• 1. Ohta H, Sunden Y, Yokoyama N, et al. Expression of apical junction complex proteins in duodenal mucosa of dogs with inflammatory bowel disease. Am J Vet Res 2014; 75: 746751.

    • Search Google Scholar
    • Export Citation
  • 2. Rioux KP, Madsen KL, Fedorak RN. The role of enteric microflora in inflammatory bowel disease: human and animal studies with probiotics and prebiotics. Gastroenterol Clin North Am 2005; 34: 465482.

    • Search Google Scholar
    • Export Citation
  • 3. Packey CD, Sartor RB. Commensal bacteria, traditional and opportunistic pathogens, dysbiosis and bacterial killing in inflammatory bowel diseases. Curr Opin Infect Dis 2009; 22: 292301.

    • Search Google Scholar
    • Export Citation
  • 4. Allenspach K, House A, Smith K, et al. Evaluation of mucosal bacteria and histopathology, clinical disease activity and expression of Toll-like receptors in German Shepherd Dogs with chronic enteropathies. Vet Microbiol 2010; 146: 326335.

    • Search Google Scholar
    • Export Citation
  • 5. Suchodolski JS, Xenoulis PG, Paddock CG, et al. Molecular analysis of the bacterial microbiota in duodenal biopsies from dogs with idiopathic inflammatory bowel disease. Vet Microbiol 2010; 142: 394400.

    • Search Google Scholar
    • Export Citation
  • 6. Burgener IA, König A, Allenspach K, et al. Upregulation of toll-like receptors in chronic enteropathies in dogs. J Vet Intern Med 2008; 22: 553560.

    • Search Google Scholar
    • Export Citation
  • 7. Kathrani A, House A, Catchpole B, et al. Polymorphisms in the TLR4 and TLR5 gene are significantly associated with inflammatory bowel disease in German Shepherd Dogs. PLoS One 2010;5:e15740.

    • Search Google Scholar
    • Export Citation
  • 8. Goldberg RF, Austen WG Jr, Zhang X, et al. Intestinal alkaline phosphatase is a gut mucosal defense factor maintained by enteral nutrition. Proc Natl Acad Sci U S A 2008; 105: 35513556.

    • Search Google Scholar
    • Export Citation
  • 9. Tuin A, Poelstra K, de Jager-Krikken A, et al. Role of alkaline phosphatase in colitis in man and rats. Gut 2009; 58: 379387.

  • 10. Sisley AC, Desai TR, Hynes KL, et al. Decrease in mucosal alkaline phosphatase: a potential marker of intestinal reperfusion injury. J Lab Clin Med 1999; 133: 335341.

    • Search Google Scholar
    • Export Citation
  • 11. Bol-Schoenmakers M, Fiechter D, Raaben W, et al. Intestinal alkaline phosphatase contributes to the reduction of severe intestinal epithelial damage. Eur J Pharmacol 2010; 633: 7177.

    • Search Google Scholar
    • Export Citation
  • 12. Narisawa S, Huang L, Iwasaki A, et al. Accelerated fat absorption in intestinal alkaline phosphatase knockout mice. Mol Cell Biol 2003; 23: 75257530.

    • Search Google Scholar
    • Export Citation
  • 13. Akiba Y, Mizumori M, Guth PH, et al. Duodenal brush border intestinal alkaline phosphatase activity affects bicarbonate secretion in rats. Am J Physiol Gastrointest Liver Physiol 2007;293:G1223–G1233.

    • Search Google Scholar
    • Export Citation
  • 14. Mizumori M, Ham M, Guth PH, et al. Intestinal alkaline phosphatase regulates protective surface microclimate pH in rat duodenum. J Physiol 2009; 587: 36513663.

    • Search Google Scholar
    • Export Citation
  • 15. Lallès JP. Intestinal alkaline phosphatase: multiple biological roles in maintenance of intestinal homeostasis and modulation by diet. Nutr Rev 2010; 68: 323332.

