[en] OBJECTIVES: To determine the effects of two drugs, N-monomethyl-L-arginine (L-NMMA) and N-acetylcysteine (NAC), on interleukin-1beta (IL-1beta), nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production by human chondrocytes. The effect of aceclofenac (ACECLO), a non-steroidal antiinflammatory drug (NSAID), was also examined. METHODS: Human chondrocytes were enzymatically isolated from osteoarthritic knee cartilage and then maintained in culture in suspension for 48h in the absence or in the presence of lipopolysaccharide (LPS) (10 microg/ml), L-NMMA (0.5mM), NAC (1mM) or ACECLO (6.10(-6)M). IL-1beta and PGE(2) productions were quantified by specific immunoassays. Nitrite was measured in the culture supernatants by a spectrophotometric method based upon the Griess reaction. Cyclooxygenase-2 (COX-2), inducible NO synthase (iNOS) and IL-1beta gene expressions were quantified by transcription of mRNA followed by real time and quantitative polymerase chain reaction. COX-2 protein expression was analysed by Western blot. RESULTS: LPS markedly increased the expression of IL-1beta, iNOS and COX-2 genes. In parallel, NO(2) and PGE(2) amounts found in the culture supernatants were significantly enhanced whereas IL-1beta was immunologically undetectable. The addition of L-NMMA (0.5mM) fully blocked LPS-induced NO production but greatly increased PGE(2) production, suggesting a negative effect of NO on PGE(2) synthesis. Inversely, NO production was stimulated by NAC while PGE(2) production was not affected. Interestingly, NAC increased the IL-1beta and iNOS mRNA levels but did not significantly modify COX-2 mRNA expression. L-NMMA did not significantly affect the expression of IL-1beta, iNOS and COX-2. The amount of COX-2 protein did not change in the presence of the antioxidants. Finally, ACECLO fully blocked the production of PGE(2) by chondrocytes without affecting the levels of COX-2 mRNA. CONCLUSIONS: The stimulation of IL-1beta, NO and PGE(2) production by LPS is differentially controlled by reactive oxygen species (ROS). In fact, L-NMMA and NAC have different mechanisms of action on the regulation of NO and PGE(2) productions. L-NMMA fully inhibits NO but increases PGE(2) production whereas NAC up-regulates NO but does not modify PGE(2) synthesis. The stimulating effect of L-NMMA on PGE(2) production is not controlled at the transcriptional level. These findings suggest that antioxidant therapy could have different effects according to the oxygen radical species targeted.
Disciplines :
General & internal medicine
Author, co-author :
Mathy, Marianne ; Université de Liège - ULiège > Département des sciences de la motricité > Unité de recherche sur l'os et le cartillage (U.R.O.C.)
Deby, Ginette ; Université de Liège - ULiège > Centre de l'oxygène : Recherche et développement (C.O.R.D.)
Reginster, Jean-Yves ; Université de Liège - ULiège > Département des sciences de la santé publique > Epidémiologie et santé publique
Ayache, N.
Pujol, J*-P
Henrotin, Yves ; Université de Liège - ULiège > Département des sciences de la motricité > Unité de recherche sur l'os et le cartillage (U.R.O.C.) - Didactique des sciences de la santé - Pathologie générale et physiopathologie
Language :
English
Title :
Regulation by reactive oxygen species of interleukin-1beta, nitric oxide and prostaglandin E(2) production by human chondrocytes.
Publication date :
2002
Journal title :
Osteoarthritis and Cartilage
ISSN :
1063-4584
eISSN :
1522-9653
Publisher :
W.B. Saunders, London, United Kingdom
Volume :
10
Issue :
7
Pages :
547-55
Peer reviewed :
Peer Reviewed verified by ORBi
Commentary :
Copyright 2002 OsteoArthritis Research Society International. Published by Elsevier Science Ltd. All rights reserved.
Tiku M.L., Liesch J.B., Robertson F.M. (1990) Production of hydrogen peroxide by rabbit articular chondrocytes. J. Immunol 145:690-696.
