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. 1999 Oct;45(4):516–522. doi: 10.1136/gut.45.4.516

Metabolism of oral trefoil factor 2 (TFF2) and the effect of oral and parenteral TFF2 on gastric and duodenal ulcer healing in the rat

S Poulsen 1, J Thulesen 1, L Christensen 1, E Nexo 1, L Thim 1
PMCID: PMC1727673  PMID: 10486358

Abstract

BACKGROUND—Trefoil factors (TFFs) are peptides produced by mucus-secreting cells in the gastrointestinal tract. A functional association between these peptides and mucus, leading to stabilisation of the viscoelastic gel overlying the epithelia, has been suggested. Both oral and parenteral administration of the peptides increase the resistance of the gastric mucosa.
AIM—To study the effect in rats of oral and parenteral porcine trefoil factor 2 (pTFF2) on the healing of gastric and duodenal ulcerations and to clarify the distribution and metabolism of orally administered pTFF2 in the gastrointestinal tract.
METHODS—Gastric ulcers were induced in female Sprague-Dawley rats by indomethacin and duodenal ulcers by mercaptamine. The rats were treated for up to seven days with oral or subcutaneous pTFF2. Ulcer size after treatment was assessed by stereomicroscopy after whole mount staining with periodic acid-Schiff stain. 125I-labelled pTFF2 was given orally to rats, and tissues were investigated by gamma counting of samples and by autoradiography of paraffin embedded sections.
RESULTS—pTFF2 accelerated gastric ulcer healing after both oral and subcutaneous administration. Duodenal ulcers were aggravated by both treatments. After oral administration of 125I-pTFF2, intact peptide was recovered from the superficial part of the mucus layer in the stomach; it passed through the small intestine but was degraded in the caecum. Only a minor part of the labelled pTFF2 entered the colon and was excreted in the faeces. Most of the label was excreted in the urine.
CONCLUSIONS—Oral as well as parenteral pTFF2 accelerates the healing of gastric ulceration and aggravates duodenal ulcers. Oral pTFF2 binds to the mucus layer of the stomach and the small intestine but does not reach the colonic mucosa.


Keywords: trefoil factors; spasmolytic polypeptide; ulcer healing; gastric ulcer; duodenal ulcer; rat

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Figure 1  .

Figure 1  

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Rat stomach with indomethacin induced ulcers after whole mount staining with periodic acid-Schiff. The specimen has been photographed with a Wild photomacroscope. The borderline of the rumen is in the upper part of the photograph. The gastro-oesophageal junction is indicated by a large arrow and the ulcers are seen as well defined white areas (small arrows). Original magnification × 4. 

Figure 2  .

Figure 2  

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(A) The effect of oral treatment with porcine trefoil factor 2 (pTFF2) on the healing of gastric ulceration expressed as the total area of ulceration in mm2 in the stomach after seven days of treatment (mean (SD), n = 7). *p<0.05 v controls. (B) The effect of oral treatment for one and three days on the healing of duodenal ulceration expressed as ulcerated area in mm2 (mean (SD), n = 8). *p<0.05 v controls.

Figure 3  .

Figure 3  

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(A) Effect of subcutaneous treatment with porcine trefoil factor 2 (pTFF2) and omeprazole for three and seven days on the healing of gastric ulceration expressed as the total area of ulceration in mm2 in the stomach (mean (SD), n = 7). *p<0.05 v controls. (B) Effect of subcutaneous treatment for three days on the healing of duodenal ulceration expressed as ulcerated area in mm2 (mean (SD), n = 7). *p<0.05 v controls.

Figure 4  .

Figure 4  

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Percentage of total 125I-pTFF2 radioactivity given orally to each rat present in the various parts of the gastrointestinal tract after 0.5, 2, 4, 12, and 24 hours. The small intestine is divided into 10 cm segments. Results are mean (SD) (n = 4).

Figure 5  .

Figure 5  

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Autoradiograph showing grains in the superficial part of the mucus layer (arrows) in the stomach, four hours after oral administration of 125I-labelled porcine trefoil factor 2. Original magnification × 550. 

