Effects of glucosamine and chondroitin sulfate on mediators of osteoarthritis in cultured equine chondrocytes stimulated by use of recombinant equine interleukin-1beta.
OBJECTIVE
To determine whether glucosamine and chondroitin sulfate (CS) at concentrations approximating those achieved in plasma by oral administration would influence gene expression of selected mediators of osteoarthritis in cytokine-stimulated equine articular chondrocytes.


METHODS
Samples of grossly normal articular cartilage obtained from the metacarpophalangeal joint of 13 horses.


METHODS
Equine chondrocytes in pellet culture were stimulated with a subsaturating dose of recombinant equine interleukin (reIL)-1beta. Effects of prior incubation with glucosamine (2.5 to 10.0 microg/mL) and CS (5.0 to 50.0 microg/mL) on gene expression of matrix metalloproteinase (MMP)-1, -2, -3, -9, and -13; aggrecanase 1 and 2; inducible nitric oxide synthase (iNOS); cyclooxygenase (COX)-2; nuclear factor kappaB; and c-Jun-N-terminal kinase (JNK) were assessed by use of a quantitative real-time polymerase chain reaction assay.


RESULTS
Glucosamine at a concentration of 10 microg/mL significantly reduced reIL-1beta-induced mRNA expression of MMP-13, aggrecanase 1, and JNK. Reductions in cytokine-induced expression were also observed for iNOS and COX-2. Chondroitin sulfate had no effect on gene expression at the concentrations tested.


CONCLUSIONS
Concentrations of glucosamine similar to those achieved in plasma after oral administration in horses exerted pretranslational regulation of some mediators of osteoarthritis, an effect that may contribute to the cartilage-sparing properties of this aminomonosaccharide. Analysis of results of this study indicated that the influence of CS on pretranslational regulation of these selected genes is limited or lacking.
Neil
Kirsten M
KM
Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing 48824, USA.
Orth
Michael W
MW
Coussens
Paul M
PM
Chan
Pooi-See
PS
Caron
John P
JP
D051722:ADAM Proteins / Q000096:biosynthesis / Q000235:genetics; D000071121:ADAMTS4 Protein; D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D019902:Chondrocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D002809:Chondroitin Sulfates / Q000494:pharmacology*; D051546:Cyclooxygenase 2 / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D005944:Glucosamine / Q000494:pharmacology*; D006734:Horse Diseases / Q000188:drug therapy*; D006736:Horses; D007375:Interleukin-1 / Q000494:pharmacology*; D048670:MAP Kinase Kinase 4 / Q000096:biosynthesis / Q000235:genetics; D020782:Matrix Metalloproteinases / Q000096:biosynthesis / Q000235:genetics; D016328:NF-kappa B / Q000096:biosynthesis / Q000235:genetics; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis / Q000235:genetics; D010003:Osteoarthritis / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000662:veterinary*; D011348:Procollagen N-Endopeptidase / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary
D007375:Interleukin-1; D016328:NF-kappa B; D011994:Recombinant Proteins; D002809:Chondroitin Sulfates; D052247:Nitric Oxide Synthase Type II; D051546:Cyclooxygenase 2; D048670:MAP Kinase Kinase 4; D051722:ADAM Proteins; D020782:Matrix Metalloproteinases; D011348:Procollagen N-Endopeptidase; D000071121:ADAMTS4 Protein; D005944:Glucosamine