p38 Kinase Regulates Nitric Oxide-induced Dedifferentiation and Cyclooxygenase-2 Expression of Articular Chondrocytes
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Background: Caveolin, a family of integral membrane proteins are a principal component of caveolae membranes. In this study, we investigated the effect of p38 kinase on differentiation and on inflammatory responses in sodium nitroprusside (SNP)- treated chondrocytes. Methods: Rabbit articular chondrocytes were prepared from cartilage slices of 2-week-old New Zealand white rabbits by enzymatic digestion. SNP was used as a nitric oxide (NO) donor. In this experiments measuring SNP dose response, primary chondrocytes were treated with various concentrations of SNP for 24 h. The time course of the SNP response was determined by incubating cells with 1 mM SNP for the indicated time period (0∼24 h). The cyclooxygenase-2 (COX-2) and type II collagen expression levels were determined by immunoblot analysis, and prostaglandin E2 (PGE2) assay was used to measure the COX-2 activity. The tyrosine phosphorylation of caveolin-1 was determined by immunoblot analysis and immunostaining. Results: SNP treatment stimulated tyrosine phosphorylation of caveolin-1 and activation of p38 kinase. SNP additionally caused dedifferentiation and inflammatory response. We showed previously that SNP treatment stimulated activation of p38 kinase and ERK-1/-2. Inhibition of p38 kinase with SB203580 reduced caveolin-1 tyrosine phosphorylation and COX-2 expression but enhanced dedifferentiation, whereas inhibition of ERK with PD98059 did not affect caveolin-1 tyrosine phosphorylation levels, suggesting that ERK at least is not related to dedifferentiation and COX-2 expression through caveolin-1 tyrosine phosphorylation. Conclusion: Our results indicate that SNP in articular chondrocytes stimulates dedifferentiation and inflammatory response via p38 kinase signaling in association with caveolin-1 phosphorylation. (Immune Network 2006;6(3):117-122)
Abstract
Background: Caveolin, a family of integral membrane proteins are a principal component of caveolae membranes. In this study, we investigated the effect of p38 kinase on differentiation and on inflammatory responses in sodium nitroprusside (SNP)- treated chondrocytes. Methods: Rabbit articular chondrocytes were prepared from cartilage slices of 2-week-old New Zealand white rabbits by enzymatic digestion. SNP was used as a nitric oxide (NO) donor. In this experiments measuring SNP dose response, primary chondrocytes were treated with various concentrations of SNP for 24 h. The time course of the SNP response was determined by incubating cells with 1 mM SNP for the indicated time period (0∼24 h). The cyclooxygenase-2 (COX-2) and type II collagen expression levels were determined by immunoblot analysis, and prostaglandin E2 (PGE2) assay was used to measure the COX-2 activity. The tyrosine phosphorylation of caveolin-1 was determined by immunoblot analysis and immunostaining. Results: SNP treatment stimulated tyrosine phosphorylation of caveolin-1 and activation of p38 kinase. SNP additionally caused dedifferentiation and inflammatory response. We showed previously that SNP treatment stimulated activation of p38 kinase and ERK-1/-2. Inhibition of p38 kinase with SB203580 reduced caveolin-1 tyrosine phosphorylation and COX-2 expression but enhanced dedifferentiation, whereas inhibition of ERK with PD98059 did not affect caveolin-1 tyrosine phosphorylation levels, suggesting that ERK at least is not related to dedifferentiation and COX-2 expression through caveolin-1 tyrosine phosphorylation. Conclusion: Our results indicate that SNP in articular chondrocytes stimulates dedifferentiation and inflammatory response via p38 kinase signaling in association with caveolin-1 phosphorylation. (Immune Network 2006;6(3):117-122)
참고문헌 (33)
- "Differentiation of cartilage and bone", 1, pp.989~994 (1989)
- "Developmental patterns of cartilage", 4, pp.731~742 (1999)