    • Search Google Scholar
    • Export Citation
  • 16. Molnár K, Vannay A, Szebeni B, et al. Intestinal alkaline phosphatase in the colonic mucosa of children with inflammatory bowel disease. World J Gastroenterol 2012; 18: 32543259.

    • Search Google Scholar
    • Export Citation
  • 17. Day MJ, Bilzer T, Mansell J, et al. Histopathological standards for the diagnosis of gastrointestinal inflammation in endoscopic biopsy samples from the dog and cat: a report from the World Small Animal Veterinary Association Gastrointestinal Standardization Group. J Comp Pathol 2008; 138 (suppl 1): S1S43.

    • Search Google Scholar
    • Export Citation
  • 18. Allenspach K, Wieland B, Gröne A, et al. Chronic enteropathies in dogs: evaluation of risk factors for negative outcome. J Vet Intern Med 2007; 21: 700708.

    • Search Google Scholar
    • Export Citation
  • 19. Novikoff AB. Enzyme localizations with Wachstein-Meisel procedures: real or artifact. J Histochem Cytochem 1967; 15: 353354.

  • 20. Peters IR, Peeters D, Helps CR, et al. Development and application of multiple internal reference (housekeeper) gene assays for accurate normalisation of canine gene expression studies. Vet Immunol Immunopathol 2007; 117: 5566.

    • Search Google Scholar
    • Export Citation
  • 21. Engle MJ, Mahmood A, Alpers DH. Two rat intestinal alkaline phosphatase isoforms with different carboxyl-terminal peptides are both membrane-bound by a glycan phosphatidylinositol linkage. J Biol Chem 1995; 270: 1193511940.

    • Search Google Scholar
    • Export Citation
  • 22. Nakano T, Inoue I, Alpers DH, et al. Role of lysophosphatidylcholine in brush-border intestinal alkaline phosphatase release and restoration. Am J Physiol Gastrointest Liver Physiol 2009;297:G207–G214.

    • Search Google Scholar
    • Export Citation
  • 23. Poelstra K, Bakker WW, Klok P, et al. Dephosphorylation of endotoxin by alkaline phosphatase in vivo. Am J Pathol 1997; 151: 11631169.

    • Search Google Scholar
    • Export Citation
  • 24. Bentala H, Verweij WR, Huizinga-Van der Vlag A, et al. Removal of phosphate from lipid A as a strategy to detoxify lipopolysaccharide. Shock 2002; 18: 561566.

    • Search Google Scholar
    • Export Citation
  • 25. Beumer C, Wulferink M, Raaben W, et al. Calf intestinal alkaline phosphatase, a novel therapeutic drug for lipopolysaccharide (LPS)-mediated diseases, attenuates LPS toxicity in mice and piglets. J Pharmacol Exp Ther 2003; 307: 737744.

    • Search Google Scholar
    • Export Citation
  • 26. Poelstra K, Bakker WW, Klok PA, et al. A physiologic function for alkaline phosphatase: endotoxin detoxification. Lab Invest 1997; 76: 319327.

    • Search Google Scholar
    • Export Citation
  • 27. Bates JM, Akerlund J, Mittge E, et al. Intestinal alkaline phosphatase detoxifies lipopolysaccharide and prevents inflammation in zebrafish in response to the gut microbiota. Cell Host Microbe 2007; 2: 371382.

    • Search Google Scholar
    • Export Citation
  • 28. Vaishnava S, Hooper LV. Alkaline phosphatase: keeping the peace at the gut epithelial surface. Cell Host Microbe 2007; 2: 365367.

    • Search Google Scholar
    • Export Citation
  • 29. Xenoulis PG, Palculict B, Allenspach K, et al. Molecular-phylogenetic characterization of microbial communities imbalances in the small intestine of dogs with inflammatory bowel disease. FEMS Microbiol Ecol 2008; 66: 579589.

    • Search Google Scholar
    • Export Citation
  • 30. Malo MS, Biswas S, Abedrapo MA, et al. The pro-inflammatory cytokines, IL-1beta and TNF-alpha, inhibit intestinal alkaline phosphatase gene expression. DNA Cell Biol 2006; 25: 684695.

