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.
Publication Date: 2003-06-28 PubMed ID: 12828249DOI: 10.2460/ajvr.2003.64.666Google Scholar: Lookup
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.
- 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
Researcher Affiliations
- Department of Large Animal Clinical Sciences, Michigan State University, East Lansing, MI 48824, USA.
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.- Carmona JU, Ríos DL, López C, Álvarez ME, Pérez JE. Proinflammatory and Anabolic Gene Expression Effects of Platelet-Rich Gel Supernatants on Equine Synovial Membrane Explants Challenged with Lipopolysaccharide. Vet Med Int 2017;2017:6059485.
- Carmona JU, Ríos DL, López C, Álvarez ME, Pérez JE, Bohórquez ME. In vitro effects of platelet-rich gel supernatants on histology and chondrocyte apoptosis scores, hyaluronan release and gene expression of equine cartilage explants challenged with lipopolysaccharide. BMC Vet Res 2016 Jul 1;12(1):135.
- Phitak T, Pothacharoen P, Kongtawelert P. Comparison of glucose derivatives effects on cartilage degradation. BMC Musculoskelet Disord 2010 Jul 15;11:162.
- Uitterlinden EJ, Koevoet JL, Verkoelen CF, Bierma-Zeinstra SM, Jahr H, Weinans H, Verhaar JA, van Osch GJ. Glucosamine increases hyaluronic acid production in human osteoarthritic synovium explants. BMC Musculoskelet Disord 2008 Sep 11;9:120.
- Kawasaki T, Kurosawa H, Ikeda H, Kim SG, Osawa A, Takazawa Y, Kubota M, Ishijima M. Additive effects of glucosamine or risedronate for the treatment of osteoarthritis of the knee combined with home exercise: a prospective randomized 18-month trial. J Bone Miner Metab 2008;26(3):279-87.
- Scotto d'Abusco A, Calamia V, Cicione C, Grigolo B, Politi L, Scandurra R. Glucosamine affects intracellular signalling through inhibition of mitogen-activated protein kinase phosphorylation in human chondrocytes. Arthritis Res Ther 2007;9(5):R104.
- Toegel S, Huang W, Piana C, Unger FM, Wirth M, Goldring MB, Gabor F, Viernstein H. Selection of reliable reference genes for qPCR studies on chondroprotective action. BMC Mol Biol 2007 Feb 26;8:13.
- Santangelo KS, Johnson AL, Ruppert AS, Bertone AL. Effects of hyaluronan treatment on lipopolysaccharide-challenged fibroblast-like synovial cells. Arthritis Res Ther 2007;9(1):R1.
- Gouze JN, Gouze E, Popp MP, Bush ML, Dacanay EA, Kay JD, Levings PP, Patel KR, Saran JP, Watson RS, Ghivizzani SC. Exogenous glucosamine globally protects chondrocytes from the arthritogenic effects of IL-1beta. Arthritis Res Ther 2006;8(6):R173.
- Nakamura H, Masuko K, Yudoh K, Kato T, Kamada T, Kawahara T. Effects of glucosamine administration on patients with rheumatoid arthritis. Rheumatol Int 2007 Jan;27(3):213-8.
- Lorenz W, Sigrist G, Shakibaei M, Mobasheri A, Trautmann C. A hypothesis for the origin and pathogenesis of rheumatoid diseases. Rheumatol Int 2006 May;26(7):641-54.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
