Regulation of the PepT1 peptide transporter in the rat small intestine in response to 5-fluorouracil-induced injury.

BACKGROUND & AIMS The oligopeptide transport system of the small intestine is resistant to mucosal injury. The mechanism of this resistance was investigated by examining the activity level and expression of the peptide transporter PepT1 in the intestine of rats treated with 5-fluorouracil. METHODS The expression and localization of PepT1 were examined by immunoblot analysis of brush border membrane vesicles and immunohistochemical analysis of intestinal sections with PepT1-specific rabbit polyclonal antibodies. Also, Northern blot analysis was used for the expression of PepT1 messenger RNA (mRNA). RESULTS Although the amounts of sucrase and an Na+-dependent glucose transporter protein in intestinal vesicles decreased markedly after 5-fluorouracil treatment, the amount of PepT1 protein remained largely unaffected. Immunohistochemical analysis also showed that the PepT1 immunoreactivity level was preserved in the brush border membrane of the remaining villi of 5-fluorouracil-treated rats. Levels of amino acid, glucose, and phosphate transporter mRNAs were profoundly depressed in 5-fluorouracil-treated animals, whereas the level of PepT1 mRNA conversely increased. CONCLUSIONS The resistance of intestinal peptide transport to tissue injury may be attributable to increased synthesis of PepT1 rather than to a change in the kinetic properties of the residual absorbing cells.

[1]  Y. Fukui,et al.  The basic amino acid transporter (rBAT) -like immunoreactivity in paraventricular and supraoptic magnocellular neurons of the rat hypothalamus , 1996, Brain Research.

[2]  V. Ganapathy,et al.  Expression and protein kinase C-dependent regulation of peptide/H+ co-transport system in the Caco-2 human colon carcinoma cell line. , 1994, The Biochemical journal.

[3]  L. Johnson,et al.  Physiology of the gastrointestinal tract , 2012 .

[4]  V. Ganapathy,et al.  H(+)-peptide cotransport in Madin-Darby canine kidney cells: expression and calmodulin-dependent regulation. , 1995, The American journal of physiology.

[5]  H. Schedl,et al.  Diglycine absorption in streptozotocin diabetic rat. , 1978, The American journal of physiology.

[6]  M. Romero,et al.  Expression cloning of a mammalian proton-coupled oligopeptide transporter , 1994, Nature.

[7]  M. Hediger,et al.  Human Intestinal H+/Peptide Cotransporter , 1995, The Journal of Biological Chemistry.

[8]  H. Higashida,et al.  Functional expression of transporter for beta-lactam antibiotics and dipeptides in Xenopus laevis oocytes injected with messenger RNA from human, rat and rabbit small intestines. , 1995, The Journal of pharmacology and experimental therapeutics.

[9]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Levin Rj.  Anatomical and functional changes of the small intestine induced by 5-fluorouracil. , 1968, The Journal of physiology.

[11]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[12]  V. Ganapathy,et al.  Identification of the histidyl residue obligatory for the catalytic activity of the human H+/peptide cotransporters PEPT1 and PEPT2. , 1997, Biochemistry.

[13]  K. Miyamoto,et al.  Role of liver‐type glucose transporter (GLUT2) in transport across the basolateral membrane in rat jejunum , 1992, FEBS letters.

[14]  J. Lokich,et al.  Sequential infusional 5‐fluorouracil followed by concomitant radiation for tumors of the esophagus and gastroesophageal junction , 1987, Cancer.

[15]  D. Matthews,et al.  Effects of dietary restriction and protein deprivation on intestinal absorption of protein digestion products in the rat. , 1972, The British journal of nutrition.

[16]  H. Schedl,et al.  Intestinal adaptation in diabetes: amino acid absorption. , 1974, American Journal of Physiology.

[17]  M. Lindstrom,et al.  Regional expression and dietary regulation of rat small intestinal peptide and amino acid transporter mRNAs. , 1995, Biochemical and biophysical research communications.