    • Search Google Scholar
    • Export Citation
  • 31. German AJ, Helps CR, Hall EJ, et al. Cytokine mRNA expression in mucosal biopsies from German Shepherd Dogs with small intestinal enteropathies. Dig Dis Sci 2000; 45: 717.

    • Search Google Scholar
    • Export Citation
  • 32. Peters IR, Helps CR, Calvert EL, et al. Cytokine mRNA quantification in duodenal mucosa from dogs with chronic enteropathies by real-time reverse transcriptase polymerase chain reaction. J Vet Intern Med 2005; 19: 644653.

    • Search Google Scholar
    • Export Citation
  • 33. Jergens AE, Sonea IM, O'Connor AM, et al. Intestinal cytokine mRNA expression in canine inflammatory bowel disease: a meta-analysis with critical appraisal. Comp Med 2009; 59: 153162.

    • Search Google Scholar
    • Export Citation
  • 34. Schmitz S, Garden OA, Werling D, et al. Gene expression of selected signature cytokines of T cell subsets in duodenal tissues of dogs with and without inflammatory bowel disease. Vet Immunol Immunopathol 2012; 146: 8791.

    • Search Google Scholar
    • Export Citation
  • 35. Wiedmeyer CE, Solter PE, Hoffmann WE. Alkaline phosphatase expression in tissues from glucocorticoid-treated dogs. Am J Vet Res 2002; 63: 10831088.

    • Search Google Scholar
    • Export Citation

Advertisement

Comparison of the expression, activity, and fecal concentration of intestinal alkaline phosphatase between healthy dogs and dogs with chronic enteropathy

View More View Less
  • 1 Laboratory of Veterinary Internal Medicine, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
  • | 2 Laboratory of Veterinary Internal Medicine, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
  • | 3 Laboratory of Veterinary Internal Medicine, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
  • | 4 Laboratory of Veterinary Internal Medicine, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
  • | 5 Laboratory of Veterinary Internal Medicine, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
  • | 6 Laboratory of Veterinary Internal Medicine, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.

Abstract

OBJECTIVE To compare expression, activity, and fecal concentration of intestinal alkaline phosphatase (IAP) between healthy dogs and dogs with chronic enteropathy (CE).

ANIMALS 9 healthy university-owned Beagles and 109 healthy client-owned dogs (controls) and 28 dogs with CE (cases).

PROCEDURES Cases were defined as dogs with persistent (> 3 weeks) gastrointestinal signs that failed to respond to antimicrobials and anti-inflammatory doses of prednisolone or dietary trials, did not have mechanical gastrointestinal abnormalities as determined by abdominal radiography and ultrasonography, and had a diagnosis of lymphoplasmacytic enteritis or eosinophilic gastroenteritis on histologic examination of biopsy specimens. Duodenal and colonic mucosa biopsy specimens were obtained from the 9 university-owned Beagles and all cases for histologic examination and determination of IAP expression (by real-time quantitative PCR assay) and activity (by enzyme histochemical analysis). Fecal samples were obtained from all dogs for determination of fecal IAP concentration by a quantitative enzyme reaction assay.

RESULTS For dogs evaluated, IAP expression and activity were localized at the luminal side of epithelial cells in the mucosa and intestinal crypts, although both were greater in the duodenum than in the colon. Active IAP was detected in the feces of all dogs. Intestinal alkaline phosphatase expression and activity were lower for cases than for controls, and fecal IAP concentration for dogs with moderate and severe CE was lower than that for dogs with mild CE.

CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that dogs with CE had impaired IAP expression and activity. Additional research is necessary to elucidate the role of IAP in the pathogenesis of CE.

Contributor Notes

Address correspondence to Dr. Ide (k-ide@cc.tuat.ac.jp).

Dr. Kato's present address is Ogata Animal Hospital, 1-14-23 Unomori, Minami-ku, Sagamihara, Kanagawa 252-0301, Japan. Dr. Sawa's present address is Mizuhodai Animal Hospital, 1-21-5 Nishi-Mizuhodai, Fujimi, Saitama 354-0018, Japan.