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American journal of veterinary research2003; 64(6); 666-671; doi: 10.2460/ajvr.2003.64.666

Influence of glucosamine on matrix metalloproteinase expression and activity in lipopolysaccharide-stimulated equine chondrocytes.

Abstract: To characterize potential mechanisms of action of glucosamine inhibition of matrix metalloproteinase (MMP) expression and activity in lipopolysaccharide (LPS)-stimulated equine chondrocytes. Methods: Chondrocytes cultured from samples of metacarpophalangeal articular cartilage collected from cadaveric limbs of horses. Methods: The effect of glucosamine on MMP activity in conditioned medium from LPS-stimulated cartilage explants was determined by a colorimetric assay with azocoll substrate. Treatments consisted of negative and positive controls, glucose (50 mM), and glucosamine (50, 25, 6.25, 3, and 1.5 mM). The influence of glucosamine on MMP synthesis was determined in chondrocytes in pellet culture incubated with LPS (20 microg/mL). Concentration of MMP-13 was quantified in spent medium via ELISA; nonspecific MMP activity was determined via azocoll digestion in organomercurial-activated medium. Effects of glucosamine on MMP mRNA concentration in similarly treated chondrocytes were determined by northern blot hybridization with MMP-1, -3, and -13 probes. Statistical analyses were performed with 2-way ANOVA. Results: Glucosamine had no effect on activated MMP activity but inhibited MMP protein expression, as determined by azocoll digestion (glucosamine, 3 to 50 mM) and MMP-13 ELISA (glucosamine, 1.5 to 50 mM). Resting mRNA concentrations for MMP-1, -3, and -13 mRNA were significantly lower in cultures exposed to glucosamine at concentrations of 50 and 25 mM than those of positive controls. Conclusions: Glucosamine appears capable of pretranslational, and possibly also translational, regulation of MMP expression; data suggest a potential mechanism of action for chondroprotective effects of this aminomonosaccharide.
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Publication Date: 2003-06-28 PubMed ID: 12828249DOI: 10.2460/ajvr.2003.64.666Google Scholar: Lookup
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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 how glucosamine, a common dietary supplement, inhibits the expression and activity of matrix metalloproteinase (MMP) proteins in equine cartilage cells that are exposed to lipopolysaccharides. The findings suggest that glucosamine may protect cartilage by regulating MMP expression prior to and perhaps during protein translation.

Research Methodology

  • The researchers cultured chondrocytes, which are cartilage cells, from the metacarpophalangeal articular cartilage collected from horse cadaver limbs.
  • This cultured cartilage was exposed to lipopolysaccharides (LPS), a type of molecule that stimulates an immune response.
  • The impact of glucosamine on the activity of matrix metalloproteinase (MMP) in the medium in which the cartilage was cultured was then determined using a colorimetric assay with azocoll substrate.
  • The treatments evaluated were negative and positive controls, glucose, and varying concentrations of glucosamine.
  • In a similar setup, the chondrocytes in pellet culture were incubated with LPS to determine the influence of glucosamine on MMP synthesis.
  • The concentration of a type of MMP, MMP-13, was measured in the medium using ELISA, and the general MMP activity was determined using azocoll digestion.
  • The effects of glucosamine on the concentration of MMP mRNA in comparable treatment scenarios were measured by northern blot hybridization using MMP-1, -3, and -13 probes.

Results and Conclusion

  • The results showed that glucosamine did not affect the activity of already activated MMP but it did inhibit MMP protein expression, as found by azocoll digestion and MMP-13 ELISA tests.
  • The resting mRNA concentrations of MMP-1, -3, and -13 were significantly lower in cultures exposed to high concentrations of glucosamine (50 and 25 mM) compared to the positive controls.
  • Therefore, it was concluded that glucosamine might be capable of regulating the expression of MMP at the pretranslational level, and potentially also at the translational level.
  • This proposes a possible mechanism through which glucosamine could exert chondroprotective effects, preserving the stability and health of cartilage.

Cite This Article

APA
Byron CR, Orth MW, Venta PJ, Lloyd JW, Caron JP. (2003). Influence of glucosamine on matrix metalloproteinase expression and activity in lipopolysaccharide-stimulated equine chondrocytes. Am J Vet Res, 64(6), 666-671. https://doi.org/10.2460/ajvr.2003.64.666

Publication

American journal of veterinary research
ISSN: 0002-9645
NlmUniqueID: 0375011
Country: United States
Language: English
Volume: 64
Issue: 6
Pages: 666-671

Researcher Affiliations

Byron, Christopher R
  • Department of Large Animal Clinical Sciences, Michigan State University, East Lansing, MI 48824, USA.
Orth, Michael W
    Venta, Patrick J
      Lloyd, James W
        Caron, John P

          MeSH Terms

          • Animals
          • Chondrocytes / drug effects
          • Chondrocytes / enzymology
          • Chondrocytes / metabolism
          • Gene Expression Regulation, Enzymologic / drug effects
          • Glucosamine / pharmacology
          • Horses
          • Lipopolysaccharides / pharmacology
          • Matrix Metalloproteinases / biosynthesis
          • Matrix Metalloproteinases / genetics
          • Matrix Metalloproteinases / metabolism
          • RNA, Messenger / genetics
          • RNA, Messenger / metabolism

          Citations

          This article has been cited 11 times.
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