Effects of continuous or intermittent lipopolysaccharide administration for 48 hours on the systemic inflammatory response in horses

Elizabeth M. Tadros Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, TN 37996.

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Nicholas Frank Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, TN 37996.

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Abstract

Objective—To determine whether the method of lipopolysaccharide (LPS) administration (intermittent vs continuous) affects the magnitude and duration of the systemic inflammatory response in horses and whether prolonged (48 hours) endotoxemia induces laminitis.

Animals—12 healthy adult horses (10 mares and 2 geldings).

Procedures—Horses were randomly assigned to receive LPS (total dose, 80 μg; n = 4) or saline (0.9% NaCl) solution (80 mL/h; 4) via constant rate infusion or 8 bolus IV injections of LPS (10 μg, q 6 h;4) during a 48-hour period. Physical examinations were performed every 4 hours, inflammatory cytokine gene expression was determined for blood samples obtained every 8 hours, and IV glucose tolerance tests were performed.

Results—All LPS-treated horses had signs of depression and mild colic; those signs abated as the study progressed. Administration of LPS increased expression of interleukin-1β, interleukin-6, and interleukin-8, but results were not significantly different between LPS treatment groups. Cytokine expression was significantly higher on the first day versus the second day of LPS treatment. Interleukin-1β expression was positively correlated with rectal temperature and expression of other cytokines. Glucose and insulin dynamics for both LPS groups combined did not differ significantly from those of the saline solution group. Signs of laminitis were not detected in any of the horses.

Conclusions and Clinical Relevance—Horses developed LPS tolerance within approximately 24 hours after administration was started, and the method of LPS administration did not affect the magnitude or duration of systemic inflammation. Laminitis was not induced in horses.

Abstract

Objective—To determine whether the method of lipopolysaccharide (LPS) administration (intermittent vs continuous) affects the magnitude and duration of the systemic inflammatory response in horses and whether prolonged (48 hours) endotoxemia induces laminitis.

Animals—12 healthy adult horses (10 mares and 2 geldings).

Procedures—Horses were randomly assigned to receive LPS (total dose, 80 μg; n = 4) or saline (0.9% NaCl) solution (80 mL/h; 4) via constant rate infusion or 8 bolus IV injections of LPS (10 μg, q 6 h;4) during a 48-hour period. Physical examinations were performed every 4 hours, inflammatory cytokine gene expression was determined for blood samples obtained every 8 hours, and IV glucose tolerance tests were performed.

Results—All LPS-treated horses had signs of depression and mild colic; those signs abated as the study progressed. Administration of LPS increased expression of interleukin-1β, interleukin-6, and interleukin-8, but results were not significantly different between LPS treatment groups. Cytokine expression was significantly higher on the first day versus the second day of LPS treatment. Interleukin-1β expression was positively correlated with rectal temperature and expression of other cytokines. Glucose and insulin dynamics for both LPS groups combined did not differ significantly from those of the saline solution group. Signs of laminitis were not detected in any of the horses.

Conclusions and Clinical Relevance—Horses developed LPS tolerance within approximately 24 hours after administration was started, and the method of LPS administration did not affect the magnitude or duration of systemic inflammation. Laminitis was not induced in horses.

Contributor Notes

Dr. Tadros' present address is Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.

Dr. Frank's present address is Department of Clinical Sciences, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA 01536.

Supported by the University of Tennessee College of Veterinary Medicine Center of Excellence, the Charles and Julie Wharton Fellowship, and the Wells Graduate Student Fellowship.

Presented as a poster presentation at the American College of Veterinary Internal Medicine Forum, Anaheim, Calif, June 2010.

The authors thank Dr. David Horohov for technical assistance with the real-time PCR assay techniques and for providing primer sequences and Dr. Arnold Saxton for assistance with the statistical analysis.

Address correspondence to Dr. Frank (nicholas.frank@tufts.edu).
  • 1.

    Parsons CSOrsini JAKrafty R, et al. Risk factors for development of acute laminitis in horses during hospitalization: 73 cases (1997–2004). J Am Vet Med Assoc 2007; 230:885889.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 2.

    Senior JMProudman CJLeuwer M, et al. Plasma endotoxin in horses presented to an equine referral hospital: correlation to selected clinical parameters and outcomes. Equine Vet J 2011; 43:585591.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 3.

    Werners AHBull SFink-Gremmels J. Endotoxaemia: a review with implications for the horse. Equine Vet J 2005; 37:371383.

  • 4.

    Belknap JK. Black walnut extract: an inflammatory model. Vet Clin North Am Equine Pract 2010; 26:95101.

  • 5.

    Belknap JKMoore JNCrouser EC. Sepsis–from human organ failure to laminar failure. Vet Immunol Immunopathol 2009; 129:155157.

  • 6.

    Belknap JKGiguere SPettigrew A, et al. Lamellar pro-inflammatory cytokine expression patterns in laminitis at the developmental stage and at the onset of lameness: innate vs. adaptive immune response. Equine Vet J 2007; 39:4247.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 7.

