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Effects of indomethacin on the rat small intestinal mucosa: immunohistochemical and biochemical studies using anti-mucin monoclonal antibodies

  • Original Article—Alimentary Tract
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Abstract

Background

The luminal surface of the gastrointestinal tract is covered by a viscoelastic gel layer that acts as a protective barrier against the intraluminal environment. Because the situation of the small intestine has not been elucidated to the same degree as other sections, in this study, we investigated the effects of indomethacin on the rat small intestinal mucosa.

Methods

Male Wistar rats were given indomethacin 10 mg/kg s-c and sacrificed 1, 3, 7, or 14 days later. The small intestine was opened along the anti-mesenteric side, and examined macroscopically. Total mucin content in the small intestinal epithelium was measured and immunoreactivity was examined using anti-mucin monoclonal antibodies HCM31 and PGM34.

Results

Indomethacin caused punched out and linear ulcers located mostly along the mesenteric margin of the distal jejunum with sparing of the ileum. Histological examination showed sialomucin recognized by HCM31 increased on day 3 especially in the regenerating epithelium around the ulcer edge. Furthermore, the surface mucous gel layer displayed a multilaminated pattern, consisting of non-sulfated sialomucin-rich layers and sulfated mucin-rich layers, where both mucins had the common core protein, MUC2. Biochemical measurements also showed the total mucin content of the jejunum increased transiently and HCM31-positive mucin increased approximately 4 times greater than baseline on day 3, but no marked changes were observed in the ileum, with few ulcers observed.

Conclusions

Indomethacin administration causes quantitative and qualitative change in jejunal mucin. In particular, sialomucin plays an important role in regenerating epithelium during the healing process following indomethacin-induced mucosal damage.

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References

  1. Walker WA. Development of the intestinal mucosal barrier. J Pediatr Gastroenterol Nutr. 2002;34(Suppl 1):S33–9.

    Article  PubMed  CAS  Google Scholar 

  2. Corfield AP, Myerscough N, Longman R, Sylvester P, Arul S, Pignatelli M. Mucins and mucosal protection in the gastrointestinal tract: new prospects for mucins in the pathology of gastrointestinal disease. Gut. 2000;47:589–94.

    Article  PubMed  CAS  Google Scholar 

  3. Takeuchi K, Miyazawa T, Tanaka A, Kato S, Kunikata T. Pathogenic importance of intestinal hypermotility in NSAID-induced small intestinal damage in rats. Digestion. 2002;66:30–41.

    Article  PubMed  CAS  Google Scholar 

  4. Tanaka A, Matsumoto M, Hayashi Y, Takeuchi K. Functional mechanism underlying cyclooxygenase-2 expression in rat small intestine following administration of indomethacin: relation to intestinal hypermotility. J Gastroenterol Hepatol. 2005;20:38–45.

    Article  PubMed  CAS  Google Scholar 

  5. Satoh H, Guth PH, Grossman MI. Role of food in gastrointestinal ulceration produced by indomethacin in the rat. Gastroenterology. 1982;83:210–5.

    PubMed  CAS  Google Scholar 

  6. Evans SM, Whittle BJ. Role of bacteria and inducible nitric oxide synthase activity in the systemic inflammatory microvascular response provoked by indomethacin in the rat. Eur J Pharmacol. 2003;461:63–71.

    Article  PubMed  CAS  Google Scholar 

  7. Hagiwara M, Kataoka K, Arimochi H, Kuwahara T, Ohnishi Y. Role of unbalanced growth of gram-negative bacteria in ileal ulcer formation in rats treated with a nonsteroidal anti-inflammatory drug. J Med Invest. 2004;51:43–51.

    Article  PubMed  Google Scholar 

  8. Reuter BK, Davies NM, Wallace JL. Nonsteroidal anti-inflammatory drug enteropathy in rats: role of permeability, bacteria, and enterohepatic circulation. Gastroenterology. 1997;112:109–17.

    Article  PubMed  CAS  Google Scholar 

  9. Hollander F. The two-component mucous barrier; its activity in protecting the gastroduodenal mucosa against peptic ulceration. AMA Arch Intern Med. 1954;93:107–20.

    PubMed  CAS  Google Scholar 

  10. Menguy R. Gastric mucus and the gastric mucous barrier. A review. Am J Surg. 1969;117:806–12.

    Article  PubMed  CAS  Google Scholar 

  11. Allen A. Structure of gastrointestinal mucus glycoproteins and the viscous and gel-forming properties of mucus. Br Med Bull. 1978;34:28–33.

