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Home / Equine Research Bank / Arthritis Res Ther / Carprofen inhibits the release of matrix metalloproteinases ...
Arthritis research & therapy2014; 15(6); R223; doi: 10.1186/ar4424

Carprofen inhibits the release of matrix metalloproteinases 1, 3, and 13 in the secretome of an explant model of articular cartilage stimulated with interleukin 1β.

Abstract: Arthritic diseases are characterized by the degradation of collagenous and noncollagenous extracellular matrix (ECM) components in articular cartilage. The increased expression and activity of matrix metalloproteinases (MMPs) is partly responsible for cartilage degradation. This study used proteomics to identify inflammatory proteins and catabolic enzymes released in a serum-free explant model of articular cartilage stimulated with the pro-inflammatory cytokine interleukin 1β (IL-1β). Western blotting was used to quantify the release of selected proteins in the presence or absence of the cyclooxygenase-2 specific nonsteroidal pro-inflammatory drug carprofen. Methods: Cartilage explant cultures were established by using metacarpophalangeal joints from horses euthanized for purposes other than research. Samples were treated as follows: no treatment (control), IL-1β (10 ng/ml), carprofen (100 μg/ml), and carprofen (100 μg/ml) + IL-1β (10 ng/ml). Explants were incubated (37°C, 5% CO2) over twelve day time courses. High-throughput nano liquid chromatography/mass spectrometry/mass spectrometry uncovered candidate proteins for quantitative western blot analysis. Proteoglycan loss was assessed by using the dimethylmethylene blue (DMMB) assay, which measures the release of sulfated glycosaminoglycans (GAGs). Results: Mass spectrometry identified MMP-1, -3, -13, and the ECM constituents thrombospondin-1 (TSP-1) and fibronectin-1 (FN1). IL-1β stimulation increased the release of all three MMPs. IL-1β also stimulated the fragmentation of FN1 and increased chondrocyte cell death (as assessed by β-actin release). Addition of carprofen significantly decreased MMP release and the appearance of a 60 kDa fragment of FN1 without causing any detectable cytotoxicity to chondrocytes. DMMB assays suggested that carprofen initially inhibited IL-1β-induced GAG release, but this effect was transient. Overall, during the two time courses, GAG release was 58.67% ± 10.91% (SD) for IL-1β versus 52.91% ± 9.35% (SD) with carprofen + IL-1β. Conclusions: Carprofen exhibits beneficial anti-inflammatory and anti-catabolic effects in vitro without causing any detectable cytotoxicity. Combining proteomics with this explant model provides a sensitive screening system for anti-inflammatory compounds.
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Publication Date: 2014-01-01 PubMed ID: 24373218PubMed Central: PMC3978949DOI: 10.1186/ar4424Google Scholar: Lookup
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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.

The study aims to portray how a drug, Carprofen, can reduce the release of certain proteins linked to arthritic diseases, specifically focusing on their effect in an explant model of articular cartilage.

Overview of the Research

  • The research focuses on arthritic diseases that degrade the extracellular matrix (ECM) components in the articular cartilage required for joints’ functionality. A primary cause of this degradation is the increased activity of matrix metalloproteinases (MMPs), a group of proteins.
  • To simulate the conditions of these diseases, the study used a cartilage explant model treated with Interleukin 1β (IL-1β), a pro-inflammatory cytokine responsible for cell signaling during inflammation.
  • These models were then treated with the drug Carprofen, a nonsteroidal anti-inflammatory drug, to observe its effect on the protein release.

Methodology of the Research

  • The research used the explant model were cartilage cultures established from metacarpophalangeal joints from horses.
  • These samples were then treated with IL-1β and Carprofen, often combined, over twelve days and observed for changes.
  • The study also used high-throughput nano liquid chromatography/mass spectrometry/mass spectrometry to identify potential proteins for western blot analysis, an experimental method used to detect specific proteins in a sample. Proteoglycan loss was measured using the dimethylmethylene blue (DMMB) assay.

Results of the Research

  • Interleukin 1β (IL-1β) treatment increased the release of MMP-1, -3, and -13 proteins within the model.
  • The study found that the application of Carprofen significantly decreased the release of these proteins. Despite its effectiveness, the inhibition of IL-1β-induced GAG release by Carprofen was transient.
  • There was also a decrease in the appearance of a 60 kDa fragment of FN1 (a glycoprotein involved in cell adhesion and migration processes involving cell surface interactions) with the use of Carprofen. Notably, Carprofen did not cause any detectable cytotoxicity to chondrocytes (cells found in healthy cartilage).

Conclusions of the Research

  • The study concluded that Carprofen showed beneficial anti-inflammatory and anti-catabolic effects in vitro (outside a living organism), meaning it could potentially be used to treat arthritic diseases.
  • The pairing of proteomics (the large-scale study of proteins) with this explant model provided a sensitive and efficient screening system for further research into anti-inflammatory compounds.

Cite This Article

APA
Williams A, Smith JR, Allaway D, Harris P, Liddell S, Mobasheri A. (2014). Carprofen inhibits the release of matrix metalloproteinases 1, 3, and 13 in the secretome of an explant model of articular cartilage stimulated with interleukin 1β. Arthritis Res Ther, 15(6), R223. https://doi.org/10.1186/ar4424

Publication

Arthritis research & therapy
ISSN: 1478-6362
NlmUniqueID: 101154438
Country: England
Language: English
Volume: 15
Issue: 6
Pages: R223

Researcher Affiliations

Williams, Adam
    Smith, Julia R
      Allaway, David
        Harris, Pat
          Liddell, Susan
            Mobasheri, Ali

              MeSH Terms

              • Animals
              • Anti-Inflammatory Agents, Non-Steroidal / pharmacology
              • Blotting, Western
              • Carbazoles / pharmacology
              • Cartilage, Articular / drug effects
              • Cartilage, Articular / enzymology
              • Chromatography, Liquid
              • Extracellular Matrix / metabolism
              • High-Throughput Screening Assays
              • Horses
              • Interleukin-1beta / pharmacology
              • Matrix Metalloproteinases / biosynthesis
              • Matrix Metalloproteinases / drug effects
              • Proteoglycans / metabolism
              • Tandem Mass Spectrometry

              Grant Funding

              • BB/G018030/1 / Biotechnology and Biological Sciences Research Council

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