Induction of cyclooxygenase-2 by platelet-derived growth factor (PDGF) and its inhibition by dexamethasone are independent of NF-kappaB/IkappaB transcription factors

Naunyn Schmiedebergs Arch Pharmacol. 2000 Jun;361(6):636-45. doi: 10.1007/s002100000231.

Abstract

Glucocorticoids are potent inhibitors of cyclooxygenase-2 (prostaglandin G/H synthase-2, COX-2) expression. The focus of this work was to investigate the molecular mechanisms, by which glucocorticoids interfere with platelet-derived growth factor (PDGF)-mediated induction of COX-2 with special emphasis on the role of the transcription factors NF-kappaB/IkappaB alpha. In rat renal mesangial cells, PDGF induced a rapid and transient increase of COX-2 mRNA and protein, which reached maximal levels after 1-2 and 4 h, respectively. The in vivo half-life of COX-2 mRNA, which was estimated to be less than 1 h, was reduced by dexamethasone. Kinetic studies and COX-2 promoter activity assays indicated that dexamethasone also interfered with COX-2 transcription. Inhibition of COX-2 induction by dexamethasone was abrogated by cycloheximide, an inhibitor of translation, indicating dependence on de novo protein synthesis. As a possible mediator of dexamethasone action, the NF-kappaB/IkappaB alpha system of transcription factors was investigated. Dexamethasone doubled IkappaB alpha protein levels within 1 h and reduced complex formation of nuclear NF-kappaB proteins with DNA. Newly synthesized IkappaB alpha may thus bind to NF-kappaB and interfere with gene activation. PDGF-induced signalling, however, barely affected the NF-kappaB/IkappaB alpha system: IkappaB alpha protein remained unaltered for 30 min after treatment of mesangial cells with PDGF and was only reduced by 30% after 1 h. Concomitantly, binding of NF-kappaB proteins to DNA, detected by electrophoretic mobility shift assays, was slightly increased by 30%. Furthermore, stably transfected COX-2 promoter constructs with and without the NF-KB binding site were comparably activated by PDGF (2.5-fold increase of luciferase activity). Taken together, these data indicate that although dexamethasone interferes with the NF-kappaB/IkappaB alpha system of transcription factors, this mechanism is not essential for the inhibition of PDGF-induced COX-2 expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Cycloheximide / pharmacology
  • Cyclooxygenase 2
  • DNA / metabolism
  • Dactinomycin / pharmacology
  • Dexamethasone / pharmacology*
  • Enzyme Induction
  • Gene Expression Regulation, Enzymologic
  • Genes, Reporter
  • I-kappa B Proteins / genetics
  • I-kappa B Proteins / metabolism*
  • Isoenzymes / biosynthesis*
  • Isoenzymes / genetics
  • Kidney Glomerulus / cytology
  • NF-kappa B / genetics
  • NF-kappa B / metabolism*
  • Nucleic Acid Synthesis Inhibitors / pharmacology
  • Platelet-Derived Growth Factor / pharmacology*
  • Prostaglandin-Endoperoxide Synthases / biosynthesis*
  • Prostaglandin-Endoperoxide Synthases / genetics
  • Protein Synthesis Inhibitors / pharmacology
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Transcriptional Activation

Substances

  • I-kappa B Proteins
  • Isoenzymes
  • NF-kappa B
  • Nucleic Acid Synthesis Inhibitors
  • Platelet-Derived Growth Factor
  • Protein Synthesis Inhibitors
  • RNA, Messenger
  • Dactinomycin
  • Dexamethasone
  • DNA
  • Cycloheximide
  • Cyclooxygenase 2
  • Prostaglandin-Endoperoxide Synthases