- "Phenotypic stability of bovine articular chondrocytes after long-term culture in alginate beads", 107, pp.17~27 (1994)
- "Formation of nodular structures resembling mature articular cartilage in long-term primary cultures of human fetal epiphyseal chondrocytes on a hydrogel substrate", 37, pp.1338~1349 (1994)
- "Phenotypic modulation in sub-populations of human articular chondrocytes in vitro", 97, pp.361~371 (1990)
- "Articular cartilage and changes in arthritis", 3, pp.107~113 (2001)
- "The role of nitric oxide in articular cartilage breakdown in osteoarthritis", 10, pp.263~268 (1998)
- "Nitric oxide synthase and cyclooxygenases", 11, pp.202~209 (1999)
- "Nitric oxide and inflammatory mediators in the perpetuation of osteoarthritis", 3, pp.535~541 (2001)
- "ERK-1", 2 and p38 kinase oppositely regulate nitric oxide-induced apoptosis of chondrocytes in association with p53, caspase-3, and differentiation status, 2002, pp.277, 1332~1339
- "Protein kinase C-alpha and -zeta regulate nitric-oxide-induced NF-kappa B activation that mediates cyclooxygenase-2 expression and apoptosis but not dedifferentiation in articular chondrocytes", 303, pp.206~211 (2003)
- "Evidence for a regulated interaction between heterotrimeric G proteins and caveolin", 270, pp.15693~15701 (1995)
- "Src tyrosine kinases, G-alpha subunits, and H-Ras shear a common membrane-anchored scaffolding protein, caveolin.", 271, pp.29182~29190 (1996)
- "Co-purification and direct interaction of Ras with caveolin, an integral membrane protein of caveolae microdomains", 271, pp.9690~9697 (1996)
- "Caveolin versus calmodulin", 272, pp.25907~25912 (1997)
- "Caveolin, a family of scaffolding proteins for organizing "preassembled signaling complexes" at the plasma membrane", 273, pp.5419~5422 (1998)
- "Novel tyrosine kinase substrates from Rous sarcoma virus-transformed cells are present in the membrane skeleton", 108, pp.2401~2408 (1989)
- via activation of p38 mitogen-activated protein kinase and c-Src kinase, , 2001, 8094~8103
- "Opposing role of mitogen-activated protein kinase subtypes, Erk-1", -2 and p38, in the regulation of chondrogenesis of mesenchymes, 2000, pp.275, 5613~5619
- "Maintenance of differentiated phenotype of articular chondrocytes by protein kinase C and extracellular signal-regulated protein kinase", 277, pp.8412~8420 (2002)
- "Actin cytoskeletal architecture regulates nitric-oxide-induced apoptosis, dedifferentiation, and cyclooxygenase-2 exprssion in articular chondrocytes via mitogen-activated protein kinase and protei nkinase C pathways", 278, pp.42448~42456 (2003)
- "Insulin stimulates the tyrosine phosphorylation of caveolin", 129, pp.1523~1531 (1995)
- "Insulin-stimulated tyrosine phosphorylation of caveolin is specific for the differentiated adipocyte phenotype in 3T3-L1 cells", 272, pp.20706~20714 (1997)
- "Cholesterol depletion inhibitsepidermal growth factor receptor transactivation by angiotensin II in vascular smooth muscle cells", 276, pp.48269~48275 (2001)
- "Segregation of heterotrimeric G proteins in cell surface microdomains", 12, pp.685~698 (2001)
- "Targeting of nitric oxide synthase to endothelial cell caveolae via palmitoylation", 93, pp.6448~6453 (1996)
- "Endothelial nitric oxide synthase targeting to caveolae", 271, pp.22810~22814 (1996)
- "Localization of platelet-derived growth factor-stimulated phosphorylation cascade to caveolae", 271, pp.10299~10303 (1996)
- "Tyrosine phosphorylation of a 22-kDa protein is correlated with transformation by Rous sarcoma virus", 264, pp.20163~20166 (1989)
- "Early effects of pp60(v-src) kinase activation on caveolae", 71, pp.524~535 (1998)
- "Tyrosine-phosphorylated caveolin-1", 10, pp.975~986 (1999)
- "Caveolin-1 and a 29-kDa caveolin-associated protein are phosphorylated on tyrosine in cells expressing a temperature-sensitive v-Abl kinase", 266, pp.142~154 (2001)
- "p38 kinase-dependent and -independent inhibition of protein kinase C zeta and alpha regulates nitric- oxide-induced apoptosis and dedifferentiation of articular chondrocytes", 277, pp.30375~30381 (2002)