    PubMed  CAS  Google Scholar 

  12. Keshav S, Lawson L, Chung LP, Stein M, Perry VH, Gordon S. Tumor necrosis factor mRNA localized to Paneth cells of normal murine intestinal epithelium by in situ hybridization. J Exp Med. 1990;171:327–32.

    Article  PubMed  CAS  Google Scholar 

  13. Molmenti EP, Perlmutter DH, Rubin DC. Cell-specific expression of alpha 1-antitrypsin in human intestinal epithelium. J Clin Invest. 1993;92:2022–34.

    Article  PubMed  CAS  Google Scholar 

  14. Peeters T, Vantrappen G. The Paneth cell: a source of intestinal lysozyme. Gut. 1975;16:553–8.

    Article  PubMed  CAS  Google Scholar 

  15. Meyer-Hoffert U, Hornef MW, Henriques-Normark B, Axelsson LG, Midtvedt T, Putsep K, Andersson M. Secreted enteric antimicrobial activity localizes to the mucus surface layer. Gut. 2008;57:764–71.

    Article  PubMed  CAS  Google Scholar 

  16. Azuumi Y, Ohara S, Ishihara K, Okabe H, Hotta K. Correlation of quantitative changes of gastric mucosal glycoproteins with aspirin-induced gastric damage in rats. Gut. 1980;21:533–6.

    Article  PubMed  CAS  Google Scholar 

  17. Ikezawa T, Ichikawa T, Adachi K, Sugano S, Ojima T, Nakamura Y, et al. Analysis of mucin composition in gastric juices of chronic rheumatic patients with upper gastrointestinal damage. Biomed Res. 2005;26:147–51.

    Article  PubMed  CAS  Google Scholar 

  18. Kojima Y, Ishihara K, Komuro Y, Saigenji K, Hotta K. Effects of the muscarinic receptor agonist carbachol and/or antagonist pirenzepine on gastric mucus secretion in rats. Scand J Gastroenterol. 1993;28:647–51.

    Article  PubMed  CAS  Google Scholar 

  19. Ikezawa T, Goso Y, Ichikawa T, Hayashida H, Kurihara M, Okayasu I, et al. Appearance of specific mucins recognized by monoclonal antibodies in rat gastric mucosa healing from HCl-induced gastric mucosal damage. J Gastroenterol. 2004;39:113–9.

    Article  PubMed  CAS  Google Scholar 

  20. Ishihara K, Kurihara M, Goso Y, Ota H, Katsuyama T, Hotta K. Establishment of monoclonal antibodies against carbohydrate moiety of gastric mucins distributed in the different sites and layers of rat gastric mucosa. Glycoconj J. 1996;13:857–64.

    Article  PubMed  CAS  Google Scholar 

  21. Ikezawa T, Goso Y, Ichikawa T, Hayashida H, Nakamura T, Kurihara M, et al. Immunohistochemical localization in rat gastrointestinal tract of a sialomucin species recognized by HCM31, a new anti-mucin monoclonal antibody. Biomed Res. 2002;23:63–8.

    CAS  Google Scholar 

  22. Tsubokawa D, Goso Y, Sawaguchi A, Kurihara M, Ichikawa T, Sato N, et al. A monoclonal antibody, PGM34, against 6-sulfated blood-group H type 2 antigen, on the carbohydrate moiety of mucin. FEBS J. 2007;274:1833–48.

    Article  PubMed  CAS  Google Scholar 

  23. Anthony A, Dhillon AP, Thrasivoulou C, Pounder RE, Wakefield AJ. Pre-ulcerative villous contraction and microvascular occlusion induced by indomethacin in the rat jejunum: a detailed morphological study. Aliment Pharmacol Ther. 1995;9:605–13.

    PubMed  CAS  Google Scholar 

  24. Wax J, Clinger WA, Varner P, Bass P, Winder CV. Relationship of the enterohepatic cycle to ulcerogenesis in the rat small bowel with flufenamic acid. Gastroenterology. 1970;58:772–80.

    PubMed  CAS  Google Scholar 

  25. Anthony A, Pounder RE, Dhillon AP, Wakefield AJ. Vascular anatomy defines sites of indomethacin induced jejunal ulceration along the mesenteric margin. Gut. 1997;41:763–70.

