunum of control, sham, colectomized, colectomized with spironolactone, and colectomized with sesame oil treatment. Na/K ATPase, SGK1, SGLT1 and GLUT2 expressions were determined in jejunal mucosa in control, colectomized and colectomized/spironolactone injected rats by Western blot analysis. Histological assessment was performed on jejunal sections in control and colectomized groups. KEY FINDINGS: Glucose absorption significantly increased in colectomized rats with an observed increase in Na/K ATPase and SGK1 expression. No significant expression change in SGLT1 and GLUT2 was detected in the jejunum in colectomized rats. Spironolactone, however, significantly decreased the glucose uptake post-colectomy and normalized Na/K ATPase and SGK1 expression. SIGNIFICANCE: Our results suggest that jejunal glucose uptake increases post-colectomy as a possible consequence of an aldosterone-mediated function.

Localized colonic inflammation increases cytokine levels in distant small intestinal segments in the rat.

Local inflammation in the colon has been associated with nutrient malabsorption and altered motility in the small bowel. These remote effects suggest the release of mediators which can act (or alter) the function of intestinal segments located far from the primary area of inflammation. This study describes the changes in the expression of pro-inflammatory cytokines in the colon and in various segments of the small intestine in two rat models of experimental colitis. Colitis was induced by the intracolonic administration of 100 microL of 6% iodoacetamide or 250 microL of 2, 4, 6-trinitrobenzene sulfonic acid. Levels of interleukin one beta, interleukin 6, and tumor necrosis factor alpha were measured by ELISA in tissue homogenate sampled from duodenum, jejunum, ileum and colon at different time intervals. In homogenates of strips isolated from duodenum, jejunum and ileum, tumor necrosis alpha and interleukin-6, increased significantly 3-6 h after iodoacetamide or TNBS administration and remained elevated until the colonic inflammation subsided. Interleukin one beta showed comparable but delayed increase. Similar, but more pronounced increase of the three cytokines was noticed in areas of the colon adjacent to the ulcer. Histologic examinations revealed important inflammatory changes in the colon; however, examination of sections from the small intestines did not reveal significant differences between controls and rats with colitis. In conclusion, expression of pro-inflammatory cytokines is increased in remote segments of the small intestines during colitis. The findings may provide a partial explanation or a molecular substrate for the associated small bowel dysfunction.

Effect of theophylline on L-alanine absorption in the rat jejunum.

The effect of theophylline on the accumulation of L-alanine (Ala) by the rat jejunum and the mechanism involved has been investigated. Ala is rapidly accumulated by the jejunal strips in vitro and saturation is reached between 10 and 15 min. An In/Out ratio of 2.55 reflects the presence of an active component in the overall transport mechanism. Ala accumulation shows a tendency toward saturation as cellular Ala concentration increases. In the absence of Na+, Ala accumulation is reduced and a direct relationship is observed between alanine concentration in the incubation medium and its intracellular concentration. Alanine accumulation is inhibited when theophylline (TH) concentration in the incubation medium is greater than 0.5 mM. A maximum inhibition of approximately 50% in the presence of 10 mM theophylline is observed. Further inhibition (57-65%) is observed when the jejunal strips are incubated in a Na+-free medium containing 10 mM theophylline. Single-pass perfusion of the rat jejunum shows that the presence of 0.5 mM TH in the perfusate, simulating therapeutic doses, did not affect Ala absorption. However, about 55% inhibition of Ala absorption was observed when 10 mM TH was included in the perfusate. In conclusion, it could be stated that in both in vitro and in vivo studies high toxic but not therapeutic doses of TH inhibit intestinal Ala uptake. The mechanism of inhibition may be attributed to a non-carrier mediated mechanism with a minor effect noticed on Ala carrier system.

Up-regulation of nerve growth factor and interleukin-10 in inflamed and non-inflamed intestinal segments in rats with experimental colitis.

Inflammatory bowel diseases are characterized by dysregulated immune response to the normal microflora and structural and functional changes of the enteric nervous system which occur in inflamed as well as non-inflamed areas of the bowel. This study describes the changes in the expression of nerve growth factor (NGF) and interleukin-10 (IL-10) in the colon and in various segments of the small intestine in two rat models of experimental colitis induced by iodoacetamide or 2,4,6-trinitrobenzene sulfonic acid (TNBS). Levels of NGF and IL-10 were measured by ELISA in tissue homogenate sampled from duodenum, jejunum, ileum and colon at different time intervals. NGF and IL-10 increased significantly in homogenates of strips isolated from all small intestinal segments, 3-6h after iodoacetamide or TNBS administration and remained elevated until the colonic inflammation subsided. Similar but more pronounced increase occurred in areas of the colon adjacent to the ulcer. Histologic examinations revealed inflammatory changes in the colon; however, examination of sections from the small intestines did not reveal significant differences between controls and rats with colitis. The marked up-regulation of nerve growth factor and interleukin-10 in colitis suggests that they play a role in limiting or resolving inflammation and in preventing it from becoming uncontrolled. It also suggests that experimental colitis may be associated with latent inflammation in the small bowel.

Inhibitory effect of experimental colitis on fluid absorption in rat jejunum: role of the enteric nervous system, VIP, and nitric oxide.

Impairment of small intestinal absorption has been described in patients with ulcerative colitis and in animal models of experimental colitis. The pathophysiology of this dysfunction has not been elucidated. The aim of this study was to investigate the effect of chemical colitis on jejunal fluid absorption and determine the role of the enteric nervous system and some putative neurotransmitters. In a rat model of iodoacetamide-induced colitis, jejunal net fluid absorption was evaluated by the in vivo single-pass perfusion technique. The effects of 1) tetrodotoxin (TTX), 2) benzylalkonium chloride (BAC), 3) capsaicin, 4) vasoactive intestinal polypeptide (VIP) antagonism, 5) nitric oxide (NO) synthase (NOS) inhibition, and 6) 5-hydroxytryptamine type 3 and 4 (5-HT(3) and 5-HT(4)) receptor antagonism on the changes in fluid movement were investigated. A significant decrease in jejunal net fluid absorption was found 2 and 4 days after colitis induction: 26 (SD 14) and 28 (SD 19) microl x min(-1) x g dry intestinal wt(-1), respectively [P < 0.0002 compared with sham rats at 61 (SD 6.5) microl x min(-1) x g dry intestinal wt(-1)]. No histological changes were evident in jejunal sections. TTX and BAC reversed this decrease in fluid absorption: 54 (SD 13) and 44 (SD 14) microl x min(-1) x g dry intestinal wt(-1) (P = 0.0005 and P = 0.019, respectively, compared with colitis). Ablation of capsaicin-sensitive primary afferent fibers had a partial effect: 45 (SD 5) microl x min(-1) x g dry intestinal wt(-1) (P = 0.001 and P = 0.003 compared with colitis and sham, respectively). Constitutive and neuronal NOS inhibition and VIP antagonism returned jejunal net fluid absorption to normal values: 66 (SD 19), 61 (SD 5), and 56 (SD 14) microl x min(-1) x g dry intestinal wt(-1), respectively. 5-HT(3) and 5-HT(4) receptor antagonism had no effect. Chemical colitis is associated with a significant decrease in jejunal net fluid absorption. This decrease is neurally mediated and involves VIP- and NO-related mechanisms.