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Dexamethasone regulates AP-1 to repress TNF-alpha induced MCP-1 production in human glomerular endothelial cells
Su-Kil Park1, Won Seok Yang, Nam Jung Han
1Department of Internal Medicine, Asan Medical Center, College of Medicine, University of Ulsan, Seoul, Korea. skpark@www.amc.seoul.kr
Background:
Glomerular endothelial cells play a role in the pathogenesis of glomerulonephritis by producing chemotactic factors. We investigated the role of NF-kappa B and AP-1 in tumour necrosis factor alpha (TNF-alpha) induced monocyte chemoattractant protein 1 (MCP-1) production in cultured human glomerular endothelial cells (HGEC). We also examined whether or not these processes could be modified by glucocorticoid.
Methods:
MCP-1 protein and mRNA levels were measured by ELISA and northern blot. NF-kappa B and AP-1 binding activity were assessed by electrophoretic mobility shift assay. Cytosolic I kappa B alpha and nuclear p65 protein were evaluated by western blot. For specific inhibition of NF-kappa B or AP-1, we used a decoy oligodeoxynucleotide.
Results:
TNF-alpha (10 ng/ml) increased MCP-1 mRNA expression in HGEC and also the release of MCP-1 protein into culture media. These effects could be partially inhibited by dexamethasone (10 nM). TNF-alpha induced MCP-1 production appeared to be NF-kappa B and AP-1 interdependent, based on the following results. (i) TNF-alpha increased NF-kappa B and AP-1 binding activity. (ii) Both NF-kappa B decoy oligodeoxynucleotide and AP-1 decoy oligodeoxynucleotide partially suppressed TNF-alpha induced MCP-1 mRNA expression. On the other hand, dexamethasone decreased TNF-alpha induced DNA-binding activity of AP-1 without an effect on the DNA-binding activity of NF-kappa B, cytosolic I kappa B alpha degradation or p65 nuclear translocation.
Conclusions:
These data demonstrate that while TNF-alpha induced MCP-1 production is mediated by the cooperative action of NF-kappa B and AP-1 in HGEC, dexamethasone represses TNF-alpha induced MCP-1 production via suppression of AP-1 binding activity.
