Effect of large colon ischemia and reperfusion on concentrations of calprotectin and other clinicopathologic variables in jugular and colonic venous blood in horses

Astrid Grosche Island Whirl Equine Colic Research Laboratory, Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610.
Shands Transplant Center at University of Florida, University of Florida Health, Gainesville, FL 32608.

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Alison J. Morton Island Whirl Equine Colic Research Laboratory, Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610.

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A. Sarah Graham Island Whirl Equine Colic Research Laboratory, Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610.

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Maximilian M. R. Polyak Shands Transplant Center at University of Florida, University of Florida Health, Gainesville, FL 32608.

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David E. Freeman Island Whirl Equine Colic Research Laboratory, Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610.

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Abstract

Objective—To determine the effect of large colon ischemia and reperfusion on concentrations of the inflammatory neutrophilic protein calprotectin and other clinicopathologic variables in jugular and colonic venous blood in horses.

Animals—6 healthy horses.

Procedures—Horses were anesthetized, and ischemia was induced for 1 hour followed by 4 hours of reperfusion in a segment of the pelvic flexure of the large colon. Blood samples were obtained before anesthesia, before induction of ischemia, 1 hour after the start of ischemia, and 1, 2, and 4 hours after the start of reperfusion from jugular veins and veins of the segment of the large colon that underwent ischemia and reperfusion. A sandwich ELISA was developed for detection of equine calprotectin. Serum calprotectin concentrations and values of blood gas, hematologic, and biochemical analysis variables were determined.

Results—Large colon ischemia caused metabolic acidosis, a significant increase in lactate and potassium concentrations and creatine kinase activities, and a nonsignificant decrease in glucose concentrations in colonic venous blood samples. Values of these variables after reperfusion were similar to values before ischemia. Ischemia and reperfusion induced activation of an inflammatory response characterized by an increase in neutrophil cell turnover rate in jugular and colonic venous blood samples and calprotectin concentrations in colonic venous blood samples.

Conclusions and Clinical Relevance—Results of this study suggested that large colon ischemia and reperfusion caused local and systemic inflammation in horses. Serum calprotectin concentration may be useful as a marker of this inflammatory response.

Abstract

Objective—To determine the effect of large colon ischemia and reperfusion on concentrations of the inflammatory neutrophilic protein calprotectin and other clinicopathologic variables in jugular and colonic venous blood in horses.

Animals—6 healthy horses.

Procedures—Horses were anesthetized, and ischemia was induced for 1 hour followed by 4 hours of reperfusion in a segment of the pelvic flexure of the large colon. Blood samples were obtained before anesthesia, before induction of ischemia, 1 hour after the start of ischemia, and 1, 2, and 4 hours after the start of reperfusion from jugular veins and veins of the segment of the large colon that underwent ischemia and reperfusion. A sandwich ELISA was developed for detection of equine calprotectin. Serum calprotectin concentrations and values of blood gas, hematologic, and biochemical analysis variables were determined.

Results—Large colon ischemia caused metabolic acidosis, a significant increase in lactate and potassium concentrations and creatine kinase activities, and a nonsignificant decrease in glucose concentrations in colonic venous blood samples. Values of these variables after reperfusion were similar to values before ischemia. Ischemia and reperfusion induced activation of an inflammatory response characterized by an increase in neutrophil cell turnover rate in jugular and colonic venous blood samples and calprotectin concentrations in colonic venous blood samples.

Conclusions and Clinical Relevance—Results of this study suggested that large colon ischemia and reperfusion caused local and systemic inflammation in horses. Serum calprotectin concentration may be useful as a marker of this inflammatory response.

Contributor Notes

Supported by the American College of Veterinary Surgeons and the University of Florida College of Veterinary Medicine.

The authors thank Linda Green for technical assistance with development of the calprotectin ELISA.

Address correspondence to Dr. Grosche (agrosche@ufl.edu).
  • 1. Snyder JR. The pathophysiology of intestinal damage: effects of luminal distension and ischemia. Vet Clin North Am Equine Pract 1989; 5:247270.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 2. Gibson KT, Steel CM. Strangulating obstructions of the large colon in mature horses. Equine Vet Educ 1999; 11:234242.

