Skip to main content
Download PDF
  • Published:

Gelatinolytic Activity in Tracheal Aspirates of Horses with Chronic Obstructive Pulmonary Disease

Acta Veterinaria Scandinavica volume 38, pages 17–27 (1997)Cite this article

Abstract

The gelatinolytic activity in tracheal aspirates (TA) of horses with chronic obstructive pulmonary disease (COPD) was analyzed using SDS-PAGE-gelatin-gel electrophoresis (zymography) and compared to TAs from healthy controls. The 110-90 kD MMP-9 type gelatinase was high in symptomatic disease phases (permanent disease 0.46 ±0.15, p<0.001; or intermittent disease 0.47 ±0.12, p<0.001) compared to healthy controls (0.10 ± 0.07). Similarly, the overall gelatinolytic activity, the activity in high-mw gelatinolytic bands (210-190 and 150 kD) and in proteolytically processed fragments in the 50-40 kD range were high, whereas the 75-65 kD MMP-2 was not altered. These findings suggest that MMP-9 type gelatinases, originating possibly from neutrophils or macrophages, and products thereof have a role in the pathogenesis of equine respiratory diseases, whereas MMP-2 type gelatinases represent house-keeping proteinases involved with normal tissue remodelling. The gelatinolytic activity in TAs correlated with the β-glucuronidase activity, which indicates that they are simultaneously elevated in the respiratory secretions of horses suffering from COPD and might both be of same origin, or have a causal relationship.

References

  • Beech J: Tracheobronchial aspirates. In: Beech J. (ed.): Equine respiratory disorders. Lea and Febi- ger, Philadelphia, USA. 1991, pp 41–53.

    Google Scholar 

  • Burrell MH: Endoscopic and virological observations on respiratory disease in a group of young Thoroughbred horses in training. Equine vet. J. 1985, 17, 99–103.

    Article  CAS  Google Scholar 

  • Campbell EJ, Cury JD, Shapiro SD, Goldberg GI, Welgus HG: Neutral proteinases of human mononuclear phagocytes. Cellular differentiation markedly alters cell phenotype for serine proteinases, metalloproteinases, and tissue inhibitor of metalloproteinases. J. Immunol. 1991, 146, 1286–1293.

    CAS  PubMed  Google Scholar 

  • Campbell EJ, Senior RM, Welgus HG: Extracellular- matrix injury during lung inflammation. Chest 1987, 92, 161–167.

    Article  CAS  Google Scholar 

  • Clarke AF: Chronic pulmonary disease - A multifaceted disease complex in the horse. Ir. vet. J. 1987, 41, 258–264.

    Google Scholar 

  • Collier IE, Wilhelm SM, Eisen AZ, Marmer BL, Grant GA, Selzer JL, Kronberger AHC, Bauer EA, Goldberg GI: H-ras oncogene-transformed human bronchial epithelial cells (TBE-1) secrete a single metalloprotease capable of degrading basement membrane collagen. J. Biol. Chem. 1988, 263, 6579–6587.

    CAS  PubMed  Google Scholar 

  • Deegen E: Das chronisch lungenkranke Pferd und sein Einsatz im Sport. (Horse with a chronic lung disease and its input in sport.). Praktische Tier- artz 1987, 65, 15–19.

    Google Scholar 

  • Deegen E, Klein H-K: Interpleural pressure measurement and bronchial spasmolysis tests in the horse performed with transportable oesophageal pressure measuring instrument. Pferdeheilkunde 1987, 3, 213–221.

    Article  Google Scholar 

  • Derksen FJ: Chronic obstructive pulmonary disease. In: Beech, J. (ed.): Equine respiratory disorders, Lea & Febiger, Philadelphia, USA, 1991, 223–235.

    Google Scholar 

  • Derksen F, Arthur RM, Dillon G, Pascoe J, Sweeney C: A round table discussion on chronic obstructive pulmonary disease (COPD). Equine Practice 1991, 73, 25–28.

    Google Scholar 

  • Desrivieres S, Lu H, Peyri N, Soria C, Legrand Y, Menashi S: Activation of the 92 kDa type IV collagenase by tissue kallikrein. J. Cell. Physiol. 1993, 157, 587–593.

    Article  CAS  Google Scholar 

  • Garbisa S, Ballin M, Daga-Gordini D, Fastelli G, Naturale M, Negro A, Semenzato G, Liotta LA: Transient expression of type IV collagenolytic metalloproteinase by human mononuclear phagocytes. J. Biol. Chem. 1986, 261, 2369–2375.

