Endotoxin, but not platelet-activating factor, activates nuclear factor-κB and increases IκBα and IκBβ turnover in enterocytes

Isabelle G. De Plaen, Xiao Wu Qu, Hao Wang, Xiao Di Tan, Liya Wang, Xin Bing Han, Ranna A. Rozenfeld, Wei Hsueh*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

Bacterial endotoxin (lipopolysaccharide; LPS) and platelet-activating factor (PAF) are important triggers of bowel inflammation and injury. We have previously shown that LPS activates the transcription factor nuclear factor (NF)-κB in the intestine, which up-regulates many pro-inflammatory genes. This effect partly depends on neutrophils and endogenous PAF. However, whether LPS and PAF directly activate NF-κB in enterocytes remains controversial. In this study, we first investigated the effect of LPS and PAF on NF-κB activation in IEC-6 (a non-transformed rat small intestinal crypt cell line) cells, by electrophoresis mobility shift assay and supershift, and found that LPS, but not PAF, activates NF-κB mostly as p50-p65 heterodimers. The effect was slower than tumour necrosis factor (TNF). Both LPS and TNF induce the expression of the NF-κB-dependent gene inducible nitric oxide synthase (iNOS), which occurs subsequent to NF-κB activation. We then examined the effect of LPS and TNF on the inhibitory molecules IκBα and IκBβ. We found that TNF causes rapid degradation of IκBβ and IκBβ. In contrast, LPS did not change the levels of IκBα and IκBβ up to 4 hr (by Western blot). However, in the presence of cycloheximide, there was a slow reduction of IκBα and IκBβ, which disappeared almost completely at 4 hr. These observations suggest that LPS causes slow degradation and synthesis of IκBα and IκBβ and therefore activates NF-κB via at least two mechanisms: initially, through an IκB-independent mechanism, and later, via an increased turnover of the inhibitor IκB. NF-κB activation precedes the gene expression of iNOS (assayed by reverse transcription-polymerase chain reaction), suggesting that LPS up-regulates iNOS via this transcription factor.

Original languageEnglish (US)
Pages (from-to)577-583
Number of pages7
JournalImmunology
Volume106
Issue number4
DOIs
StatePublished - 2002

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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