Expression of cyclooxygenase genes in the jejunum of horses during low-flow ischemia and reperfusion

Hugo Hilton, Jorge Nieto, Peter F Moore, Faye A. Harmon, Diane K. Naydan, Jack R. Snyder

Research output: Contribution to journalArticle

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Abstract

Objective-To determine expression of cyclooxygenase (COX) genes 1 and 2 (also called prostaglandin-endoperoxide synthases 1 and 2) and stability of housekeeping gene expression during low-flow ischemia and reperfusion in the jejunum of horses. Animals-5 healthy adult horses. Procedures-Horses were anesthetized, and two 30-cm segments of jejunum were surgically exteriorized. Blood flow was maintained at baseline (untreated) values in 1 (control) segment and was decreased to 20% of baseline (low-flow ischemia) for 75 minutes, followed by 75 minutes of reperfusion, in the other (experimental) segment. Biopsy samples were collected from experimental segments at baseline (T0), after 75 minutes of ischemia (T1), and after 75 minutes of reperfusion (T2); samples were collected from control segments at T0 and T2. Horses were euthanized 24 hours after induction of ischemia (T3), and additional samples were collected. Samples were evaluated histologically. Total RNA was extracted; expression of COX genes and stability of 8 housekeeping genes were determined via quantitative real-time PCR assays. Results-COX-1 and COX-2 genes were constitutively expressed in baseline samples. Low-flow ischemia resulted in significant upregulation of COX-2 gene expression at each subsequent time point, compared with baseline values. The most stably expressed reference genes were β-actin and hypoxanthine phosphoribosyltransferase, whereas glyceralde-hyde 3-phosphate dehydrogenase and β-2 microglobulin were the least stably expressed. Conclusions and Clinical Relevance-Low-flow ischemia resulted in upregulation of COX-2 gene expression in the jejunum of horses. Housekeeping genes traditionally used as internal standards may not be stable in this tissue during arterial low-flow ischemia and reperfusion.

Original languageEnglish (US)
Pages (from-to)681-686
Number of pages6
JournalAmerican Journal of Veterinary Research
Volume72
Issue number5
DOIs
StatePublished - May 2011

Fingerprint

prostaglandin synthase
Jejunum
Prostaglandin-Endoperoxide Synthases
ischemia
jejunum
Horses
Reperfusion
Ischemia
Gene Expression
horses
Cyclooxygenase 2
Essential Genes
genes
Cyclooxygenase 1
gene expression
normal values
sampling
Up-Regulation
Hypoxanthine Phosphoribosyltransferase
hypoxanthine

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Expression of cyclooxygenase genes in the jejunum of horses during low-flow ischemia and reperfusion. / Hilton, Hugo; Nieto, Jorge; Moore, Peter F; Harmon, Faye A.; Naydan, Diane K.; Snyder, Jack R.

In: American Journal of Veterinary Research, Vol. 72, No. 5, 05.2011, p. 681-686.

Research output: Contribution to journalArticle

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abstract = "Objective-To determine expression of cyclooxygenase (COX) genes 1 and 2 (also called prostaglandin-endoperoxide synthases 1 and 2) and stability of housekeeping gene expression during low-flow ischemia and reperfusion in the jejunum of horses. Animals-5 healthy adult horses. Procedures-Horses were anesthetized, and two 30-cm segments of jejunum were surgically exteriorized. Blood flow was maintained at baseline (untreated) values in 1 (control) segment and was decreased to 20{\%} of baseline (low-flow ischemia) for 75 minutes, followed by 75 minutes of reperfusion, in the other (experimental) segment. Biopsy samples were collected from experimental segments at baseline (T0), after 75 minutes of ischemia (T1), and after 75 minutes of reperfusion (T2); samples were collected from control segments at T0 and T2. Horses were euthanized 24 hours after induction of ischemia (T3), and additional samples were collected. Samples were evaluated histologically. Total RNA was extracted; expression of COX genes and stability of 8 housekeeping genes were determined via quantitative real-time PCR assays. Results-COX-1 and COX-2 genes were constitutively expressed in baseline samples. Low-flow ischemia resulted in significant upregulation of COX-2 gene expression at each subsequent time point, compared with baseline values. The most stably expressed reference genes were β-actin and hypoxanthine phosphoribosyltransferase, whereas glyceralde-hyde 3-phosphate dehydrogenase and β-2 microglobulin were the least stably expressed. Conclusions and Clinical Relevance-Low-flow ischemia resulted in upregulation of COX-2 gene expression in the jejunum of horses. Housekeeping genes traditionally used as internal standards may not be stable in this tissue during arterial low-flow ischemia and reperfusion.",
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