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Home / Equine Research Bank / Equine Vet J / Influence of digital hypothermia on lamellar events related ...
Equine veterinary journal2019; 52(3); 441-448; doi: 10.1111/evj.13184

Influence of digital hypothermia on lamellar events related to IL-6/gp130 signalling in equine sepsis-related laminitis.

Abstract: Interleukin-6 (IL-6) is consistently increased in the digital lamellae in different studies of sepsis-related laminitis (SRL). IL-6 signalling through the gp130 receptor activates similar signalling (i.e. mTORC1-related signalling) previously reported to be activated in models of endocrinopathic laminitis. Objective: To assess the activation state of signalling proteins downstream of IL-6/gp130 receptor complex activation in an experimental model of SRL. Methods: Randomised experimental study. Methods: Lamellar phospho-(P) protein concentrations downstream of the IL-6/gp130 receptors were assessed in the oligofructose (OF) model of SRL. Fifteen Standardbred horses were administered water (CON, n = 8) or oligofructose (OF, n = 7) via a nasogastric tube. At 12 h post-OF/water administration, one randomly assigned forelimb was exposed to continuous digital hypothermia (CDH) by placement in ice water (ICE, maintained at <7°C); the other forelimb was maintained at ambient temperature (AMB). Lamellar tissue samples were collected after 24 h of CDH from both ICE and AMB forelimbs and immediately snap-frozen. Lamellar proteins of interest were assessed by immunoblotting and immunofluorescence. Results: Immunoblotting revealed increase (P<0.05) in the phosphorylation states of Akt (Ser 473), RPS6 (Ser235/236), RPS6 (Ser240/244), STAT3 (Ser727) and STAT3 (Tyr705) in lamellar tissue from OF-treated animals (AMB OF vs. AMB CON limbs); CDH resulted in decreased (P<0.05) lamellar concentrations of phosphorylated Akt, p70S6K, RPS6 (235/236), RPS6 (240/244) and STAT3 (S727) in OF-treated animals (AMB OF vs. ICE OF). Immunofluorescence showed that activated/phosphorylated forms of RPS6 and STAT3 were primarily localised to lamellar epithelial cells. Conclusions: The nature, sequence and timing of sub-cellular events in this experimental model may differ from those that accompany naturally occurring sepsis. Conclusions: There were increased lamellar concentrations of activated signalling proteins downstream of the IL-6/Gp130 receptor complex in OF-treated horses; CDH inhibited this activation for the majority of the proteins assessed. These results demonstrate similar lamellar signalling (e.g. mTORC1-related signalling) and, therefore, possible therapeutic targets occurring in sepsis-related laminitis as previously reported in models of endocrinopathic laminitis.
© 2019 EVJ Ltd.
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Publication Date: 2019-10-24 PubMed ID: 31509270DOI: 10.1111/evj.13184Google Scholar: Lookup
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  • Journal Article

Summary

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The research investigates the response of proteins associated with the IL-6/Gp130 receptor complex during sepsis-related laminitis in horses. It uncovers that sepsis-related laminitis provokes increased levels of activated signaling proteins downstream of the IL-6/Gp130 receptor complex, but this activation can be restricted by continuous digital hypothermia.

Research Objective and Methods

The aim of this study was to understand the activity state of signaling proteins downstream of the IL-6/gp130 signaling complex in a lab test of sepsis-related laminitis (SRL) in horses.

  • Two types of treatments were used in this study: water (control group) and oligofructose (for inducing SRL).
  • The researchers used 15 Standardbred horses, randomly distributing them between the two treatments.
  • One leg from each horse was kept at regular room temperature, while the other leg was treated with continuous digital hypothermia by being submerged in ice water.
  • After 24 hours of continuous digital hypothermia, lamellar tissue samples were collected from both legs and analyzed using immunoblotting and immunofluorescence methods.

Research Findings

The data generated from immunoblotting and immunofluorescence indicate that:

  • sepsis-related laminitis prompts an increase in activated signaling proteins, specifically phosphorylated Akt, RPS6, and STAT3 in the lamellar tissue of oligofructose-treated horses.
  • continuous digital hypothermia reduces the concentration of these phosphorylated proteins.
  • the proteins are mainly localized in the lamellar epithelial cells.