    Article  PubMed  CAS  Google Scholar 

  26. Saegusa Y, Ichikawa T, Iwai T, Goso Y, Okayasu I, Ikezawa T, Shikama N, Saigenji K, Ishihara K. Changes in the mucus barrier of the rat during 5-fluorouracil-induced gastrointestinal mucositis. Scand J Gastroenterol. 2008;43:59–65.

    Article  PubMed  CAS  Google Scholar 

  27. Kato S, Tanaka A, Kunikata T, Umeda M, Takeuchi K. Protective effect of lafutidine against indomethacin-induced intestinal ulceration in rats: relation to capsaicin-sensitive sensory neurons. Digestion. 2000;61:39–46.

    Article  PubMed  CAS  Google Scholar 

  28. Hatazawa R, Ohno R, Tanigami M, Tanaka A, Takeuchi K. Roles of endogenous prostaglandins and cyclooxygenase isozymes in healing of indomethacin-induced small intestinal lesions in rats. J Pharmacol Exp Ther. 2006;318:691–9.

    Article  PubMed  CAS  Google Scholar 

  29. Azuumi Y, Ishihara K, Ohara S, Okabe H, Hotta K. Efficacy of anti-ulcer drugs on the recovery of gastric mucosal glycoproteins with aspirin-induced gastric damage in rat. Gastroenterol Jpn. 1981;16:331–4.

    PubMed  CAS  Google Scholar 

  30. Kojima Y, Ishihara K, Ohara S, Saigenji K, Hotta K. Effects of the M1 muscarinic receptor antagonist pirenzepine on gastric mucus glycoprotein in rats with or without ethanol-induced gastric damage. Scand J Gastroenterol. 1992;27:764–8.

    Article  PubMed  CAS  Google Scholar 

  31. Yusuf S, Nok AJ, Ameh DA, Adelaiye AB, Balogun EO. Correlation of gastric mucosal damage with sialic acid profile in rats: effect of hydrochloric acid, pepsin and hypertonic saline. Cell Biochem Funct. 2005;23:339–45.

    Article  PubMed  CAS  Google Scholar 

  32. Ho SB, Takamura K, Anway R, Shekels LL, Toribara NW, Ota H. The adherent gastric mucous layer is composed of alternating layers of MUC5AC and MUC6 mucin proteins. Dig Dis Sci. 2004;49:1598–606.

    Article  PubMed  CAS  Google Scholar 

  33. Matsuo K, Ota H, Akamatsu T, Sugiyama A, Katsuyama T. Histochemistry of the surface mucous gel layer of the human colon. Gut. 1997;40:782–9.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We express our sincere thanks to S. Sugawara, M. Hashimura, and Y. Numata for technical assistance. Part of this work was supported by Grants from the Shin-Caterpillar Mitsubishi Co., Grants from the SRL Co., and Grants from the Integrative Research Project of the Graduate School of Medical Sciences, Kitasato University.

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Authors and Affiliations

  1. Department of Gastroenterology, Kitasato University School of Medicine, Sagamihara, Japan

    Tomohisa Iwai, Tomoaki Ikezawa, Yoichi Saegusa & Katsunori Saigenji

  2. Department of Biochemistry, Kitasato University School of Medicine, Sagamihara, Japan

    Takafumi Ichikawa

  3. Department of Regulation Biochemistry, Kitasato University Graduate School of Medical Sciences, 1-15-1, Kitasato, Sagamihara, Kanagawa, 228-8555, Japan

    Yukinobu Goso & Kazuhiko Ishihara

  4. Department of Pathology, Kitasato University School of Medicine, 1-15-1 Kitasato, Sagamihara, 228-8555, Japan

    Isao Okayasu

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  1. Tomohisa Iwai
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  2. Takafumi Ichikawa
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  3. Yukinobu Goso
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  4. Tomoaki Ikezawa
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  5. Yoichi Saegusa
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  8. Kazuhiko Ishihara
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Correspondence to Kazuhiko Ishihara.

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Iwai, T., Ichikawa, T., Goso, Y. et al. Effects of indomethacin on the rat small intestinal mucosa: immunohistochemical and biochemical studies using anti-mucin monoclonal antibodies. J Gastroenterol 44, 277–284 (2009). https://doi.org/10.1007/s00535-009-0007-0

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  • DOI: https://doi.org/10.1007/s00535-009-0007-0

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