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Src Kinase Regulates Nitric Oxide-induced Dedifferentiation and Cyc1ooxygenase-2 Expression in Articular Chondrocytes via p38 Kinase-dependent Pathway

  • Yu, Seon-Mi (Department of Biological Sciences, Kongju National University College of Natural Sciences) ;
  • Lee, Won-Kil (Department of Biological Sciences, Kongju National University College of Natural Sciences) ;
  • Yoon, Eun-Kyung (Department of Biological Sciences, Kongju National University College of Natural Sciences) ;
  • Lee, Ji-Hye (Department of Biological Sciences, Kongju National University College of Natural Sciences) ;
  • Lee, Sun-Ryung (Department of Biology, College of Natural Sciences, Cheju National University) ;
  • Kim, Song-Ja (Department of Biological Sciences, Kongju National University College of Natural Sciences)
  • Published : 2006.12.31
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Abstract

Background: Nitric oxide (NO) in articular chondrocytes regulates dedifferentiation and inflammatory responses by modulating MAP kinases. In this study, we investigated whether the Src kinase in chondrocytes regulates NO-induced dedifferentiation and cyclooxygenase-2 (COX-2) expression. Methods: Primary chondrocytes were treated with various concentrations of SNP for 24 h. The COX-2 and type II collagen expression levels were determined by immunoblot analysis, and prostaglandin $E_2\;(PGE_2)$ was determined by using a $PGE_2$ assay kit. Expression and distribution of p-Caveolin and COX-2 in rabbit articular chondrocytes and cartilage explants were determined by immunohistochemical staining and immunocytochemical staining, respectively. Results: SNP treatment stimulated Src kinase activation in a dose-dependent manner in articular chondrocytes. The Src kinase inhibitors PP2 [4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo(3,4-d)pyrimidine], a significantly blocked SNP-induced p38 kinase and caveolin-1 activation in a dose-dependent manner. Therefore, to determine whether Src kinase activation is associated with dedifferentiation and/or COX-2 expression and $PGE_2$ production. As expected, PP2 potentiated SNP-stimulated dedifferentiation, but completely blocked both COX-2 expression and $PGE_2$ production. And also, levels of p-Caveolin and COX-2 protein expression were increased in SNP-treated primary chondrocytes and osteoarthritic and rheumatoid arthritic cartilage, suggesting that p-Caveolin may playa role in the inflammatory responses of arthritic cartilage. Conclusion: Our previously studies indicated that NO caused dedifferentiation and COX-2 expression is regulated by p38 kinase through caveolin-1 (1). Therefore, our results collectively suggest that Src kinase regulates NO-induced dedifferentiation and COX-2 expression in chondrocytes via p38 kinase in association with caveolin-1.

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References

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