Conclusions

The changes in the activities of sub-cellular events in this controlled experiment may differ from naturally occurring sepsis. However, main findings concluded that:

  • therapeutic targets for sepsis-related laminitis may be shared with endocrinopathic laminitis due to similar lamellar signaling processes seen in IL-6/Gp130 receptor complex.
  • these activated signaling proteins could be potential therapeutic targets.
  • continuous digital hypothermia may be an effective method of inhibiting their activation.

Cite This Article

APA
Dern K, Burns TA, Watts MR, van Eps AW, Belknap JK. (2019). Influence of digital hypothermia on lamellar events related to IL-6/gp130 signalling in equine sepsis-related laminitis. Equine Vet J, 52(3), 441-448. https://doi.org/10.1111/evj.13184

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 52
Issue: 3
Pages: 441-448

Researcher Affiliations

Dern, K
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Ohio State University, Columbus, Ohio, USA.
Burns, T A
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Ohio State University, Columbus, Ohio, USA.
Watts, M R
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Ohio State University, Columbus, Ohio, USA.
van Eps, A W
  • Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Belknap, J K
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Ohio State University, Columbus, Ohio, USA.

MeSH Terms

  • Animals
  • Cytokine Receptor gp130
  • Foot Diseases / veterinary
  • Hoof and Claw
  • Horse Diseases
  • Horses
  • Hypothermia / veterinary
  • Inflammation / veterinary
  • Interleukin-6
  • Sepsis / veterinary

