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Differential mechanism of NF-κB inhibition by two glucocorticoid receptor modulators in rheumatoid arthritis synovial fibroblasts

Valerie Gossye (UGent) , Dirk Elewaut (UGent) , Nadia Bougarne (UGent) , Debby Bracke (UGent) , Serge Van Calenbergh (UGent) , Guy Haegeman (UGent) and Karolien De Bosscher (UGent)
(2009) ARTHRITIS AND RHEUMATISM. 60(11). p.3241-3250
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Abstract
Objective. To investigate and compare the molecular mechanisms by which 2 glucocorticoid receptor (GR)-activating compounds, dexamethasone (DEX) and Compound A (CpdA), interfere with the NF-kappa B activation pathway in rheumatoid arthritis (RA) synovial cells. Methods. Quantitative polymerase chain reaction was performed to detect the tumor necrosis factor alpha (TNF alpha)-induced cytokine gene expression of interieukin-1 beta (IL-1 beta) and to investigate the effects of DEX and CpdA in RA fibroblast-like synoviocytes (FLS) transfected with small interfering RNA (siRNA) against GR (siGR) compared with nontransfected cells. Immunofluorescence analysis was used to detect the subcellular distribution of NF-kappa B (p65) under the various treatment conditions, and active DNA-bound p65 was measured using a TransAM assay and by chromatin immunoprecipitation analysis of IL-1 beta. Signaling pathways were studied via Western blotting of siGR-transfected cells, compared with nontransfected and nontargeting siRNA-transfected control cells, to detect the regulation of phospho-IKK, I kappa B alpha, phospho-p38, phospho-ERK, and phospho-JNK. Results. Both DEX and CpdA efficiently inhibited IL-1 beta gene expression in a GR-dependent manner. In addition, CpdA attenuated the TNF alpha-induced nuclear translocation and DNA binding of p65 in RA FLS, via the attenuation of IKK phosphorylation and subsequent I kappa B alpha degradation. CpdA also displayed profound effects on TNF alpha-induced MAPK activation. The effects of CpdA on TNF alpha-induced kinase activities occurred independently of the presence of GR. In sharp contrast, DEX did not affect TNF alpha-induced IKK phosphorylation, I kappa B alpha degradation, p65 nuclear translocation, or MAPK activation in RA FLS. Conclusion. DEX and CpdA display a dissimilar molecular mechanism of interaction with the NF-kappa B activation pathway ex vivo. A dual pathway, partially dependent and partially independent of GR (non-genomic), may explain the gene-inhibitory effects of CpdA in RA FLS.
Keywords
JOINT DESTRUCTION, INTERLEUKIN-6 GENE, MEDIATED REPRESSION, TRANSCRIPTIONAL ACTIVATION, COLLAGEN-INDUCED ARTHRITIS, TUMOR-NECROSIS-FACTOR, SYNOVIOCYTES, INFLAMMATORY CYTOKINE PRODUCTION, TRANSACTIVATION, PHOSPHORYLATION

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Chicago
Gossye, Valerie, Dirk Elewaut, Nadia Bougarne, Debby Bracke, Serge Van Calenbergh, Guy Haegeman, and Karolien De Bosscher. 2009. “Differential Mechanism of NF-κB Inhibition by Two Glucocorticoid Receptor Modulators in Rheumatoid Arthritis Synovial Fibroblasts.” Arthritis and Rheumatism 60 (11): 3241–3250.
APA
Gossye, V., Elewaut, D., Bougarne, N., Bracke, D., Van Calenbergh, S., Haegeman, G., & De Bosscher, K. (2009). Differential mechanism of NF-κB inhibition by two glucocorticoid receptor modulators in rheumatoid arthritis synovial fibroblasts. ARTHRITIS AND RHEUMATISM, 60(11), 3241–3250.
Vancouver
1.
Gossye V, Elewaut D, Bougarne N, Bracke D, Van Calenbergh S, Haegeman G, et al. Differential mechanism of NF-κB inhibition by two glucocorticoid receptor modulators in rheumatoid arthritis synovial fibroblasts. ARTHRITIS AND RHEUMATISM. 2009;60(11):3241–50.
MLA
Gossye, Valerie, Dirk Elewaut, Nadia Bougarne, et al. “Differential Mechanism of NF-κB Inhibition by Two Glucocorticoid Receptor Modulators in Rheumatoid Arthritis Synovial Fibroblasts.” ARTHRITIS AND RHEUMATISM 60.11 (2009): 3241–3250. Print.
@article{835808,
  abstract     = {Objective. To investigate and compare the molecular mechanisms by which 2 glucocorticoid receptor (GR)-activating compounds, dexamethasone (DEX) and Compound A (CpdA), interfere with the NF-kappa B activation pathway in rheumatoid arthritis (RA) synovial cells.
Methods. Quantitative polymerase chain reaction was performed to detect the tumor necrosis factor alpha (TNF alpha)-induced cytokine gene expression of interieukin-1 beta (IL-1 beta) and to investigate the effects of DEX and CpdA in RA fibroblast-like synoviocytes (FLS) transfected with small interfering RNA (siRNA) against GR (siGR) compared with nontransfected cells. Immunofluorescence analysis was used to detect the subcellular distribution of NF-kappa B (p65) under the various treatment conditions, and active DNA-bound p65 was measured using a TransAM assay and by chromatin immunoprecipitation analysis of IL-1 beta. Signaling pathways were studied via Western blotting of siGR-transfected cells, compared with nontransfected and nontargeting siRNA-transfected control cells, to detect the regulation of phospho-IKK, I kappa B alpha, phospho-p38, phospho-ERK, and phospho-JNK.
Results. Both DEX and CpdA efficiently inhibited IL-1 beta gene expression in a GR-dependent manner. In addition, CpdA attenuated the TNF alpha-induced nuclear translocation and DNA binding of p65 in RA FLS, via the attenuation of IKK phosphorylation and subsequent I kappa B alpha degradation. CpdA also displayed profound effects on TNF alpha-induced MAPK activation. The effects of CpdA on TNF alpha-induced kinase activities occurred independently of the presence of GR. In sharp contrast, DEX did not affect TNF alpha-induced IKK phosphorylation, I kappa B alpha degradation, p65 nuclear translocation, or MAPK activation in RA FLS.
Conclusion. DEX and CpdA display a dissimilar molecular mechanism of interaction with the NF-kappa B activation pathway ex vivo. A dual pathway, partially dependent and partially independent of GR (non-genomic), may explain the gene-inhibitory effects of CpdA in RA FLS.},
  author       = {Gossye, Valerie and Elewaut, Dirk and Bougarne, Nadia and Bracke, Debby and Van Calenbergh, Serge and Haegeman, Guy and De Bosscher, Karolien},
  issn         = {0004-3591},
  journal      = {ARTHRITIS AND RHEUMATISM},
  language     = {eng},
  number       = {11},
  pages        = {3241--3250},
  title        = {Differential mechanism of NF-\ensuremath{\kappa}B inhibition by two glucocorticoid receptor modulators in rheumatoid arthritis synovial fibroblasts},
  url          = {http://dx.doi.org/10.1002/art.24963},
  volume       = {60},
  year         = {2009},
}

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