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Home / Equine Research Bank / Can J Vet Res / Detection and comparison of nitric oxide in clinically norma...
Canadian journal of veterinary research = Revue canadienne de recherche veterinaire1999; 63(4); 230-240;

Detection and comparison of nitric oxide in clinically normal horses and those with naturally acquired small intestinal strangulation obstruction.

Abstract: The purpose of this study was to determine whether nitric oxide (NO) is present in clinically normal horses under basal conditions and if it increases secondary to naturally acquired small intestinal strangulation obstruction. Thirty-one horses were used; 20 horses with naturally acquired small intestinal strangulation obstruction and 11 clinically normal horses with no signs of gastrointestinal tract disease. Jugular venous blood, abdominal fluid, and urine were collected for NO quantification. Plasma, abdominal fluid, and urine were stored at -70 degrees C until analyzed for NO using a chemiluminescent method. Biopsy specimens collected from the affected jejunal segment, during anesthesia or after immediately after euthanasia, or from the midjejunum of control horses, were divided into subsections for fixation in zinc formalin and cryopreservation in OCT gel. Nicotinamide adenine dinucleotide phosphate (reduced) (NADPH) diaphorase histochemical stains were performed on cryopreserved tissues and inducible nitric oxide synthase (iNOS) and nitrotyrosine immunohistochemical stains were performed on formalin-fixed, paraffin-embedded tissues. There were significantly greater plasma and abdominal fluid NO concentrations in affected horses as compared with controls, but there were no significant differences between horses for urine NO concentrations. There was a significant decrease in NADPH diaphorase stain in mucosal epithelium, vasculature, and leukocytes, and in submucosal plexi in affected horses compared with control horses. There was a significant increase in iNOS staining in mucosal and submucosal leukocytes and in mucosal leukocyte nitrotyrosine staining of the affected compared with control horses. Endothelial NOS and neuronal NOS are present under basal conditions in the jejunum of horses and probably mediate physiologic or cytoprotective effects. Plasma and abdominal fluid, but not urine, NO concentrations increase subsequent to small intestinal strangulation obstruction; this may be associated with increased mucosal and submucosal iNOS staining in leukocytes, which was likely due to increased expression subsequent to stimuli associated with ischemia. The increased nitrotyrosine staining in mucosal leukocytes of affected horses likely reflects the presence of peroxynitrite subsequent to increased NO and superoxide production and may reflect a cytotoxic role of NO in small intestinal strangulation obstruction in horses.
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Publication Date: 1999-10-26 PubMed ID: 10534001PubMed Central: PMC1189558
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study investigates whether nitric oxide (NO) levels increase in horses suffering from small intestinal strangulation obstruction compared to healthy horses. The findings show that NO concentrations in plasma and abdominal fluid do indeed rise in affected horses, implying a role of NO in the pathological changes seen in this condition.

Methodology

  • The study involved 31 horses, out of which 20 suffered from naturally acquired small intestinal strangulation obstruction and the remaining 11 were healthy.
  • Biological samples including blood, abdominal fluid, and urine were collected from each horse for NO quantification.
  • The samples were stored at -70 degrees C and analyzed with a chemiluminescent method for NO.
  • Additionally, biopsy specimens from the affected midjejunum area (part of the small intestine) were collected during anesthesia or immediately after euthanasia.
  • The biopsy specimens were processed for histochemical and immunohistochemical stains to detect NADPH diaphorase, inducible nitric oxide synthase (iNOS), and nitrotyrosine.

Findings

  • Among the affected horses, significantly higher NO concentrations were found in the plasma and abdominal fluid compared to the control group, suggesting a possible role of NO in intestinal obstruction.
  • There were no significant differences in urine NO concentrations between the two groups.
  • A significant decrease in NADPH diaphorase stain was observed in the mucosal epithelium, vasculature, and leukocytes, as well as in submucosal plexi of the affected horses when compared with the control horses.
  • A significant increase in iNOS staining was observed in mucosal and submucosal leukocytes, suggesting increased iNOS activity following ischemic injury.
  • Increased nitrotyrosine staining, a marker of oxidative/nitrosative stress, was also observed in mucosal leukocytes of the affected horses, indicating potential cytotoxicity of NO in cases of obstruction.

