FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
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.
Get Full Text
Publication Date: 1999-10-26 PubMed ID: 10534001PubMed Central: PMC1189558
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • 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

          References

          This article includes 31 references
          1. Rakestraw PC, Snyder JR, Woliner MJ, Sanders KM, Shuttleworth CW. Involvement of nitric oxide in inhibitory neuromuscular transmission in equine jejunum.. Am J Vet Res 1996 Aug;57(8):1206-13.
            pubmed: 8836376
          2. Dawson TM, Bredt DS, Fotuhi M, Hwang PM, Snyder SH. Nitric oxide synthase and neuronal NADPH diaphorase are identical in brain and peripheral tissues.. Proc Natl Acad Sci U S A 1991 Sep 1;88(17):7797-801.
            pubmed: 1715581doi: 10.1073/pnas.88.17.7797google scholar: lookup
          3. Moncada S, Higgs EA. Endogenous nitric oxide: physiology, pathology and clinical relevance.. Eur J Clin Invest 1991 Aug;21(4):361-74.
            pubmed: 1718757doi: 10.1111/j.1365-2362.1991.tb01383.xgoogle scholar: lookup
          4. Meschter CL, Craig D, Hackett R. Histopathological and ultrastructural changes in simulated large colonic torsion and reperfusion in ponies.. Equine Vet J 1991 Nov;23(6):426-33.
            pubmed: 1778159doi: 10.1111/j.2042-3306.1991.tb03755.xgoogle scholar: lookup
          5. Prichard M, Ducharme NG, Wilkins PA, Erb HN, Butt M. Xanthine oxidase formation during experimental ischemia of the equine small intestine.. Can J Vet Res 1991 Oct;55(4):310-4.
            pubmed: 1790484
          6. Kubes P, Hunter J, Granger DN. Ischemia/reperfusion-induced feline intestinal dysfunction: importance of granulocyte recruitment.. Gastroenterology 1992 Sep;103(3):807-12.
            pubmed: 1323498doi: 10.1016/0016-5085(92)90010-vgoogle scholar: lookup
          7. Ischiropoulos H, Zhu L, Chen J, Tsai M, Martin JC, Smith CD, Beckman JS. Peroxynitrite-mediated tyrosine nitration catalyzed by superoxide dismutase.. Arch Biochem Biophys 1992 Nov 1;298(2):431-7.
            pubmed: 1416974doi: 10.1016/0003-9861(92)90431-ugoogle scholar: lookup
          8. Snyder JR, Pascoe JR, Olander HJ, Harmon FA, Hinds DM, Vatistasa NJ, Tyler WS. Ultrastructural mucosal injury after experimental ischemia of the ascending colon in horses.. Am J Vet Res 1992 Oct;53(10):1917-24.
            pubmed: 1456541
          9. Kubes P. Ischemia-reperfusion in feline small intestine: a role for nitric oxide.. Am J Physiol 1993 Jan;264(1 Pt 1):G143-9.
            pubmed: 8430797doi: 10.1152/ajpgi.1993.264.1.G143google scholar: lookup
          10. Archer S. Measurement of nitric oxide in biological models.. FASEB J 1993 Feb 1;7(2):349-60.
            pubmed: 8440411doi: 10.1096/fasebj.7.2.8440411google scholar: lookup
          11. Beckman JS, Crow JP. Pathological implications of nitric oxide, superoxide and peroxynitrite formation.. Biochem Soc Trans 1993 May;21(2):330-4.
            pubmed: 8395426doi: 10.1042/bst0210330google scholar: lookup
          12. Moncada S, Higgs A. The L-arginine-nitric oxide pathway.. N Engl J Med 1993 Dec 30;329(27):2002-12.
            pubmed: 7504210doi: 10.1056/NEJM199312303292706google scholar: lookup
          13. Kanwar S, Tepperman BL, Payne D, Sutherland LR, Kubes P. Time course of nitric oxide production and epithelial dysfunction during ischemia/reperfusion of the feline small intestine.. Circ Shock 1994 Mar;42(3):135-40.
            pubmed: 8025978
          14. Horne MM, Pascoe PJ, Ducharme NG, Barker IK, Grovum WL. Attempts to modify reperfusion injury of equine jejunal mucosa using dimethylsulfoxide, allopurinol, and intraluminal oxygen.. Vet Surg 1994 Jul-Aug;23(4):241-9.
            pubmed: 8091626doi: 10.1111/j.1532-950x.1994.tb00478.xgoogle scholar: lookup
          15. Wilkins PA, Ducharme NG, Lowe JE, Schwark WS, Meschter C, Erb HN. Measurements of blood flow and xanthine oxidase activity during postischemic reperfusion of the large colon of ponies.. Am J Vet Res 1994 Aug;55(8):1168-77.
            pubmed: 7978659
          16. Moore RM, Bertone AL, Bailey MQ, Muir WW, Beard WL. Neutrophil accumulation in the large colon of horses during low-flow ischemia and reperfusion.. Am J Vet Res 1994 Oct;55(10):1454-63.