[18]  S. Adibi,et al.  Absorptive and digestive function of the jejunum after jejunoileal bypass for treatment of human obesity. , 1976, Gastroenterology.

[19]  M. Hediger,et al.  The neuronal and epithelial human high affinity glutamate transporter. Insights into structure and mechanism of transport. , 1994, The Journal of biological chemistry.

[20]  H. Saito,et al.  Cloning and characterization of a rat H+/peptide cotransporter mediating absorption of beta-lactam antibiotics in the intestine and kidney. , 1995, The Journal of pharmacology and experimental therapeutics.

[21]  Y. Misumi,et al.  Primary structure of rat liver dipeptidyl peptidase IV deduced from its cDNA and identification of the NH2-terminal signal sequence as the membrane-anchoring domain. , 1989, The Journal of biological chemistry.

[22]  P. Traber.  Regulation of sucrase-isomaltase gene expression along the crypt-villus axis of rat small intestine. , 1990, Biochemical and biophysical research communications.

[23]  A. Dahlqvist.  Assay of intestinal disaccharidases. , 1984, Scandinavian journal of clinical and laboratory investigation.

[24]  G. Bounous,et al.  Elemental diet in the management of the intestinal lesion produced by 5-fluorouracil in man. , 1971, Canadian journal of surgery. Journal canadien de chirurgie.

[25]  T. Nakanishi,et al.  Immunolocalization and pharmacological relevance of oligopeptide transporter PepT1 in intestinal absorption of β‐lactam antibiotics , 1996, FEBS letters.

[26]  D. Silk,et al.  Peptides in Human Nutrition , 1989, Nutrition Research Reviews.

[27]  H. Yamamoto,et al.  Sequence, tissue distribution and developmental changes in rat intestinal oligopeptide transporter. , 1996, Biochimica et biophysica acta.

[28]  Daniel G. Miller,et al.  Cell-surface receptors for gibbon ape leukemia virus and amphotropic murine retrovirus are inducible sodium-dependent phosphate symporters. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[29]  K. Miyamoto,et al.  Differential responses of intestinal glucose transporter mRNA transcripts to levels of dietary sugars. , 1993, The Biochemical journal.

[30]  P. Houghton,et al.  Mechanism of induction of gastrointestinal toxicity in the mouse by 5-fluorouracil, 5-fluorouridine, and 5-fluoro-2'-deoxyuridine. , 1979, Cancer research.

[31]  G. Semenza,et al.  A modified procedure for the rapid preparation of efficiently transporting vesicles from small intestinal brush border membranes. Their use in investigating some properties of D-glucose and choline transport systems. , 1978, Biochimica et biophysica acta.

[32]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[33]  H. Saito,et al.  Identification of the histidine residues involved in substrate recognition by a rat H+/peptide cotransporter, PEPT1 , 1996, FEBS letters.

[34]  K. Miyamoto,et al.  Inhibition of glucose absorption by phlorizin affects intestinal functions in rats. , 1993, Gastroenterology.

[35]  H. Yamamoto,et al.  Effects of Truncation of the COOH-terminal Region of a Na+-independent Neutral and Basic Amino Acid Transporter on Amino Acid Transport in Xenopus Oocytes* , 1996, The Journal of Biological Chemistry.

[36]  M. L. Gardner,et al.  Changes in absorptive and peptide hydrolase activities in rat small intestine after administration of 5-fluorouracil. , 1978, Clinical science and molecular medicine.

[37]  S. Adibi,et al.  Comparison of free amino acid and dipeptide absorption in the jejunum of sprue patients. , 1974, Gastroenterology.

[38]  M. Fukushima,et al.  Inhibition by oxonic acid of gastrointestinal toxicity of 5-fluorouracil without loss of its antitumor activity in rats. , 1993, Cancer research.

[39]  J. Bernier,et al.  Correlation between the histological changes and glucose intestinal absorption following a single dose of 5 fluorouracil. , 1970, Digestion.