Henrotin Y., Deby-Dupont G., Deby C., De Bruyn M., Lamy M., Franchimont P. (1993) Production of active oxygen species by isolated human chondrocytes. Br. J. Rheumatol 32:562-567.
Stadler J., Stefanovic-Racic M., Billiar T.R., Curran R.D., Mcintyre L.A., Georgescu H.I., Simmons R.L., Evans C.H. (1991) Articular chondrocytes synthesize nitric oxide in response to cytokines and lipopolysaccharide. J. Immunol 147:3915-3920.
Pelletier J.P., Lascau-Coman V., Jovanovic D., Fernandes J.C., Manning P., Connor J.R., Currie M.G., Martel-Pelletier J. (1999) Selective inhibition of inducible nitric oxide synthase in experimental osteoarthritis is associated with reduction in tissue levels of catabolic factors. J. Rheumatol 26:2002-2014.
Sasaki K., Hattori T., Fujisawa T., Takahashi K., Inoue H., Takigawa M. (1998) Nitric oxide mediates Interleukin-1-induced gene expression of matrix metalloproteinases and basic fibroblast growth factor in cultured rabbit articular chondrocytes. J. Biochem 123:431-439.
Taskiran D., Stefanovic-Racic M., Georgescu H., Evans C. (1994) Nitric oxide mediates suppression of cartilage proteoglycan synthesis by Interleukin-1. Biochem. Biophysics. Res. Commun 200:142-148.
Pelletier J.P., Mineau F., Ranger P., Tardif G., Martel-Pelletier J. (1996) The increased synthesis of inducible nitric oxide inhibits IL-1ra synthesis by human articular chondrocytes: Possible role in osteoarthritic cartilage degradation. Osteoarth. Cart 4:77-84.
Corbett J.A., Kwon G., Turk J., McDaniel M.L. (1993) IL-1β induces the coexpression of both nitric oxide synthase and cyclooxygenase by islets of Langerhans: Activation of cyclooxygenase by nitric oxide. Biochem 32:13767-13770.
Salvemini D., Misko T.P., Masferrer J.L., Seibert K., Currie M.G., Needleman P. (1993) Nitric oxide activates cyclooxygenase enzymes. Proc. Natl. Acad. Sci 90:7240-7244.
Tetsuka T., Daphna-Iken D., Miller B.W., Guan Z., Baier L.D., Morrison A.R. (1996) Nitric oxide amplifies interleukin 1-induced cyclooxygenase-2 expression in rat mesangial cells. J. Clin. Invest 97:2051-2056.
Habib A., Bernard C., Lebret M., Creminon C., Esposito B., Tedgui A. (1997) Regulation of the expression of cyclooxygenase-2 by nitric oxide in rat peritoneal macrophages. J. Immunol 158:3845-3851.
Amin A.R., Attur M., Patel R.N., Thakker G.D., Marshall P.J., Rediske J. (1997) Superinduction of cyclooxygenase-2 activity in human osteoarthritis-affected cartilage. J. Clin. Invest 99:1231-1237.
Henrotin Y.E., Zheng S.X., Deby G.P., Labasse A.H., Crielaard J.M.R., Reginster J.Y.L. (1998) Nitric oxide down-regulates interleukin 1β (IL-1β) stimulated IL-6, IL-8, and prostaglandin E2 production by human chondrocytes. J. Rheumatol 25:1595-1601.
Attur M.G., Patel R.N., Patel P.D., Abramson S.B., Amin A.R. (1999) Tetracycline up-regulates COX-2 expression and prostaglandin E2 production independent of its effect on nitric oxide. J. Immunol 162:3160-3167.
Stadler J., Harbrecht B., Di Silvio M., Curran R.D., Jordan M.L., Simmons R.L., Billiar T.R. (1993) Endogenous nitric oxide inhibits the synthesis of cyclooxygenase products and interleukin-6 by rat Kupffer cells. J. Leuko. Biol 53:165-172.
Clancy R., Leszczynska-Piziak J., Abramson S. (1992) Nitric oxide, an endothelial cell relaxation factor, inhibits neutrophil superoxide anion production via a direct action on the NADPH oxidase. J. Clin. Invest 90:1116-1121.
