Short-term treatment of RAW264.7 macrophages with adiponectin increases tumor necrosis factor-alpha (TNF-alpha) expression via ERK1/2 activation and Egr-1 expression: role of TNF-alpha in adiponectin-stimulated interleukin-10 production

J Biol Chem. 2007 Jul 27;282(30):21695-703. doi: 10.1074/jbc.M701419200. Epub 2007 May 30.

Abstract

Adiponectin is an adipokine with potent anti-inflammatory properties. However, the mechanisms by which adiponectin suppresses macrophage function are not well understood. Treatment of RAW264.7 macrophages with adiponectin for 18 h decreased lipopolysaccharide (LPS)-stimulated tumor necrosis factor-alpha (TNF-alpha) production. Here we demonstrate that globular adiponectin (gAcrp) initially increased TNF-alpha expression in RAW264.7 macrophages; this TNF-alpha then contributed to increased expression of interleukin-10, which in turn was required for the development of tolerance to subsequent LPS exposure. gAcrp-mediated increases in TNF-alpha mRNA accumulation were associated with increased TNF-alpha promoter activity. gAcrp increased the DNA binding activity of both Egr-1 and NFkappaB; mutation of either the Egr-1 or NFkappaB binding sites in the TNF-alpha promoter decreased gAcrp-stimulated promoter activity. Further, co-transfection with either dominant negative Egr-1 or the IkappaB super-repressor prevented gAcrp-stimulated TNF-alpha promoter activity. gAcrp also increased Egr-1 promoter activity, mRNA accumulation, and DNA binding activity. Inhibition of ERK1/2 with U0126 potently suppressed gAcrp-stimulated Egr-1 promoter activity, as well as TNF-alpha promoter activity. In summary, these data demonstrate that adiponectin initially increases TNF-alpha production by macrophages via ERK1/2-->Egr-1 and NFkappaB-dependent mechanisms; these increases in TNF-alpha in turn lead to increased expression of interleukin-10 and an eventual dampening of LPS-mediated cytokine production in macrophages.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adiponectin / pharmacology*
  • Animals
  • Cell Line
  • Cytokines / biosynthesis
  • DNA Primers
  • Early Growth Response Protein 1 / genetics*
  • Enzyme Activation
  • Genes, Reporter
  • Interleukin-10 / biosynthesis*
  • Lipopolysaccharides / pharmacology
  • Luciferases / genetics
  • Macrophages / drug effects
  • Macrophages / enzymology
  • Macrophages / physiology*
  • Mice
  • Mitogen-Activated Protein Kinase 1 / metabolism*
  • Mitogen-Activated Protein Kinase 3 / metabolism*
  • NF-kappa B / metabolism
  • Phosphorylation
  • Tumor Necrosis Factor-alpha / genetics*

Substances

  • Adiponectin
  • Cytokines
  • DNA Primers
  • Early Growth Response Protein 1
  • Egr1 protein, mouse
  • Lipopolysaccharides
  • NF-kappa B
  • Tumor Necrosis Factor-alpha
  • Interleukin-10
  • Luciferases
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3