    Loftus JPBlack SJPettigrew A, et al. Early laminar events involving endothelial activation in horses with black walnut–induced laminitis. Am J Vet Res 2007; 68:12051211.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 8.

    Budak MTOrsini JAPollitt CC, et al. Gene expression in the lamellar dermis-epidermis during the developmental phase of carbohydrate overload-induced laminitis in the horse. Vet Immunol Immunopathol 2009; 131:8696.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 9.

    Noschka EVandenplas MLHurley DJ, et al. Temporal aspects of laminar gene expression during the developmental stages of equine laminitis. Vet Immunol Immunopathol 2009; 129:242253.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 10.

    Leise BSFaleiros RRWatts M, et al. Laminar inflammatory gene expression in the carbohydrate overload model of equine laminitis. Equine Vet J 2011; 43:5461.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 11.

    Hurley DJParks RJReber AJ, et al. Dynamic changes in circulating leukocytes during the induction of equine laminitis with black walnut extract. Vet Immunol Immunopathol 2006; 110:195206.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 12.

    Stewart AJPettigrew ACochran AM, et al. Indices of inflammation in the lung and liver in the early stages of the black walnut extract model of equine laminitis. Vet Immunol Immunopathol 2009; 129:254260.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 13.

    de la Rebiere de Pouyade GRiggs LMMoore 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
  • 14.

    Nieto JEMacDonald MHBraim AEP, et al. Effect of lipopolysaccharide infusion on gene expression of inflammatory cytokines in normal horses in vivo. Equine Vet J 2009; 41:717719.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 15.

    MacKay RJMerritt AMZertuche JM, et al. Tumor necrosis factor activity in the circulation of horses given endotoxin. Am J Vet Res 1991; 52:533538.

    • Search Google Scholar
    • Export Citation
  • 16.

    MacKay RJLester GD. Induction of the acute-phase cytokine, hepatocyte-stimulating factor/interleukin 6, in the circulation of horses treated with endotoxin. Am J Vet Res 1992; 53:12851289.

    • Search Google Scholar
    • Export Citation
  • 17.

    Turek JJTempleton CBBottoms GD, et al. Flunixin meglumine attenuation of endotoxin-induced damage to the cardiopulmonary vascular endothelium of the pony. Am J Vet Res 1985; 46:591596.

    • Search Google Scholar
    • Export Citation
  • 18.

    Ward DSFessler JFBottoms GD, et al. Equine endotoxemia: cardiovascular, eicosanoid, hematologic, blood chemical, and plasma enzyme alterations. Am J Vet Res 1987; 48:11501156.

    • Search Google Scholar
    • Export Citation
  • 19.

    Duncan SGMeyers KMReed SM, et al. Alterations in coagulation and hemograms of horses given endotoxins for 24 hours via hepatic portal infusions. Am J Vet Res 1985; 46:12871293.

    • Search Google Scholar
    • Export Citation
  • 20.

    Buras JAHolzmann BSitkovsky M. Animal models of sepsis: setting the stage. Nat Rev Drug Discovery 2005; 4:854865.

  • 21.

    Murphy TJPaterson HMKriynovich S, et al. Linking the “two-hit” response following injury to enhanced TLR4 reactivity. J Leukoc Biol 2005; 77:1623.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 22.

    Steinberg JHalter JSchiller H, et al. The development of acute respiratory distress syndrome after gut ischemia/reperfusion injury followed by fecal peritonitis in pigs: a clinically relevant model. Shock 2005; 23:129137.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 23.

    Marik PERaghavan M. Stress-hyperglycemia, insulin and immunomodulation in sepsis. Intensive Care Med 2004; 30:748756.

  • 24.

    Toth FFrank NElliott SB, et al. Effects of an intravenous endotoxin challenge on glucose and insulin dynamics in horses. Am J Vet Res 2008; 69:8288.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 25.

    Toth FFrank NChameroy KA, et al. Effects of endotoxaemia and carbohydrate overload on glucose and insulin dynamics and the development of laminitis in horses. Equine Vet J 2009; 41:852858.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 26.

    Morris DDMoore JN. Antibody titres to core lipopolysaccharides in horses with gastrointestinal disorders which cause colic. Equine Vet J Suppl 1989; 7:2932.

    • Search Google Scholar
    • Export Citation
  • 27.

    Taylor DHood DMWagner IP. Short-term effect of therapeutic shoeing on severity of lameness in horses with chronic laminitis. Am J Vet Res 2002; 63:16291633.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 28.

    Livak KJSchmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25:402408.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 29.

    Freestone JFWolfsheimer KJKamerling SG, et al. Exercise induced hormonal and metabolic changes in Thoroughbred horses: effects of conditioning and acepromazine. Equine Vet J 1991; 23:219223.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 30.

    Piccione GCaola GRefinetti R. The circadian rhythm of body temperature of the horse. Biol Rhythm Res 2002; 33:113119.