    CAS  PubMed  Google Scholar 

  • Griinig G: Proteaseaktivitat im Tracheobronchial- sekret von Pferden mit COPD: Pathophysiologi- sche Bedeutung. (Protease activity in tracheobronchial secretions of horses with COPD and its relevance for pathogenesis.). Pferdeheilkunde 1985, 7, 55–63.

    Article  Google Scholar 

  • Guttmann I, Bergmeyer HU: Determination of urea with glutamate dehydrogenase as indicator enzyme. In: Bergmayer, HU (ed.): Methods of enzymatic analysis. 2nd ed. Academic Press, New York. 1974, pp. 1794–1798.

    Google Scholar 

  • Hibbs DS, Hembry RM, Docherty APJ, Reynolds JJ: Purification and characterization of human 72- kDa gelatinase (type IV collagenase). Use of immunolocalization to demonstrate the noncoordinate regulation of the 72-kDa and 95-kDa gelatinases by human fibroblasts. Biol. Chem. Hoppe-Seyler 1991, 372, 287–296.

    Google Scholar 

  • Hibbs MS, Hoidal JR, Kang AH: Expression of a metalloproteinase that degrades native type V collagen and denatured collagens by cultured human alveolar macrophages. J. Clin. Invest. 1987, 80, 1644–1650.

    Article  CAS  Google Scholar 

  • Kalebic T, Garbisa S, Glaser B, Liotta LA: Basement membrane collagen: Degradation by migrating endothelial cells. Science 1983, 227, 281–283.

    Article  Google Scholar 

  • Koivunen A-L, Maisi P, Fang W, Sandholm M: Inhibition of the protease activity in tracheobronchial aspirates of horses with COPD. Amer. J. vet. Res. 1996, 57, 603–607.

    CAS  Google Scholar 

  • Koj A, Chudnik J, Dubin A: Substrate specificity and modification of the active center of elastase-like neutral proteinases from horse blood leucocytes. Biochem. J. 1976, 753, 397–402.

    Article  Google Scholar 

  • Kolodny EH, Mumford RA: Human leucocyte acid hydrolases: Characterization of eleven lysosomal enzymes and study of reaction conditions for their automated analysis. Clin. Chim. Acta 1976, 70, 247–257.

    Article  CAS  Google Scholar 

  • Konttinen YT, Lindy O, Suomalainen K, Ritchlin C, Saari H, Vauhkonen M, Lauhio A, Santavirta S, Sorsa T: Substrate specificity and activation mechanisms of collagenase from human rheumatoid synovium. Matrix 1991, 77, 395–403.

    Article  Google Scholar 

  • Kusukawa J, Sasaguri Y, Shima I, Kameyama T, Morimatsu M: Production of matrix metalloproteinase 2 (gelatinase/ type IV collagenase) and 3 (stromelysin) by cultured oral squamous cell carcinoma. J. Oral Pathol. 1992, 21, 221–224.

    Article  CAS  Google Scholar 

  • Lauhio A, Konttinen YT, Tschesche H, Nordstrdm D, Salo T, Lahdevirta J, Golub LM, Sorsa T: Reduction of matrix metalloproteinase-8 levels during long-term doxycycline treatment. Antimicrob. Agents and Chemother. 1994a, 38, 400–402.

    Article  CAS  Google Scholar 

  • Lauhio A, Salo T, Ding Y, Konttinen YT, Nordstrdm D, Tschesche H, Lahdevirta J, Golub LM, Sorsa T: In vivo inhibition of human neutrophil collagenase (MMP-8) activity during long-term combination therapy of doxycycline and NS AID in acute reactive arthritis. Clin. Exp. Immunol. 1994b, 98, 21–28.

    Article  CAS  Google Scholar 

  • MacNamara B, Bauer S, Iafe J: Endoscopic evaluation of exercise-induced pulmonary haemorrhage and chronic obstructive pulmonary disease in association with poor performance in racing Stan- dardbreds. J. Amer. vet. med. Assoc. 1990, 196, 443–445.

    CAS  Google Scholar 

  • Mainardi CL, Pourmotable TF, Hasty KA: Inflammatory phagocytes and connective tissue degrading metalloproteinases. Amer. J. med. Sci. 1991, 302, 171–175.