Grant Funding

  • Grayson-Jockey Club Research Foundation

References

This article includes 39 references
  1. Iskander KN, Osuchowski MF, Stearns-Kurosawa DJ, Kurosawa S, Stepien D, Valentine C, Remick DG. Sepsis: multiple abnormalities, heterogeneous responses, and evolving understanding. Physiol. Rev. 93, 1247-1288.
  2. van der Poll T, van de Veerdonk FL, Scicluna BP, Netea MG. The immunopathology of sepsis and potential therapeutic targets. Nat. Rev. Immunol. 17, 407-420.
  3. Marshall JC. Why have clinical trials in sepsis failed?. Trends Mol. Med. 20, 195-203.
  4. Crouser ED. Mitochondrial dysfunction in septic shock and multiple organ dysfunction syndrome. Mitochondrion 4, 729-741.
  5. Belknap JK, Moore JN, Crouser EC. Sepsis-From human organ failure to laminar failure. Vet. Immunol. Immunopathol. 129, 155-157.
  6. Stewart AJ, Pettigrew A, Cochran AM, Belknap JK. Indices of inflammation in the lung and liver in the early stages of the black walnut extract model of equine laminitis. Vet. Immunol. Immunopathol. 129, 254-260.
  7. Black SJ. Extracellular matrix, leukocyte migration and laminitis. Vet. Immunol. Immunopathol. 129, 161-163.
  8. Belknap JK, Giguere S, Pettigrew A, Cochran AM, Van Eps AW, Pollitt CC. 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. 39, 42-47.
  9. van Eps AW, Walters LJ, Baldwin GI, McGarry M, Pollitt CM. Distal limb cryotherapy for the prevention of acute laminitis. Clin. Tech. Equine Pract. 3, 64-70.
  10. van Eps AW, Pollitt CC, Underwood C, Medina-Torres CE, Goodwin WA, Belknap JK. Continuous digital hypothermia initiated after the onset of lameness prevents lamellar failure in the oligofructose laminitis model. Equine Vet. J. 46, 625-630.
  11. Kullman A, Holcombe SJ, Hurcombe SD, Roessner HA, Hauptman JG, Geor RJ, Belknap JK. Prophylactic digital cryotherapy is associated with decreased incidence of laminitis in horses diagnosed with colitis. Equine Vet. J. 46, 554-559.
  12. van Eps AW, Leise BS, Watts M, Pollitt CC, Belknap JK. Digital hypothermia inhibits early lamellar inflammatory signalling in the oligofructose laminitis model. Equine Vet. J. 44, 230-237.
  13. Godman JD, Burns TA, Kelly CS, Watts MR, Leise BS, Schroeder EL, van Eps AW, Belknap JK. The effect of hypothermia on influx of leukocytes in the digital lamellae of horses with oligofructose-induced laminitis. Vet. Immunol. Immunopathol. 178, 22-28.
  14. Dern K, Watts M, Werle B, van Eps A, Pollitt C, Belknap J. Effect of delayed digital hypothermia on lamellar inflammatory signaling in the oligofructose laminitis model. J. Vet. Intern. Med. 31, 575-581.
  15. Thiem S, Pierce TP, Palmieri M, Putoczki TL, Buchert M, Preaudet A, Farid RO, Love C, Catimel B, Lei Z, Rozen S, Gopalakrishnan V, Schaper F, Hallek M, Boussioutas A, Tan P, Jarnicki A, Ernst M. mTORC1 inhibition restricts inflammation-associated gastrointestinal tumorigenesis in mice. J. Clin. Invest. 123, 767-781.
  16. Bharti R, Dey G, Mandal M. Cancer development, chemoresistance, epithelial to mesenchymal transition and stem cells: a snapshot of IL-6 mediated involvement. Cancer Lett. 375, 51-61.
  17. de Laat MA, Patterson-Kane JC, Pollitt CC, Sillence MN, McGowan CM. Histological and morphometric lesions in the pre-clinical, developmental phase of insulin-induced laminitis in Standardbred horses. Vet. J. 195, 305-312.
  18. French KR, Pollitt CC. Equine laminitis: loss of hemidesmosomes in hoof secondary epidermal lamellae correlates to dose in an oligofructose induction model: an ultrastructural study. Equine Vet. J. 36, 230-235.
  19. Nourian AR, Asplin KE, McGowan CM, Sillence MN, Pollitt CC. Equine laminitis: ultrastructural lesions detected in ponies following hyperinsulinaemia. Equine Vet. J. 41, 671-677.
  20. Nourian AR, Baldwin GI, van Eps AW, Pollitt CC. Equine laminitis: ultrastructural lesions detected 24-30 hours after induction with oligofructose. Equine Vet. J. 39, 360-364.
  21. Pollitt CC. Basement membrane pathology: a feature of acute equine laminitis. Equine Vet. J. 28, 38-46.
  22. Savagner P. Epithelial-mesenchymal transitions: from cell plasticity to concept elasticity. Curr. Top. Dev. Biol. 112, 273-300.
  23. Sun BO, Fang Y, Li Z, Chen Z, Xiang J. Role of cellular cytoskeleton in epithelial-mesenchymal transition process during cancer progression. Biomed. Rep. 3, 603-610.
