Evaluation of various compounds to inhibit activity of matrix metalloproteinases in the tear film of horses with ulcerative keratitis.
Abstract: To examine in vitro effects of various antiproteolytic compounds on activity of matrix metalloproteinase (MMP)-2 and -9 in the tear film of horses with active corneal ulcers. Methods: Samples of tear film obtained from the eyes of 34 horses with active ulcerative keratitis. Methods: Horses were sedated, and tear samples were collected from the lower fornix of 34 ulcerated eyes by use of capillary tubes. The protease inhibitors 0.2% EDTA, 0.1% doxycycline, 10% N-acetylcysteine (NAC), 0.1% solution of a modified dipeptide that contains hydroxamic acid (ie, ilomostat), 0.1% alpha1-proteinase inhibitor (PI), 0.5% alpha1-PI, and 100% fresh equine serum (ES) were used to treat pooled samples. Amount of latent and active MMP-2 and -9 was measured by optical density scanning of gelatin zymograms of treated and untreated tear samples. Results: Pooled tear samples obtained from ulcerated eyes contained the latent and active forms of MMP-2 and -9. Compared with MMP activity in untreated samples, total MMP activity (sum of all bands detected) observed on the gelatin zymogram gels was reduced by 99.4% by EDTA, 96.3% by doxycycline, 98.8% by NAC, 98.9% by ilomostat, 52.4% by 0.1% alpha1-PI, 93.6% by 0.5% alpha1-PI, and 90.0% by ES. Conclusions: We documented that EDTA, doxycycline, NAC, ilomostat, alpha1PI, and ES inhibited MMP activity in vitro. Because these compounds use different mechanisms to inhibit various families of proteases in the tear film of horses, a combination of these protease inhibitors may be beneficial for treatment of corneal ulcers in horses.
Publication Date: 2003-09-19 PubMed ID: 13677383DOI: 10.2460/ajvr.2003.64.1081Google Scholar: Lookup
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- Journal Article
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The research article analyzes the effects of antiproteolytic compounds on the activity of certain enzymes in the tear film of horses with corneal ulcers.
Overview of the Study
- The study was designed to examine the in vitro effects of several antiprotelytic compounds on the activity of matrix metalloproteinase (MMP)-2 and -9. These enzymes are present in the tear film formulations of horses suffering from active corneal ulcers.
Methodology
- The tear film samples for this study were acquired from 34 horses diagnosed with active ulcerative keratitis.
- These horses were sedated during sample collection. The samples were gathered from the lower fornix of their ulcerated eyes using capillary tubes.
- The compounds that were used to treat the pooled samples included 0.2% EDTA, 0.1% doxycycline, 10% N-acetylcysteine (NAC), 0.1% solution of modified dipeptide ilomostat, 0.1% alpha1-proteinase inhibitor (PI), 0.5% alpha1-PI, and 100% fresh equine serum (ES).
- To measure the quantity of latent and active MMP-2 and -9, the researchers performed optical density scanning of gelatin zymograms of the treated and non-treated tear samples.
Findings of the Study
- The researchers discovered that the pooled tear samples obtained from ulcerated eyes held both latent and active forms of MMP-2 and -9.
- Compared to the MMP activity in the untreated samples, total MMP activity was substantially reduced in the treated samples. EDTA reduced activity by 99.4%, Doxycycline by 96.3%, NAC by 98.8%, Ilomostat by 98.9%, 0.1% alpha1-PI by 52.4%, 0.5% alpha1-PI by 93.6% and ES by 90.0%.
Conclusions
- The study concluded that the tested compounds – EDTA, doxycycline, NAC, ilomostat, alpha1PI, and ES – were successful in inhibiting the MMP activity in vitro.
- Since these compounds work differently to inhibit various families of proteases in the tear film of horses, combining these protease inhibitors could potentially improve the treatment of corneal ulcers in horses.
Cite This Article
APA
Ollivier FJ, Brooks DE, Kallberg ME, Komaromy AM, Lassaline ME, Andrew SE, Gelatt KN, Stevens GR, Blalock TD, van Setten GB, Schultz GS.
(2003).