    Article  CAS  Google Scholar 

  • Maisi P, Koivunen A-L, Rantala A-R, Turgut K: 3- glucuronidase and trypsin inhibitor capacity of tracheal lavage fluid as indicators of seasonal airway irritation in the horse. Equine vet. J. 1994, 26, 385–391.

    Article  CAS  Google Scholar 

  • Moore BR, Krakowka S, Robertson JT, Cummings JM: Cytologic evaluation of bronchoalveolar lavage fluid obtained from Standardbred racehorses with inflammatory airway disease. Amer. J. vet. Res. 1995, 56, 562–567.

    CAS  Google Scholar 

  • Rennard S, Basset G, Lecossier D, Donnel K, Martin P, Crystal RG: Estimation of volume of epithelial lining fluid recovered by lavage by using urea as a marker of dilution. J. Appl. Physiol. 1986, 60, 532–538.

    Article  CAS  Google Scholar 

  • Rossdale PD, Hopes R, Wingfield Digby NJ, Offord K: Epidemiological study of wastage among racehorses 1982 and 1983. Vet. Rec. 1985, 116, 66–69.

    Article  CAS  Google Scholar 

  • Saari H, Suomalainen K, Lindy O, Konttinen YT, Sorsa T: Activation of latent human neutrophil collagenase by reactive oxygen species and serine proteases. Biochem. Biophys. Res. Com- mun. 1990, 171, 979–987.

    Article  CAS  Google Scholar 

  • Sandholm M, Jdrvinen M, Halonen S, Eriksson A, Nyholm K, Maisi P: Enzymology of tracheal lavage fluid in horses with bronchial hyperreactivity. In: Proceedings of the International Society for Animal Clinical Biochemistry. Ed: J.J. Kaneko, Davis, California, USA 1990, pp. 51–157.

  • Senior RM, Griffin GL, Fliszar CJ, Shapiro SD, Goldberg GI, Welgus HG: Human 92- and 72-kilodal- ton type IV collagenases are elastases. J. Biol. Chem. 1991, 266, 7870–7875.

    CAS  PubMed  Google Scholar 

  • Sepper R, Konttinen YT, Sorsa T, Koski H: Gelatino- lytic and type IV collagenolytic activity in bronchiectasis disease. Chest 1994, 106, 1129–1133.

    Article  CAS  Google Scholar 

  • Sibille Y, Reynolds HY: Macrophages and polymorphonuclear neutrophils in lung defense and injury. Am. Rev. Respir. Dis. 1990, 141, 471–501.

    Article  CAS  Google Scholar 

  • Stockley RA: Proteolytic enzymes, their inhibitors and lung diseases. Clinical Science 1983, 64, 119–126.

    Article  CAS  Google Scholar 

  • Van den Ingh TSGAM: Morphological aspects of bronchitis and bronchiolitis in the horse. In: Deegen E, Beadle RE. Hippiatrika (eds.): Proceedings of the international symposium of lung function and respiratory diseases in the horse. Germany. 1985, pp. 13–15.

  • Weiss SJ: Tissue destruction by neutrophils. New Engl. J. Med. 1989, 320, 365–376.

    CAS  Google Scholar 

  • Woessner JF: Matrix metalloproteinases and their inhibitors in connective tissue remodelling. FASEB J. 1991, 5, 2145–2154.

    Article  CAS  Google Scholar 

Download references

Acknowledgment

This study was supported by the University of Veterinary Medicine, Emil Aaltonen foundation, the Academy of Finland and Astra Research Foundation.

Author information

Authors and Affiliations

  1. Department of Clinical Veterinary Sciences, Faculty of Veterinary Medicine, University of Helsinki, P.O. Box 57, Hämeentie 57, FIN-00014, Helsinki, Finland

    A.-L. Koivunen, P. Maisi & M. Sandholm

  2. Institute of Biomedicine, Department of Anatomy, University of Helsinki, and Division of Rheumatic Disease, Fourth Department of Medicine, University Central Hospital, Helsinki, Finland

    Y. T. Konttinen

Authors
  1. A.-L. Koivunen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. Maisi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y. T. Konttinen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Sandholm
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to P. Maisi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Koivunen, AL., Maisi, P., Konttinen, Y.T. et al. Gelatinolytic Activity in Tracheal Aspirates of Horses with Chronic Obstructive Pulmonary Disease. Acta Vet Scand 38, 17–27 (1997). https://doi.org/10.1186/BF03548504

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1186/BF03548504

Keywords

Acta Veterinaria Scandinavica

ISSN: 1751-0147