  24. Bollrath J, Phesse TJ, von Burstin VA, Putoczki T, Bennecke M, Bateman T, Nebelsiek T, Lundgren-May T, Canli O, Schwitalia S, Matthews V, Schmid RM, Kirchner T, Arkan MC, Ernst M, Greten FR. gp130-mediated Stat3 activation in enterocytes regulates cell survival and cell-cycle progression during colitis-associated tumorigenesis. Cancer Cell 15, 91-102.
  25. Bromberg J, Wang TC. Inflammation and cancer: IL-6 and STAT3 complete the link. Cancer Cell 15, 79-80.
  26. Lane HE, Burns TA, Hegedus OC, Watts MR, Weber PS, Woltman KA, Geor RJ, McCutcheon LJ, Eades SC, Mathes LE, Belknap JK. Lamellar events related to insulin-like growth factor-1 receptor signalling in two models relevant to endocrinopathic laminitis. Equine Vet. J. 49, 643-654.
  27. Dern K, van Eps A, Wittum T, Watts M, Pollitt C, Belknap J. Effect of continuous digital hypothermia on lamellar inflammatory signaling when applied at a clinically-relevant timepoint in the oligofructose laminitis model. J. Vet. Intern. Med. 32, 450-458.
  28. van Eps AW, Pollitt CC. Equine laminitis induced with oligofructose. Equine Vet. J. 38, 203-238.
  29. Ip CK, Cheung AN, Ngan HY, Wong AS. p70 S6 kinase in the control of actin cytoskeleton dynamics and directed migration of ovarian cancer cells. Oncogene 30, 2420-2432.
  30. Mok KW, Chen H, Lee WM, Cheng CY. rpS6 regulates blood-testis barrier dynamics through Arp3-mediated actin microfilament organization in rat sertoli cells. An in vitro study. Endocrinology 156, 1900-1913.
  31. Liu L, Luo Y, Chen L, Shen T, Xu B, Chen W, Zhou H, Han X, Huang S. Rapamycin inhibits cytoskeleton reorganization and cell motility by suppressing RhoA expression and activity. J. Biol. Chem. 285, 38362-38373.
  32. Liu L, Chen L, Chung J, Huang S. Rapamycin inhibits F-actin reorganization and phosphorylation of focal adhesion proteins. Oncogene 27, 4998-5010.
  33. Liu L, Parent CA. Review series: TOR kinase complexes and cell migration. J. Cell. Biol. 194, 815-824.
  34. Chen X, Cheng H, Pan T, Liu Y, Su Y, Ren C, Huang D, Zha X, Liang C. mTOR regulate EMT through RhoA and Rac1 pathway in prostate cancer. Mol. Carcinog. 54, 1086-1095.
  35. Yang GS, Zhou XY, An XF, Liu XJ, Zhang YJ, Yu D. mTOR is involved in stroke-induced seizures and the anti-seizure effect of mild hypothermia. Mol. Med. Rep. 17, 5821-5829.
  36. Kim JH, Yoon MS, Chen J. Signal transducer and activator of transcription 3 (STAT3) mediates amino acid inhibition of insulin signaling through serine 727 phosphorylation. J. Biol. Chem. 284, 35425-35432.
  37. Mao X, Cho MJT, Ellebrecht CT, Mukherjee EM, Payne AS. Stat3 regulates desmoglein 3 transcription in epithelial keratinocytes. JCI Insight. 2.
    doi: 10.1172/jci.insight.92253google scholar: lookup
  38. Garbers C, Aparicio-Siegmund S, Rose-John S. The IL-6/gp130/STAT3 signaling axis: recent advances towards specific inhibition. Curr. Opin. Immunol. 34, 75-82.
  39. Baran P, Hansen S, Waetzig GH, Akbarzadeh M, Lamertz L, Huber HJ, Ahmadian MR, Moll JM, Scheller J. The balance of interleukin (IL)-6, IL-6*soluble IL-6 receptor (sIL-6R), and IL-6*sIL-6R*sgp130 complexes allows simultaneous classic and trans-signaling. J. Biol. Chem. 293, 6762-6775.

Citations

This article has been cited 3 times.
  1. Burns TA, Watts MR, Belknap JK, van Eps AW. Digital lamellar inflammatory signaling in an experimental model of equine preferential weight bearing.. J Vet Intern Med 2023 Mar;37(2):681-688.
    doi: 10.1111/jvim.16662pubmed: 36840365google scholar: lookup
  2. Guo HH, Jing XY, Chen H, Xu HX, Zhu BM. STAT3 but Not STAT5 Contributes to the Protective Effect of Electroacupuncture Against Myocardial Ischemia/Reperfusion Injury in Mice.. Front Med (Lausanne) 2021;8:649654.
    doi: 10.3389/fmed.2021.649654pubmed: 34307396google scholar: lookup
  3. Stokes SM, Burns TA, Watts MR, Bertin FR, Stefanovski D, Medina-Torres CE, Belknap JK, van Eps AW. Effect of digital hypothermia on lamellar inflammatory signaling in the euglycemic hyperinsulinemic clamp laminitis model.. J Vet Intern Med 2020 Jul;34(4):1606-1613.
    doi: 10.1111/jvim.15835pubmed: 32583504google scholar: lookup
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