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Babyatsky M. W., deBeaumont M., Thim L., Podolsky D. K. Oral trefoil peptides protect against ethanol- and indomethacin-induced gastric injury in rats. Gastroenterology. 1996 Feb;110(2):489–497. doi: 10.1053/gast.1996.v110.pm8566596. [DOI] [PubMed] [Google Scholar]
  2. Chinery R., Cox H. M. Immunoprecipitation and characterization of a binding protein specific for the peptide, intestinal trefoil factor. Peptides. 1995;16(4):749–755. doi: 10.1016/0196-9781(95)00045-l. [DOI] [PubMed] [Google Scholar]
  3. Dignass A., Lynch-Devaney K., Kindon H., Thim L., Podolsky D. K. Trefoil peptides promote epithelial migration through a transforming growth factor beta-independent pathway. J Clin Invest. 1994 Jul;94(1):376–383. doi: 10.1172/JCI117332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hanby A. M., Poulsom R., Singh S., Elia G., Jeffery R. E., Wright N. A. Spasmolytic polypeptide is a major antral peptide: distribution of the trefoil peptides human spasmolytic polypeptide and pS2 in the stomach. Gastroenterology. 1993 Oct;105(4):1110–1116. doi: 10.1016/0016-5085(93)90956-d. [DOI] [PubMed] [Google Scholar]
  5. Hauser F., Poulsom R., Chinery R., Rogers L. A., Hanby A. M., Wright N. A., Hoffmann W. hP1.B, a human P-domain peptide homologous with rat intestinal trefoil factor, is expressed also in the ulcer-associated cell lineage and the uterus. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):6961–6965. doi: 10.1073/pnas.90.15.6961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hoffmann W., Hauser F. The P-domain or trefoil motif: a role in renewal and pathology of mucous epithelia? Trends Biochem Sci. 1993 Jul;18(7):239–243. doi: 10.1016/0968-0004(93)90170-r. [DOI] [PubMed] [Google Scholar]
  7. Jeffrey G. P., Oates P. S., Wang T. C., Babyatsky M. W., Brand S. J. Spasmolytic polypeptide: a trefoil peptide secreted by rat gastric mucous cells. Gastroenterology. 1994 Feb;106(2):336–345. doi: 10.1016/0016-5085(94)90590-8. [DOI] [PubMed] [Google Scholar]
  8. Jørgensen K. H., Thim L., Jacobsen H. E. Pancreatic spasmolytic polypeptide (PSP): I. Preparation and initial chemical characterization of a new polypeptide from porcine pancreas. Regul Pept. 1982 Mar;3(3-4):207–219. doi: 10.1016/0167-0115(82)90126-4. [DOI] [PubMed] [Google Scholar]
  9. Mashimo H., Wu D. C., Podolsky D. K., Fishman M. C. Impaired defense of intestinal mucosa in mice lacking intestinal trefoil factor. Science. 1996 Oct 11;274(5285):262–265. doi: 10.1126/science.274.5285.262. [DOI] [PubMed] [Google Scholar]
  10. McKenzie C., Marchbank T., Playford R. J., Otto W., Thim L., Parsons M. E. Pancreatic spasmolytic polypeptide protects the gastric mucosa but does not inhibit acid secretion or motility. Am J Physiol. 1997 Jul;273(1 Pt 1):G112–G117. doi: 10.1152/ajpgi.1997.273.1.G112. [DOI] [PubMed] [Google Scholar]
  11. Playford R. J., Marchbank T., Chinery R., Evison R., Pignatelli M., Boulton R. A., Thim L., Hanby A. M. Human spasmolytic polypeptide is a cytoprotective agent that stimulates cell migration. Gastroenterology. 1995 Jan;108(1):108–116. doi: 10.1016/0016-5085(95)90014-4. [DOI] [PubMed] [Google Scholar]
  12. Playford R. J., Marchbank T., Goodlad R. A., Chinery R. A., Poulsom R., Hanby A. M. Transgenic mice that overexpress the human trefoil peptide pS2 have an increased resistance to intestinal damage. Proc Natl Acad Sci U S A. 1996 Mar 5;93(5):2137–2142. doi: 10.1073/pnas.93.5.2137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Podolsky D. K., Lynch-Devaney K., Stow J. L., Oates P., Murgue B., DeBeaumont M., Sands B. E., Mahida Y. R. Identification of human intestinal trefoil factor. Goblet cell-specific expression of a peptide targeted for apical secretion. J Biol Chem. 1993 Mar 25;268(9):6694–6702. [PubMed] [Google Scholar]
  14. Poulsen S. S., Olsen P. S., Kirkegaard P. Healing of cysteamine-induced duodenal ulcers in the rat. Dig Dis Sci. 1985 Feb;30(2):161–167. doi: 10.1007/BF01308204. [DOI] [PubMed] [Google Scholar]
  15. Poulsen S. S., Thulesen J., Nexø E., Thim L. Distribution and metabolism of intravenously administered trefoil factor 2/porcine spasmolytic polypeptide in the rat. Gut. 1998 Aug;43(2):240–247. doi: 10.1136/gut.43.2.240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rasmussen T. N., Raaberg L., Poulsen S. S., Thim L., Holst J. J. Immunohistochemical localization of pancreatic spasmolytic polypeptide (PSP) in the pig. Histochemistry. 1992 Sep;98(2):113–119. doi: 10.1007/BF00717002. [DOI] [PubMed] [Google Scholar]
