title: The effects of DA-6034 on NF-kappaB activity induced by lipopolysaccharide or tumor necrosis factor alpha in a human colonic epithelial cell line.
abstract: OBJECTIVE
  DA-6034, a synthetic derivative of eupatilin, has shown a potent anti-inflammatory effect in experimental rat models with inflammatory bowel disease (IBD). Nuclear factor kappaB (NF-kappaB) is known to play a central role in regulating inflammatory responses in patients with IBD, and is an important target for therapeutic intervention. We, therefore, examined the effects of DA-6034 on NF-kappaB activity and proinflammatory cytokines production in the human colonic cell line.
  
  
  METHODS
  HT-29 cells were stimulated with either lipopolysaccharide (LPS) or tumor necrosis factor alpha (TNF-alpha). After pretreatment with different doses of DA-6034, the influence of DA-6034 on NF-kappaB activity was determined by an electrophoretic mobility shift assay (EMSA). The effect of DA-6034 on inhibitory protein kappaBalpha (IkappaBalpha) activity was determined by immunoblotting. TNF-alpha and interlukin 8 (IL-8) messenger RNA expression, which are regulated by NF-kappaB, were assayed by RT-PCR. The production of TNF-alpha or IL-8 proteins was measured by ELISA.
  
  
  RESULTS
  DA-6034 inhibited NF-kappaB activity and prevented IkappaBalpha degradation in HT-29 cells after stimulation with either LPS or TNF-alpha. Furthermore, the production of TNF-alpha or IL-8 proteins decreased significantly after DA-6034 pretreatment.
  
  
  CONCLUSIONS
  These observations suggest that DA-6034 produces an anti-inflammaotory effect through inhibition of NF-kappaB activity by the prevention of I B degradation.
authors.affiliation: Department of Internal Medicine, Seoul Metropolitan Boramae Hospital, Seoul, Korea.
authors.forename: Ji Won
authors.initials: JW
authors.lastname: Kim
authors.forename: Byeong Gwan
authors.initials: BG
authors.lastname: Kim
authors.forename: Kook Lae
authors.initials: KL
authors.lastname: Lee
authors.forename: Ji Bong
authors.initials: JB
authors.lastname: Jeong
authors.forename: Yong Jin
authors.initials: YJ
authors.lastname: Jung
authors.forename: Joo Sung
authors.initials: JS
authors.lastname: Kim
authors.forename: Hyun Chae
authors.initials: HC
authors.lastname: Jung
authors.forename: In Sung
authors.initials: IS
authors.lastname: Song
mesh_terms: D002458:Cell Fractionation; D024202:Electrophoretic Mobility Shift Assay; D005419:Flavonoids / Q000008:administration & dosage / Q000494:pharmacology*; D019073:HT29 Cells; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D016328:NF-kappa B / Q000187:drug effects* / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000494:pharmacology
chemical_list: D005419:Flavonoids; D008070:Lipopolysaccharides; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; C121889:recoflavone