  • 3. Moore RM, Muir WW, Granger DN. Mechanisms of gastrointestinal ischemia-reperfusion injury and potential therapeutic interventions: a review and its implications in the horse. J Vet Intern Med 1995; 9:115132.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 4. McAnulty JF, Stone WC, Darien BJ. The effects of ischemia and reperfusion on mucosal respiratory function, adenosine triphosphate, electrolyte, and water content in the ascending colon of ponies. Vet Surg 1997; 26:172181.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 5. Eltzschig HK, Eckle T. Ischemia and reperfusion—from mechanism to translation. Nature Med 2011; 17:13911401.

  • 6. Rowe EL, White NA. Reperfusion injury in the equine intestine. Clin Tech Equine Pract 2002; 1:148162.

  • 7. Fink MP, Delude RL. Epithelial barrier dysfunction: a unifying theme to explain the pathogenesis of multiple organ dysfunction at the cellular level. Crit Care Clin 2005; 21:177196.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 8. Vollmar B, Menger MD. Intestinal ischemia/reperfusion: micro-circulatory pathology and functional consequences. Langenbecks Arch Surg 2011; 396:13129.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 9. Gruys E, Toussaint MJM, Niewold TA, et al. Acute phase reaction and acute phase proteins. J Zhejiang Univ Sci 2005;6B:10451056.

  • 10. Cray C, Zaias J, Altman NH. Acute phase response in animals: a review. Comp Med 2009; 59:517526.

  • 11. Ihler CF, Venger JL, Skjerve E. Evaluation of clinical and laboratory variables as prognostic indicators in hospitalized gastrointestinal colic horses. Acta Vet Scand 2004; 45:109118.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 12. Saulez MN, Cebra CK, Tornquist SJ. The diagnostic and prognostic value of alkaline phosphatase activity in serum and peritoneal fluid from horses with acute colic. J Vet Intern Med 2004; 18:564567.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 13. Grosche A, Schrödl W, Schusser GF. Spezifische Parameter im Blut und Bauchpunktat zur Ermittlung des Schweregrades von intestinaler Ischämie bei Kolikpferden. Tierärztl Prax 2006; 34:387396.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 14. Johnston K, Holcombe SJ, Hauptman JG. Plasma lactate as a predictor of colonic viability and survival after 360° volvulus of the ascending colon in horses. Vet Surg 2007; 36:563567.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 15. Moore RM, Muir WW, Bertone AL, et al. Characterization of hemodynamic and metabolic alterations in the large colon of horses during low-flow ischemia and reperfusion. Am J Vet Res 1994; 55:14441453.

    • Search Google Scholar
    • Export Citation
  • 16. Kawcak CE, Baxter GM, Getzy DM, et al. Abnormalities in oxygenation, coagulation, and fibrinolysis in colonic blood of horses with experimentally induced strangulation obstruction. Am J Vet Res 1995; 56:16421650.

    • Search Google Scholar
    • Export Citation
  • 17. Moore RM, Couto CG, Muir WW, et al. Systemic and colonic venous hemostatic alterations in horses during low-flow ischemia and reperfusion of the large colon. Am J Vet Res 1995; 56:664670.

    • Search Google Scholar
    • Export Citation
  • 18. Moore RM, Muir WW, Rush BR. Systemic and colonic venous plasma biochemical alterations in horses during low-flow ischemia and reperfusion of the large colon. Can J Vet Res 1998; 62:1420.

    • Search Google Scholar
    • Export Citation
  • 19. Weiss DJ, Evanson OA. Evaluation of activated neutrophils in the blood of horses with colic. Am J Vet Res 2003; 64:13641368.

  • 20. D'Inca R, Dal Pont E, Di Leo V, et al. Calprotectin and lactoferrin in the assessment of intestinal inflammation and organic disease. Int J Colorectal Dis 2007; 22:429437.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 21. Grulke S, Franck T, Gangl M, et al. Myeloperoxidase assay in plasma and peritoneal fluid of horses with gastrointestinal disease. Can J Vet Res 2008; 72:3742.

