The effect of theophylline on the accumulation of L-alanine (Ala) by the rat jejunum and the mech... more 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.
Inflammatory bowel diseases are characterized by dysregulated immune response to the normal micro... more 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.
Intestinal nutrient transport is altered in response to changes in dietary conditions and luminal... more 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.
The effect of theophylline on the accumulation of L-alanine (Ala) by the rat jejunum and the mech... more 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.
Inflammatory bowel diseases are characterized by dysregulated immune response to the normal micro... more 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.
Intestinal nutrient transport is altered in response to changes in dietary conditions and luminal... more 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.
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Papers by Carmen Khoury