Nitric oxide production by equine articular cells in vitro.
Recent research in several species has suggested nitric oxide (NO) as a mediator of articular cartilage damage and an inhibitor of cartilage matrix neosynthesis. This study investigated NO production by cultured equine articular chondrocytes in response to 2 arthritogenic molecules, namely lipopolysaccharide (LPS) and interleukin-1 beta (IL-1 beta), and compared NO production by cultured equine synoviocytes stimulated with LPS. Synoviocytes exhibited a low basal level of NO synthesis (measured as nitrite, a NO metabolite) that was neither significantly increased nor decreased by exposure to LPS. Basal NO synthesis by synoviocytes was not significantly reduced by competitive inhibitors of nitric oxide synthase (NOS). In contrast, chondrocytes treated with LPS or IL-1 beta synthesised nitrite in a dose-related manner. Inhibitors of NOS suppressed nitrite production to below the basal levels of release of unstimulated cells. Dexamethasone, an inhibitor of induction of the inducible isoform of NOS (iNOS), reduced nitrite synthesis by LPS-stimulated chondrocytes. Western blot analysis revealed expression, in response to LPS, of protein in the same molecular weight range as iNOS identified in other species. This work demonstrates that equine chondrocytes have the capacity to synthesise NO, although its exact roles in cartilage metabolism have yet to be determined.
Frean
S P
SP
Department of Veterinary Basic Sciences, Royal Veterinary College, North Mymms, Herts, UK.
Bryant
C E
CE
Fröling
I L
IL
Elliott
J
J
Lees
P
P
D000818:Animals; D015153:Blotting, Western / Q000379:methods / Q000662:veterinary; D002358:Cartilage, Articular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005938:Glucocorticoids / Q000494:pharmacology; D006736:Horses / Q000378:metabolism*; D007375:Interleukin-1 / Q000494:pharmacology; D007536:Isomerism; D008070:Lipopolysaccharides / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D013583:Synovial Membrane / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors
D005938:Glucocorticoids; D007375:Interleukin-1; D008070:Lipopolysaccharides; D009573:Nitrites; D009569:Nitric Oxide; D003907:Dexamethasone; D019001:Nitric Oxide Synthase