Interleukin 1 beta-induced expression of nitric oxide synthase in rat renal mesangial cells is suppressed by cyclosporin A

Biochem Biophys Res Commun. 1995 Nov 13;216(2):438-46. doi: 10.1006/bbrc.1995.2642.

Abstract

The expression of inducible nitric oxide synthase (iNOS) is triggered in rat renal mesangial cells by exposure to the inflammatory cytokine interleukin 1 beta (IL-1 beta). Here we report that cyclosporin A (CsA) a potent immunosuppressive drug, inhibits IL-1 beta dependent iNOS expression in renal mesangial cells. Addition of CsA dose dependently suppresses IL-1 beta-induced nitrite formation (IC50 = 0.9 microM). Western- and Northern blot analyses of mesangial cell extracts reveal that the inhibition of IL-1 beta-induced nitrite formation by CsA is due to decreased iNOS protein and iNOS mRNA steady state levels. Using nuclear run on experiments we show that the transcription rate of the IL-1 beta-induced iNOS gene is reduced. Furthermore, by electrophoretic mobility shift analysis we demonstrate reduced DNA-binding of the nuclear factor NF kappa B, an essential component of the IL-1 beta-dependent upregulation of iNOS gene transcription. The data presented in this report suggest that the cellular machinery involved in the IL-1 beta dependent transcriptional upregulation of the iNOS gene in mesangial cells is a target for the action of CsA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Northern
  • Blotting, Western
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • Cyclosporine / pharmacology*
  • Enzyme Induction
  • Gene Expression / drug effects*
  • Glomerular Mesangium / enzymology*
  • Humans
  • Interleukin-1 / antagonists & inhibitors
  • Interleukin-1 / pharmacology*
  • Kinetics
  • NF-kappa B / drug effects
  • NF-kappa B / metabolism
  • Nitric Oxide Synthase / analysis
  • Nitric Oxide Synthase / biosynthesis*
  • RNA, Messenger / analysis
  • RNA, Messenger / biosynthesis
  • Rats
  • Recombinant Proteins / pharmacology
  • Transcription, Genetic / drug effects

Substances

  • Interleukin-1
  • NF-kappa B
  • RNA, Messenger
  • Recombinant Proteins
  • Cyclosporine
  • Nitric Oxide Synthase