  17. Rio M. C., Bellocq J. P., Daniel J. Y., Tomasetto C., Lathe R., Chenard M. P., Batzenschlager A., Chambon P. Breast cancer-associated pS2 protein: synthesis and secretion by normal stomach mucosa. Science. 1988 Aug 5;241(4866):705–708. doi: 10.1126/science.3041593. [DOI] [PubMed] [Google Scholar]
  18. Rio M. C., Chenard M. P., Wolf C., Marcellin L., Tomasetto C., Lathe R., Bellocq J. P., Chambon P. Induction of pS2 and hSP genes as markers of mucosal ulceration of the digestive tract. Gastroenterology. 1991 Feb;100(2):375–379. doi: 10.1016/0016-5085(91)90205-y. [DOI] [PubMed] [Google Scholar]
  19. Sands B. E., Podolsky D. K. The trefoil peptide family. Annu Rev Physiol. 1996;58:253–273. doi: 10.1146/annurev.ph.58.030196.001345. [DOI] [PubMed] [Google Scholar]
  20. Selye H., Szabo S. Experimental model for production of perforating duodenal ulcers by cysteamine in the rat. Nature. 1973 Aug 17;244(5416):458–459. doi: 10.1038/244458a0. [DOI] [PubMed] [Google Scholar]
  21. Suemori S., Lynch-Devaney K., Podolsky D. K. Identification and characterization of rat intestinal trefoil factor: tissue- and cell-specific member of the trefoil protein family. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11017–11021. doi: 10.1073/pnas.88.24.11017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Takeuchi K., Okada M., Ebara S., Osano H. Increased microvascular permeability and lesion formation during gastric hypermotility caused by indomethacin and 2-deoxy-D-glucose in the rat. J Clin Gastroenterol. 1990;12 (Suppl 1):S76–S84. doi: 10.1097/00004836-199001001-00014. [DOI] [PubMed] [Google Scholar]
  23. Tan X. D., Hsueh W., Chang H., Wei K. R., Gonzalez-Crussi F. Characterization of a putative receptor for intestinal trefoil factor in rat small intestine: identification by in situ binding and ligand blotting. Biochem Biophys Res Commun. 1997 Aug 28;237(3):673–677. doi: 10.1006/bbrc.1997.7144. [DOI] [PubMed] [Google Scholar]
  24. Thim L. A new family of growth factor-like peptides. 'Trefoil' disulphide loop structures as a common feature in breast cancer associated peptide (pS2), pancreatic spasmolytic polypeptide (PSP), and frog skin peptides (spasmolysins). FEBS Lett. 1989 Jun 19;250(1):85–90. doi: 10.1016/0014-5793(89)80690-8. [DOI] [PubMed] [Google Scholar]
  25. Thim L., Jørgensen K. H., Jørgensen K. D. Pancreatic spasmolytic polypeptide (PSP): II. Radioimmunological determination of PSP in porcine tissues, plasma and pancreatic juice. Regul Pept. 1982 Mar;3(3-4):221–230. doi: 10.1016/0167-0115(82)90127-6. [DOI] [PubMed] [Google Scholar]
  26. Thim L. Trefoil peptides: from structure to function. Cell Mol Life Sci. 1997 Dec;53(11-12):888–903. doi: 10.1007/s000180050108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Williams R., Stamp G. W., Gilbert C., Pignatelli M., Lalani E. N. pS2 transfection of murine adenocarcinoma cell line 410.4 enhances dispersed growth pattern in a 3-D collagen gel. J Cell Sci. 1996 Jan;109(Pt 1):63–71. doi: 10.1242/jcs.109.1.63. [DOI] [PubMed] [Google Scholar]
  28. Wright N. A., Hoffmann W., Otto W. R., Rio M. C., Thim L. Rolling in the clover: trefoil factor family (TFF)-domain peptides, cell migration and cancer. FEBS Lett. 1997 May 19;408(2):121–123. doi: 10.1016/s0014-5793(97)00424-9. [DOI] [PubMed] [Google Scholar]
  29. Wright N. A., Poulsom R., Stamp G. W., Hall P. A., Jeffery R. E., Longcroft J. M., Rio M. C., Tomasetto C., Chambon P. Epidermal growth factor (EGF/URO) induces expression of regulatory peptides in damaged human gastrointestinal tissues. J Pathol. 1990 Dec;162(4):279–284. doi: 10.1002/path.1711620402. [DOI] [PubMed] [Google Scholar]
  30. Wright N. A., Poulsom R., Stamp G., Van Noorden S., Sarraf C., Elia G., Ahnen D., Jeffery R., Longcroft J., Pike C. Trefoil peptide gene expression in gastrointestinal epithelial cells in inflammatory bowel disease. Gastroenterology. 1993 Jan;104(1):12–20. doi: 10.1016/0016-5085(93)90830-6. [DOI] [PubMed] [Google Scholar]

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