            pubmed: 7998704
          17. Ribbons KA, Zhang XJ, Thompson JH, Greenberg SS, Moore WM, Kornmeier CM, Currie MG, Lerche N, Blanchard J, Clark DA. Potential role of nitric oxide in a model of chronic colitis in rhesus macaques.. Gastroenterology 1995 Mar;108(3):705-11.
            pubmed: 7533111doi: 10.1016/0016-5085(95)90442-5google scholar: lookup
          18. Moore RM, Muir WW, Granger DN. Mechanisms of gastrointestinal ischemia-reperfusion injury and potential therapeutic interventions: a review and its implications in the horse.. J Vet Intern Med 1995 May-Jun;9(3):115-32.
            pubmed: 7674213doi: 10.1111/j.1939-1676.1995.tb03285.xgoogle scholar: lookup
          19. Adams HR. Physiologic, pathophysiologic, and therapeutic implications for endogenous nitric oxide.. J Am Vet Med Assoc 1996 Oct 1;209(7):1297-302.
            pubmed: 8837656
          20. Beckman JS, Koppenol WH. Nitric oxide, superoxide, and peroxynitrite: the good, the bad, and ugly.. Am J Physiol 1996 Nov;271(5 Pt 1):C1424-37.
            pubmed: 8944624doi: 10.1152/ajpcell.1996.271.5.C1424google scholar: lookup
          21. McCafferty DM, Mudgett JS, Swain MG, Kubes P. Inducible nitric oxide synthase plays a critical role in resolving intestinal inflammation.. Gastroenterology 1997 Mar;112(3):1022-7.
            pubmed: 9041266doi: 10.1053/gast.1997.v112.pm9041266google scholar: lookup
          22. Kooreman K, Babbs C, Fessler J. Effect of ischemia and reperfusion on oxidative processes in the large colon and jejunum of horses.. Am J Vet Res 1998 Mar;59(3):340-6.
            pubmed: 9522955
          23. White NA, Moore JN, Trim CM. Mucosal alterations in experimentally induced small intestinal strangulation obstruction in ponies.. Am J Vet Res 1980 Feb;41(2):193-8.
            pubmed: 7369591
          24. Hsu SM, Raine L, Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures.. J Histochem Cytochem 1981 Apr;29(4):577-80.
            pubmed: 6166661doi: 10.1177/29.4.6166661google scholar: lookup
          25. Palmer RM, Ferrige AG, Moncada S. Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor.. Nature 1987 Jun 11-17;327(6122):524-6.
            pubmed: 3495737doi: 10.1038/327524a0google scholar: lookup
          26. Freeman DE, Cimprich RE, Richardson DW, Gentile DG, Orsini JA, Tulleners EP, Fetrow JP. Early mucosal healing and chronic changes in pony jejunum after various types of strangulation obstruction.. Am J Vet Res 1988 Jun;49(6):810-8.
            pubmed: 3400918
          27. Flaherty JT, Weisfeldt ML. Reperfusion injury.. Free Radic Biol Med 1988;5(5-6):409-19.
            pubmed: 3076884doi: 10.1016/0891-5849(88)90115-3google scholar: lookup
          28. MacDonald MH, Pascoe JR, Stover SM, Meagher DM. Survival after small intestine resection and anastomosis in horses.. Vet Surg 1989 Nov-Dec;18(6):415-23.
            pubmed: 2603371doi: 10.1111/j.1532-950x.1990.tb01116.xgoogle scholar: lookup
          29. Aoki N, Johnson G 3rd, Lefer AM. Beneficial effects of two forms of NO administration in feline splanchnic artery occlusion shock.. Am J Physiol 1990 Feb;258(2 Pt 1):G275-81.
            pubmed: 2305894doi: 10.1152/ajpgi.1990.258.2.G275google scholar: lookup
          30. Kubes P, Suzuki M, Granger DN. Nitric oxide: an endogenous modulator of leukocyte adhesion.. Proc Natl Acad Sci U S A 1991 Jun 1;88(11):4651-5.
            pubmed: 1675786doi: 10.1073/pnas.88.11.4651google scholar: lookup
          31. Moncada S, Palmer RM, Higgs EA. Nitric oxide: physiology, pathophysiology, and pharmacology.. Pharmacol Rev 1991 Jun;43(2):109-42.
            pubmed: 1852778

          Citations

          This article has been cited 6 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
          6. Hajimohammadi A, Ghane M, Ghari Tehrani M, Paravar B, Mirzaei A, Razavi S, Nikzad M. Association of the severity of colic in horses with oxidative stress biomarkers, acute-phase proteins, and certain trace elements. J Equine Sci 2023 Sep;34(3):73-81.
            doi: 10.1294/jes.34.73pubmed: 37781567google scholar: lookup
          Subscribe to our newsletter
          Join 99,727 horse owners like you who receive our email updates on horse health and nutrition.