Ginn-Pease M.E., Whisler R.L. (1998) Redox signals and NF-κB activation in T cells. Free Radic. Biol. Med 25:346-361.
Ortega Mateo A., de Artinano A.A. (2000) Nitric oxide reactivity and mechanisms involved in its biological effects. Pharmacol. Res 42:421-427.
Green L., Wagner D., Glogowski J., Skipper P., Wishnok J., Tannenbaum S. (1982) Analysis of nitrate, nitrite, and (15N) nitrate in biological fluids. Anal. Biochem 126:131-138.
Gilliam M.B., Sherman M.P., Griscavage J.M., Ignarro L.J. (1993) A Spectrophotometric assay for nitrate using NADPH oxidation by aspergillus nitrate reductase. Anal. Biochem 212:359-365.
Serteyn D., Deby-Dupont G., Pincemail J., Mottart E., Philippart C., Lamy M. (1988) Equine postanaesthetic myositis. Thromboxanes, prostacyclin and prostaglandin E2 production. Vet. Res. Commun 12:219-226.
Labarca A., Paigen K. (1980) A simple, rapid and sensitive DNA procedure. Anal. Biochem 102:344-354.
Patel R., Attur M.G., Dave M.N., Kumar S., Lee J.C., Abramson S.B. (1999) Regulation of nitric oxide and prostaglandin E2 production by CSAID™ (SB203580) in murine macrophages and bovine chondrocytes stimulated with LPS. Inflamm. Res 48:337-343.
MacMillan-Crow L.A., Thompson J.A. (1999) Tyrosine modifications and inactivation of active site manganese superoxide dismutase mutant (Y34F) by peroxynitrite. Arch. Biochem. Biophys 366:82-88.
Roy B., Lepoivre M., Henry Y., Fontecave M. (1995) Inhibition of ribonucleotide reductase by nitric oxide derived from thionitrites: Reversible modifications of both subunits. Biochemistry 34:5411-5418.
Goodwin D.C., Landino L.M., Marnett L.J. (1999) Effects of nitric oxide and nitric oxide-derived species on prostaglandin endoperoxide synthase and prostaglandin biosynthesis. FASEB J 13:1121-1136.
Kosonen O., Kankaanranta H., Malo-Ranta U., Ristimaki A., Moilanen E. (1998) Inhibition by nitric oxide-releasing compounds of prostacyclin production in human endothelial cells. Br. J. Pharmacol 125:247-254.
Martinez J., Sanchez T., Moreno J.J. (1999) Regulation of prostaglandin E2 production by the superoxide radical and nitric oxide in mouse peritoneal macrophages. Free Rad. Res 32:303-311.
Jiang B., Brecher P. (2000) N-acetyl-L-cysteine potentiates interleukin-1 beta induction of nitric oxide synthase: Role of p44/42 mitogen-activated protein kinases. Hypertension 35:914-918.
Huang A., Vita J.A., Venema R.C., Keaney J.F. Jr. (2000) Ascorbic acid enhances endothelial nitric-oxide synthase activity by increasing intracellular tetrahydrobiopterin. J. Biol. Chem 275:17399-17406.
Rao G.H., Tate M.R., Murthy M., Hebbel R.P., White J.G. (1994) Influence of antioxidants on arachidonic acid metabolism and platelet function. Biochem. Med. Metab. Biol 51:74-79.
Henrotin Y., Mouithys-Mickalad A., Mathy-Hartert M., Zheng S.X., Deby G., Reginster J.Y. (2000) The antioxidant properties of non steroidal anti-inflammatory drugs. Osteoarthritis Cartilage , [Abstract]; 8(SUPPL. B).
Henrotin Y., de Leval X., Mathy-Hartert M., Mouithys-Mickalad A., Deby G., Dogné J.-M. (2001) In vitro effects of aceclofenac and its metabolites on the production by chondrocytes of inflammatory mediators. Inflamm. Res 50:391-399.