  • 31.

    Allen GKCampbell-Beggs CRobinson JA, et al. Induction of early-phase endotoxin tolerance in horses. Equine Vet J 1996; 28:269274.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 32.

    Scumpia POMoldawer LL. Biology of interleukin-10 and its regulatory roles in sepsis syndromes. Crit Care Med 2005; 33:S468S471.

  • 33.

    Lin ECalvano SELowry SF. Inflammatory cytokines and cell response in surgery. Surgery 2000; 127:117126.

  • 34.

    Boontham PChandran PRowlands B, et al. Surgical sepsis: dysregulation of immune function and therapeutic implications. Surgeon 2003; 1:187206.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 35.

    Rainen LOelmueller UJurgensen S, et al. Stabilization of mRNA expression in whole blood samples. Clin Chem 2002; 48:18831890.

  • 36.

    Perneger TV. What's wrong with Bonferroni adjustments. Br Med J 1998; 316:12361238.

  • 37.

    Fan HCook JA. Molecular mechanisms of endotoxin tolerance. J Endotoxin Res 2004; 10:7184.

  • 38.

    Biswas SKLopez-Collazo E. Endotoxin tolerance: new mechanisms, molecules and clinical significance. Trends Immunol 2009; 30:475487.

  • 39.

    Cavaillon JMAdrie CFitting C, et al. Reprogramming of circulatory cells in sepsis and SIRS. J Endotoxin Res 2005; 11:311320.

  • 40.

    Cavaillon JMAnnane D. Compartmentalization of the inflammatory response in sepsis and SIRS. J Endotoxin Res 2006; 12:151170.

  • 41.

    Lang JDHickman-Davis JM. One-hit, two-hit … is there really any benefit? Clin Exp Immunol 2005; 141:211214.

  • 42.

    Moine PShenkar RKaneko D, et al. Systemic blood loss affects NF-kappa B regulatory mechanisms in the lungs. Am J Physiol 1997; 273:L185L192.

    • Search Google Scholar
    • Export Citation
  • 43.

    Smith PDSuffredini AFAllen JB, et al. Endotoxin administration to humans primes alveolar macrophages for increased production of inflammatory mediators. J Clin Immunol 1994; 14:141148.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 44.

    Parker LCJones ECPrince LR, et al. Endotoxin tolerance induces selective alterations in neutrophil function. J Leukoc Biol 2005; 78:13011305.

  • 45.

    Barton MHParviainen ANorton N. Polymyxin B protects horses against induced endotoxaemia in vivo. Equine Vet J 2004; 36:397401.

  • 46.

    Kelmer GDoherty TJElliott S, et al. Evaluation of dimethyl sulphoxide effects on initial response to endotoxin in the horse. Equine Vet J 2008; 40:358363.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 47.

    Poulin Braim AEMacDonald MHBruss ML, et al. Effects of intravenous administration of pirfenidone on horses with experimentally induced endotoxemia. Am J Vet Res 2009; 70:10311042.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 48.

    Garrett LABrown RPoxton IR. A comparative study of the intestinal microbiota of healthy horses and those suffering from equine grass sickness. Vet Microbiol 2002; 87:8188.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 49.

    Elliott JBailey SR. Gastrointestinal derived factors are potential triggers for the development of acute equine laminitis. J Nutr 2006; 136:2103S2107S.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 50.

    Crawford CSepulveda MFElliott J, et al. Dietary fructan carbohydrate increases amine production in the equine large intestine: implications for pasture-associated laminitis. J Anim Sci 2007; 85:29492958.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 51.

    Bailey SRAdair HSReinemeyer CR, et al. Plasma concentrations of endotoxin and platelet activation in the developmental stage of oligofructose-induced laminitis. Vet Immunol Immunopathol 2009; 129:167173.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 52.

    Zerpa HVega FVasquez J, et al. Effect of acute sublethal endotoxaemia on in vitro digital vascular reactivity in horses. J Vet Med A Physiol Pathol Clin Med 2005; 52:6773.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 53.

    Bailey SRElliott J. The role of prostanoids and nitric oxide in endotoxin-induced hyporesponsiveness of equine digital blood vessels. Equine Vet J 1999; 31:212218.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 54.

    Riggs LMKrunkosky TMNoschka E, et al. Comparison of characteristics and enzymatic products of leukocytes in the skin and laminar tissues of horses administered black walnut heartwood extract or lipopolysaccharide. Am J Vet Res 2009; 70:13831390.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 55.

    Hanly BKStokes AMBell AM, et al. Use of serial laminar tissue collection via biopsy in conscious healthy horses. Am J Vet Res 2009; 70:697702.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 56.

    Toth FFrank NGeor RJ, et al. Effects of pretreatment with dexamethasone or levothyroxine sodium on endotoxin-induced alterations in glucose and insulin dynamics in horses. Am J Vet Res 2010; 71:6068.

    • Crossref
    • Search Google Scholar
    • Export Citation

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