Inhibition of LPS-induced TNF-alpha production by calcitonin gene-related peptide (CGRP) in cultured mouse peritoneal macrophages

Life Sci. 1997;61(20):PL 281-7. doi: 10.1016/s0024-3205(97)00866-7.

Abstract

The purpose of this study was to examine whether rCGRP has effects on TNF-alpha produced by mouse resident peritoneal macrophages. Macrophages were obtained from the peritoneal exudate of male Balb/c mouse. The cells were plated on culture dishes at a density of 2.5x10(5) cells per well and allowed to adhere for 2 hr. Pretreatment with rCGRP (10 nM-1 microM) for 24 hr, the macrophages were cultured with LPS 1 microg/ml for another 24 h. The medium was harvested for measuring TNF-alpha by ELISA kits. The results showed that rCGRP had no direct effects on TNF-alpha production, but it inhibited LPS-induced TNF-alpha production in a concentration-dependent manner. When rCGRP was at a concentration of 1 microM, the LPS-induced TNF-alpha production was inhibited by 39%. The effect of rCGRP was reversed by hCGRP(8-37) (10 microM), an antagonist of CGRP1 receptor. The LPS-induced TNF-alpha production from macrophages was also inhibited by forskolin 3 microM, an activator of adenylate cyclase. Furthermore, pretreatment with H-89 1 microM or Rp-cAMPS 100 microM, the inhibitors of cAMP-dependent protein kinase, the effect of rCGRP was abolished. These data suggest that the LPS-induced TNF-alpha production is inhibited by rCGRP via activation of cAMP responses in mouse resident peritoneal macrophages.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-Methyl-3-isobutylxanthine / pharmacology
  • Animals
  • Calcitonin Gene-Related Peptide / pharmacology*
  • Cells, Cultured
  • Cyclic AMP-Dependent Protein Kinases / antagonists & inhibitors
  • Enzyme Inhibitors / pharmacology
  • Isoquinolines / pharmacology
  • Lipopolysaccharides / pharmacology*
  • Macrophages, Peritoneal / metabolism*
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Recombinant Proteins / pharmacology
  • Sulfonamides*
  • Tumor Necrosis Factor-alpha / biosynthesis*

Substances

  • Enzyme Inhibitors
  • Isoquinolines
  • Lipopolysaccharides
  • Recombinant Proteins
  • Sulfonamides
  • Tumor Necrosis Factor-alpha
  • Cyclic AMP-Dependent Protein Kinases
  • Calcitonin Gene-Related Peptide
  • N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide
  • 1-Methyl-3-isobutylxanthine