Interpretation

  • Endothelial NO synthase (eNOS) and neuronal NOS (nNOS), both of which produce NO, were found to be present under basal conditions in the jejunum of horses and likely play a role in normal physiological processes.
  • The increase in plasma and abdominal fluid NO, but not urine NO, following small intestinal strangulation obstruction could be associated with increased iNOS activity in leukocytes, likely due to ischemic stimuli.
  • Increased nitrotyrosine staining in mucosal leukocytes of affected horses could hint at the formation of peroxynitrite through NO and superoxide production, pointing towards a possible cytotoxic role of NO in this condition.

Cite This Article

APA
Mirza MH, Oliver JL, Seahorn TL, Hosgood G, Moore RM. (1999). Detection and comparison of nitric oxide in clinically normal horses and those with naturally acquired small intestinal strangulation obstruction. Can J Vet Res, 63(4), 230-240.

Publication

Canadian journal of veterinary research = Revue canadienne de recherche veterinaire
ISSN: 0830-9000
NlmUniqueID: 8607793
Country: Canada
Language: English
Volume: 63
Issue: 4
Pages: 230-240

Researcher Affiliations

Mirza, M H
  • Department of Veterinary Clinical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge 70803, USA.
Oliver, J L
    Seahorn, T L
      Hosgood, G
        Moore, R M

          MeSH Terms

          • Animals
          • Enzyme Induction
          • Female
          • Horse Diseases / metabolism
          • Horses
          • Intestinal Mucosa / chemistry
          • Intestinal Obstruction / metabolism
          • Intestinal Obstruction / veterinary
          • Male
          • Nitric Oxide / analysis
          • Nitric Oxide / pharmacology
          • Nitric Oxide Synthase / metabolism
          • Reference Values

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          Citations

          This article has been cited 5 times.
          1. Borunova SF, Tkachev N, Iolchiev B, Artyushina Z, Abramov P, Nikitina M, Silanteva A, Khusnetdinova N, Serezhenkov V. Estimation of Nitrite-Nitric Oxide Derivative-In Horses with Intestinal Colic by ESR Spectroscopy.. Vet Sci 2020 Nov 29;7(4).
            doi: 10.3390/vetsci7040191pubmed: 33260335google scholar: lookup
          2. Camelo JS Jr, Martins AR, Rosa E, Ramos SG, Hehre D, Bancalari E, Suguihara C. Angiotensin II type 1 receptor blockade partially attenuates hypoxia-induced pulmonary hypertension in newborn piglets: relationship with the nitrergic system.. Braz J Med Biol Res 2012 Feb;45(2):163-71.
            doi: 10.1590/s0100-879x2012007500014pubmed: 22310488google scholar: lookup
          3. Mirza MH, Seahorn TL, Oliver JL, Hosgood G, Moore RM. Detection and comparison of nitric oxide in clinically healthy horses and those with naturally acquired strangulating large colon volvulus.. Can J Vet Res 2005 Apr;69(2):106-15.
            pubmed: 15971674
          4. Hubert JD, Seahorn TL, Klei TR, Hosgood G, Horohov DW, Moore RM. Clinical signs and hematologic, cytokine, and plasma nitric oxide alterations in response to Strongylus vulgaris infection in helminth-naïve ponies.. Can J Vet Res 2004 Jul;68(3):193-200.
            pubmed: 15352544
          5. Hubert JD, Seahorn TL, Klei TR, Hosgood G, Moore RM. Temporal effects of freezing on plasma nitric oxide concentrations in ponies.. Can J Vet Res 2003 Jan;67(1):72-4.
            pubmed: 12528834
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