Abstract
The mechanism of endothelin-1 (ET-1)-induced nitric oxide (NO) production, MMP-1 production and MMP-13 production was investigated in human osteoarthritis chondrocytes. The cells were isolated from human articular cartilage obtained at surgery and were cultured in the absence or presence of ET-1 with or without inhibitors of protein kinase or LY83583 (an inhibitor of soluble guanylate cyclase and of cGMP). MMP-1, MMP-13 and NO levels were then measured by ELISA and Griess reaction, respectively. Additionally, inducible nitric oxide synthase (iNOS) and phosphorylated forms of p38 mitogen-activated protein kinase, p44/42, stress-activated protein kinase/Jun-N-terminal kinase and serine-threonine Akt kinase were determined by western blot. Results show that ET-1 greatly increased MMP-1 and MMP-13 production, iNOS expression and NO release. LY83583 decreased the production of both metalloproteases below basal levels, whereas the inhibitor of p38 kinase, SB202190, suppressed ET-1-stimulated production only. Similarly, the ET-1-induced NO production was partially suppressed by the p38 kinase inhibitor and was completely suppressed by the protein kinase A kinase inhibitor KT5720 and by LY83583, suggesting the involvement of these enzymes in relevant ET-1 signalling pathways. In human osteoarthritis chondrocytes, ET-1 controls the production of MMP-1 and MMP-13. ET-1 also induces NO release via iNOS induction. ET-1 and NO should thus become important target molecules for future therapies aimed at stopping cartilage destruction.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aminoquinolines / pharmacology
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Apoptosis / drug effects
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Carbazoles / pharmacology
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Cartilage, Articular / pathology*
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Cells, Cultured / drug effects
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Cells, Cultured / metabolism
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Chondrocytes / drug effects*
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Chondrocytes / metabolism
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Collagenases / biosynthesis*
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Collagenases / genetics
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Cyclic AMP-Dependent Protein Kinase Type II
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Cyclic AMP-Dependent Protein Kinases / antagonists & inhibitors
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Cyclic GMP / antagonists & inhibitors
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Endothelin-1 / pharmacology*
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Enzyme Induction / drug effects
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Female
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Guanylate Cyclase / antagonists & inhibitors
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Humans
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Imidazoles / pharmacology
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Indoles / pharmacology
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MAP Kinase Signaling System / drug effects
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Male
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Matrix Metalloproteinase 1 / biosynthesis
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Matrix Metalloproteinase 1 / genetics
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Matrix Metalloproteinase 13
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Middle Aged
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Nitric Oxide / biosynthesis*
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Nitric Oxide Synthase Type II / biosynthesis
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Nitric Oxide Synthase Type II / genetics
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Osteoarthritis, Knee / metabolism
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Osteoarthritis, Knee / pathology*
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Phosphorylation / drug effects
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Protein Processing, Post-Translational / drug effects
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Pyridines / pharmacology
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Pyrroles / pharmacology
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Signal Transduction / drug effects
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p38 Mitogen-Activated Protein Kinases / antagonists & inhibitors
Substances
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Aminoquinolines
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Carbazoles
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Endothelin-1
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Imidazoles
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Indoles
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Pyridines
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Pyrroles
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Nitric Oxide
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KT 5720
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6-anilino-5,8-quinolinedione
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Nitric Oxide Synthase Type II
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Cyclic AMP-Dependent Protein Kinase Type II
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Cyclic AMP-Dependent Protein Kinases
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p38 Mitogen-Activated Protein Kinases
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Collagenases
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MMP13 protein, human
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Matrix Metalloproteinase 13
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Matrix Metalloproteinase 1
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Guanylate Cyclase
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Cyclic GMP
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4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole