Inhibition of Helicobacter pylori-induced nuclear factor-kappa B activation and interleukin-8 gene expression by ecabet sodium in gastric epithelial cells

Helicobacter. 2003;8(5):542-53. doi: 10.1046/j.1523-5378.2003.00175.x.

Abstract

Background: Helicobacter pylori stimulates nuclear factor-kappa B (NF-kappa B) activation and chemokine interleukin-8 (IL-8) expression in gastric epithelial cells. Ecabet sodium (ecabet), a locally acting antiulcer drug, is known to have anti-H. pylori activity. However, there is little understanding of how ecabet induces anti-inflammatory activity in gastric epithelial cells infected with H. pylori. The aim of this study was to investigate the effects of ecabet on IL-8 gene expression and NF-kappa B activation in human gastric epithelial cells infected with H. pylori.

Materials and methods: After Hs746T, MKN-45, or SNU-5 gastric epithelial cell lines had been infected with cagA+cytotoxin+H. pylori in the presence of ecabet, IL-8 mRNA expression was assessed by quantitative reverse transcription-polymerase chain reaction, and IL-8 secretion was measured by enzyme-linked immunosorbent assay. NF-kappa B and inhibitory kappa B-alpha (I kappa B alpha) signals were assayed by electrophoretic mobility shift assay and Western blot, respectively. The activation of NF-kappa B and IL-8 reporter genes was determined by luciferase assay.

Results: Ecabet showed no antimicrobial activiy against Gram-positive or -negative bacteria. However, ecabet inhibited transcription of the IL-8 gene and secretion of IL-8 by gastric epithelial cells infected with H. pylori at a concentration of 5 micro g/ml. Moreover, ecabet inhibited the activation of NF- kappa B and the degradation of I kappa B alpha in gastric epithelial cells in response to H. pylori infection. In addition, the NF-kappa B signal inhibited by ecabet was comprised predominantly of heterodimers of p65/p50.

Conclusions: Ecabet inhibited H. pylori-induced IL-8 gene transcription and secretion by suppressing the NF-kappa B signal. This inhibition might be one pathway by which ecabet exerts its anti-inflammatory effect on H. pylori-induced gastric inflammation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abietanes / pharmacology*
  • Blotting, Western
  • Cell Line
  • Electrophoretic Mobility Shift Assay
  • Epithelial Cells / drug effects*
  • Epithelial Cells / metabolism
  • Epithelial Cells / microbiology
  • Gastric Mucosa / drug effects*
  • Gastric Mucosa / metabolism
  • Gastric Mucosa / microbiology
  • Gene Expression Regulation / drug effects
  • Genes, Reporter
  • Helicobacter pylori / drug effects
  • Helicobacter pylori / pathogenicity*
  • Humans
  • I-kappa B Proteins / metabolism
  • Interleukin-8 / biosynthesis*
  • Interleukin-8 / genetics
  • Luciferases / genetics
  • Luciferases / metabolism
  • Microbial Sensitivity Tests
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / metabolism*
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / isolation & purification
  • Transcription, Genetic / drug effects

Substances

  • Abietanes
  • I-kappa B Proteins
  • Interleukin-8
  • NF-kappa B
  • NFKBIA protein, human
  • RNA, Messenger
  • NF-KappaB Inhibitor alpha
  • ecabet
  • Luciferases