    • Search Google Scholar
    • Export Citation
  • 22. De la Rebiere de Pouyade G, Riggs LM, Moore JN, et al. Equine neutrophil elastase in plasma, laminar tissue, and skin of horses administered black walnut heartwood extract. Vet Immunol Immunopathol 2010; 135:181187.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 23. Foell D, Frosch M, Sorg C, et al. Phagocyte-specific calcium-binding S100 proteins as clinical laboratory markers of inflammation. Clin Chim Acta 2004; 344:3751.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 24. Striz I, Trebichavský I. Calprotectin—a pleiotropic molecule in acute and chronic inflammation. Physiol Res 2004; 53:245253.

  • 25. Little D, Tomlinson JE, Blikslager AT. Post operative neutrophilic inflammation in equine small intestine after manipulation and ischaemia. Equine Vet J 2005; 37:329335.

    • Search Google Scholar
    • Export Citation
  • 26. Grosche A, Morton AJ, Polyak MMR, et al. Detection of calprotectin and its correlation to the accumulation of neutrophils within equine large colon during ischaemia and reperfusion. Equine Vet J 2008; 40:393399.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 27. Hsu K, Champaiboon C, Brian D. et al. Anti-infective protective properties of s100 calgranulin. Antiinflamm Antiallergy Agents Med Chem 2009; 8:290305.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 28. Voganatsi A, Panyutich A, Miyasaki KT, et al. Mechanism of extracellular release of human neutrophil calprotectin complex. J Leuko Biol 2001; 70:130134.

    • Search Google Scholar
    • Export Citation
  • 29. Fagerhol MK, Dale I, Andersson T. A radioimmunoassay for a granulocyte protein as a marker in studies on the turnover of such cells. Bull Eur Physiopathol Resp Suppl 1980; (16):273282.

    • Search Google Scholar
    • Export Citation
  • 30. Ivanov GE, Misuno NI, Ivanov VE. Enzyme-linked immunosorbent assay for human MRP-8/MRP-14 proteins and their quantitation in plasma of hematological patients. Immunol Lett 1996; 49:713.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 31. Heilmann RM, Suchodolski JS, Steiner JM. Development and analytic validation of a radioimmunoassay for the quantification of canine calprotectin in serum and feces from dogs. Am J Vet Res 2008; 69:845853.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 32. Graham AS, Grosche A, Morton AJ, et al. In vitro and in vivo responses of large colon mucosa to ischemia and reperfusion. Am J Vet Res 2011; 72:982989.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 33. Yousefi R, Ardestani SK, Saboury AA, et al. Investigation on the surface hydrophobicity and aggregation kinetics of human calprotectin in the presence of calcium. J Biochem Mol Biol 2005; 38:407413.

    • Search Google Scholar
    • Export Citation
  • 34. Medicineworld website. Calculators. Available at: www.medicineworld.org/calculators/bicarbonate/bicarbonate-base-excess-calculator.html. Accessed Mar 28, 2012.

    • Search Google Scholar
    • Export Citation
  • 35. Burnell JM, Villamil MF, Uyeno BT, et al. The effect in humans of extracellular pH change on the relationship between serum potassium concentration and intracellular potassium. J Clin Invest 1956; 35:935939.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 36. Hoskote SS, Joshi SR, Ghosh AK. Disorders of potassium homeostasis: pathophysiology and management. J Assoc Phys India 2008; 56:685693.

    • Search Google Scholar
    • Export Citation
  • 37. Fried MW, Murthy UK, Hassig SR, et al. Creatine kinase isoenzymes in the diagnosis of intestinal infarction. Dig Dis Sci 1991; 6:15891593.

