Cytokine mRNA expressions after racing at a high altitude and at sea level in horses with exercise-induced pulmonary hemorrhage

Montague N. Saulez Section of Equine Medicine, Department of Companion Animal Clinical Studies, Faculty of Veterinary Science, University of Pretoria, Pretoria 0110, South Africa.

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Jacques Godfroid Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Pretoria 0110, South Africa.

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Anamarie Bosman Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Pretoria 0110, South Africa.

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Jackie L. Stiltner Gluck Equine Research Center, Department of Veterinary Science, College of Agriculture, University of Kentucky, Lexington, KY 40546.

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Cormac C. Breathnach Gluck Equine Research Center, Department of Veterinary Science, College of Agriculture, University of Kentucky, Lexington, KY 40546.

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David W. Horohov Gluck Equine Research Center, Department of Veterinary Science, College of Agriculture, University of Kentucky, Lexington, KY 40546.

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Abstract

Objective—To determine concentrations of cytokine mRNA in horses with exercise-induced pulmonary hemorrhage (EIPH) after racing.

Animals—97 Thoroughbreds.

Procedures—Following tracheobronchoscopy, the severity of EIPH was graded (scale of 0 to 4), and venous blood samples were collected from 10 horses in each grade. After RNA isolation and cDNA synthesis, real-time PCR assay was conducted to detect cytokinespecific mRNA for interleukin (IL)-1, IL-6, and IL-10; interferon (INF)-γ; and tumor necrosis factor (TNF)-α.

Results—Neither location nor grade of EIPH affected the expression of IL-1 and INF-γ. There was significantly greater overall expression of IL-6 mRNA at sea level, with significantly more IL-6 expressed in horses with grade 4 EIPH than in horses with grade 0, 1, or 2 EIPH. At a high altitude, no difference was detected for IL-6 expression among the various EIPH grades. There was significantly greater overall expression of TNF-α mRNA at a high altitude; however, there was no difference within the various grades of EIPH. Expression of IL-10 was significantly affected by grade of EIPH because horses with grade 3 EIPH expressed significantly more IL-10 mRNA than did horses with grade 0 or 2 EIPH; this expression was not affected by location.

Conclusions and Clinical Relevance—At sea level, increased IL-6 expression was associated with more severe EIPH, and altitude may affect gene expressions of the proinflammatory cytokine TNF-α and anti-inflammatory cytokine IL-6. Studies on protein concentrations of cytokine expression are needed. The pathophysiologic importance of these findings remains to be explained.

Abstract

Objective—To determine concentrations of cytokine mRNA in horses with exercise-induced pulmonary hemorrhage (EIPH) after racing.

Animals—97 Thoroughbreds.

Procedures—Following tracheobronchoscopy, the severity of EIPH was graded (scale of 0 to 4), and venous blood samples were collected from 10 horses in each grade. After RNA isolation and cDNA synthesis, real-time PCR assay was conducted to detect cytokinespecific mRNA for interleukin (IL)-1, IL-6, and IL-10; interferon (INF)-γ; and tumor necrosis factor (TNF)-α.

Results—Neither location nor grade of EIPH affected the expression of IL-1 and INF-γ. There was significantly greater overall expression of IL-6 mRNA at sea level, with significantly more IL-6 expressed in horses with grade 4 EIPH than in horses with grade 0, 1, or 2 EIPH. At a high altitude, no difference was detected for IL-6 expression among the various EIPH grades. There was significantly greater overall expression of TNF-α mRNA at a high altitude; however, there was no difference within the various grades of EIPH. Expression of IL-10 was significantly affected by grade of EIPH because horses with grade 3 EIPH expressed significantly more IL-10 mRNA than did horses with grade 0 or 2 EIPH; this expression was not affected by location.

Conclusions and Clinical Relevance—At sea level, increased IL-6 expression was associated with more severe EIPH, and altitude may affect gene expressions of the proinflammatory cytokine TNF-α and anti-inflammatory cytokine IL-6. Studies on protein concentrations of cytokine expression are needed. The pathophysiologic importance of these findings remains to be explained.

Contributor Notes

Dr. Godfroid's present address is Section of Arctic Veterinary Medicine, Department of Food Safety and Infection Biology, Norwegian School of Veterinary Science, Tromso, Norway.

This manuscript represents a portion of a thesis submitted by the first author to the University of Pretoria Faculty of Veterinary Science as partial fulfillment of the requirements for a Doctor of Philosophy degree.

Supported by the Albert and Loraine Clay Endowment Fund for Support of Inter-Institutional Research Collaborations at the Maxwell H. Gluck Equine Research Center; and the Postgraduate Study Abroad Bursary Program, University of Pretoria, South Africa.

Presented in abstract form at the 25th American College of Veterinary Internal Medicine Forum, Seattle, June 2007.

Address correspondence to Dr. Saulez (montague.saulez@up.ac.za).
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