Evaluation of various compounds to inhibit activity of matrix metalloproteinases in the tear film of horses with ulcerative keratitis.
Am J Vet Res, 64(9), 1081-1087.
https://doi.org/10.2460/ajvr.2003.64.1081 Publication
Researcher Affiliations
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA.
MeSH Terms
- Acetylcysteine / pharmacology
- Animals
- Corneal Ulcer / enzymology
- Corneal Ulcer / veterinary
- Doxycycline / pharmacology
- Edetic Acid / pharmacology
- Electrophoresis / veterinary
- Horse Diseases / enzymology
- Horses
- Hydroxamic Acids / pharmacology
- Image Processing, Computer-Assisted
- Matrix Metalloproteinase 2 / metabolism
- Matrix Metalloproteinase 9 / metabolism
- Matrix Metalloproteinase Inhibitors
- Protease Inhibitors / pharmacology
- Tears / enzymology
- alpha 1-Antitrypsin / pharmacology
Grant Funding
- EY05587 / NEI NIH HHS
Citations
This article has been cited 14 times.- van Setten GB. Cellular Stress in Dry Eye Disease-Key Hub of the Vicious Circle. Biology (Basel) 2024 Aug 28;13(9).
- Marchegiani A, Gialletti R, Cassarani MP, Cerquetella M, Attili AR, Lombardo G, Lombardo M, Spaterna A, Arcelli R. Riboflavin/UV-A corneal phototherapy as stand-alone management of ulcerative keratitis in dogs. Vet Med (Praha) 2022 Apr;67(4):190-198.
- Meissner JM, Chmielińska A, Ofri R, Cisło-Sankowska A, Marycz K. Extracellular Vesicles Isolated from Equine Adipose-Derived Stromal Stem Cells (ASCs) Mitigate Tunicamycin-Induced ER Stress in Equine Corneal Stromal Stem Cells (CSSCs). Curr Issues Mol Biol 2024 Apr 9;46(4):3251-3277.
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- Donovan C, Koudouna E, Margo CE, Avila MY, Espana EM. Genipin Delays Corneal Stromal Enzymatic Digestion. Transl Vis Sci Technol 2021 Aug 2;10(9):25.
- Chen X, Wang Y, Tian J, Shao Y, Zhu B, Wang J, Hua Z. Quantitative Chemical Proteomics Reveals Resveratrol Inhibition of A549 Cell Migration Through Binding Multiple Targets to Regulate Cytoskeletal Remodeling and Suppress EMT. Front Pharmacol 2021;12:636213.
- Farghali HA, AbdElKader NA, AbuBakr HO, Ramadan ES, Khattab MS, Salem NY, Emam IA. Corneal Ulcer in Dogs and Cats: Novel Clinical Application of Regenerative Therapy Using Subconjunctival Injection of Autologous Platelet-Rich Plasma. Front Vet Sci 2021;8:641265.
- Briffod C, Bédard C, Vanore M. Stability of epidermal growth factor, fibronectin, and alpha-2-macroglobulin in canine serum under different storage conditions. Can J Vet Res 2019 Oct;83(4):267-271.
- Galera PD, Brooks DE. Optimal management of equine keratomycosis. Vet Med (Auckl) 2012;3:7-17.
- Martin de Bustamante MG, Good KL, Leonard BC, Hollingsworth SR, Edwards SG, Knickelbein KE, Cooper AE, Thomasy SM, Maggs DJ. Medical management of deep ulcerative keratitis in cats: 13 cases. J Feline Med Surg 2019 Apr;21(4):387-393.
- Sebbag L, Showman L, McDowell EM, Perera A, Mochel JP. Impact of Flow Rate, Collection Devices, and Extraction Methods on Tear Concentrations Following Oral Administration of Doxycycline in Dogs and Cats. J Ocul Pharmacol Ther 2018 Jul Aug;34(6):452-459.
- Hazra S, Guha R, Jongkey G, Palui H, Mishra A, Vemuganti GK, Basak SK, Mandal TK, Konar A. Modulation of matrix metalloproteinase activity by EDTA prevents posterior capsular opacification. Mol Vis 2012;18:1701-11.
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