    • Search Google Scholar
    • Export Citation
  • 38. Mukai M, Tamaki T, Noto T, et al. A new mechanism of serum creatine phosphokinase elevation in strangulated small bowel obstruction: an experimental rat model. J Internat Med Res 1995; 23:184190.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 39. Graeber GM, Cafferty PJ, Wolf RE, et al. An analysis of creatine phosphokinase in the mucosa and the muscularis of the gastrointestinal tract. J Surg Res 1984; 37:376382.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 40. Wallimann T, Hemmer T. Creatine kinase in non-muscle tissues and cells. Mol Cell Biochem 1994; 133/134:193220.

  • 41. Graeber GM, Wukich DK, Cafferty PJ, et al. Changes in peripheral serum creatine phosphokinase (CPK) and lactic dehydrogenase (LDH) in acute experimental colonic infarction. Ann Surg 1981; 194:708715.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 42. Boyd JW. The mechanisms relating to increases in plasma enzymes and isoenzymes in diseases of animals. Vet Clin Pathol 1983; 12:924.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 43. Grosche A, Morton AJ, Graham AS, et al. Ultrastructural changes in the equine colonic mucosa after ischemia and reperfusion. Equine Vet J 2011; 43:815.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 44. Grosche A, Morton AJ, Graham AS, et al. Mucosal injury and inflammatory cells in response to brief ischemia and reperfusion in the equine large colon. Equine Vet J 2011; 43:1625.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 45. Kaminski KA, Bonda TA, Korecki J, et al. Oxidative stress and neutrophil activation—the two keystones of ischemia/reperfusion injury. Internat J Cardiol 2002; 86:4159.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 46. Kalia N, Brown NJ, Wood RFM, et al. Effects of intestinal ischemia-reperfusion injury on rat peripheral blood neutrophil activation. Dig Dis Sci 2003; 48:16771684.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 47. Souza DG, Cara DC, Cassali GD, et al. Effects of the PAF receptor antagonist UK74505 on local and remote reperfusion injuries following ischaemia of the superior mesenteric artery in the rat. Br J Pharmacol 2000; 131:18001808.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 48. Granger DN, Korthuis RJ. Physiologic mechanisms of postischemic tissue injury. Ann Rev Physiol 1995; 57:311332.

  • 49. Nathan C. Neutrophils and immunity: challenges and opportunities. Nature Rev Immunol 2006; 6:173182.

  • 50. Mantovani A, Cassatella MA, Costantini C, et al. Neutrophils in the activation and regulation of innate and adaptive immunity. Nature Rev Immunol 2011; 11:519531.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 51. Guan Y, Worrell RT, Pritts TA, et al. Intestinal ischemia-reperfusion injury: reversible and irreversible damage imaged in vivo. Am J Physiol Gastrointest Liver Physiol 2009; 297:G187G196.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 52. Kono H, Rock KL. How dying cells alert the immune system to danger. Nature Rev Immunol 2008; 8:279289.

  • 53. Rock KL, Kono H. The inflammtory response to cell death. Ann Rev Pathol 2008; 3:99126.

  • 54. Johne B, Fagerhol MK, Lyberg T, et al. Functional and clinical aspects of the myelomonocyte protein calprotectin. Mol Pathol 1997; 50:113123.

  • 55. Heilmann RM, Jergens AE, Ackermann MR, et al. Serum calprotectin concentrations in dogs with idiopathic inflammatory bowel disease. Am J Vet Res 2012; 73:19001907.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 56. Fagerhol MK, Dale I, Andersson T. Release and quantification of a leucocyte derived protein (L1). Scand J Haematol 1980; 24:393398.

    • Search Google Scholar
    • Export Citation
  • 57. McConnico RS, Weinstock D, Poston ME, et al. Myeloperoxidase activity of the large intestine in an equine model of acute colitis. Am J Vet Res 1999; 60:807813.

    • Search Google Scholar
    • Export Citation
  • 58. Deby-Dupont G, Deby C, Lamy M. Neutrophil myeloperoxidase revisited: it's role in health and disease. Intensivmedizin 1999; 36:500513.

  • 59. Rötting AK, Freeman DE, Constable P, et al. Mucosal distribution of eosinophilic granulocytes within the gastrointestinal tract of horses. Am J Vet Res 2008; 69:874879.

    • Crossref
    • Search Google Scholar
    • Export Citation

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