Selective inhibition of inhibitory kappa B kinase-β abrogates induction of nitric oxide synthase in lipopolysaccharide-stimulated rat aortic smooth muscle cells

Almudena Bermejo Gomez, Christopher MacKenzie, Andrew Paul, Robin Plevin

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13 Citations (Scopus)

Abstract

In this study, we utilised a number of adenoviral constructs in order to examine the role of intermediates of the NF-κB pathway in the regulation of inducible nitric oxide synthase (iNOS) induction in rat aortic smooth muscle cells (RASMCs). Lipopolysaccharide (LPS) stimulated a significant increase in iNOS induction and NF-κB DNA binding. These parameters were substantially reduced by overexpression of a wild-type Iκ-Bα adenoviral construct (Ad.Iκ-Bα), confirming a role for NF-κB in iNOS induction. Infection with a dominant-negative IKKα adenoviral construct (Ad.IKKα +/-) did not significantly affect iNOS induction, NF-κB DNA binding or Iκ-Bα loss. Infection of RASMCs with adenovirus encoding a dominant-negative IKKβ (Ad.IKKβ +/-) essentially abolished iNOS induction and activation of the NF-κB pathway. Pretreatment of RASMCs with a novel specific inhibitor of IKKβ, SC-514, significantly reduced iNOS induction, NF-κB DNA binding and I-κBα loss in a concentration-dependent manner. In both RASMCs and human umbilical vein endothelial cells (HUVECs), infection with Ad.IKKα +/- also inhibited COX-2 expression in response to LPS. However, Ad.IKKα +/- was again without effect. These data suggest that IKKβ plays a predominant, selective role in the regulation of NF-κB-dependent induction of iNOS in RASMCs.

Original languageEnglish
Pages (from-to)217-225
Number of pages9
JournalBritish Journal of Pharmacology
Volume146
Issue number2
DOIs
Publication statusPublished - 1 Dec 2005

Keywords

  • aortic smooth muscle cells
  • inducible nitric oxide synthase
  • inhibitory kappa B kinase
  • lipopolysaccharide
  • nuclear factor kappa B

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