title: DA-9601, a standardized extract of Artemisia asiatica, blocks TNF-alpha-induced IL-8 and CCL20 production by inhibiting p38 kinase and NF-kappaB pathways in human gastric epithelial cells.
abstract: OBJECTIVE
  To investigate whether, or how, DA-9601, which is a new gastroprotective agent, inhibits TNF-alpha-induced inflammatory signals in gastric epithelial AGS cells.
  
  
  METHODS
  Cell viability was determined by MTT assay. IL-8 and CCL20 promoter activities were determined by a luciferase reporter gene assay. NF-kappaB-dependent transcriptional activity was determined by I-kappaB alpha degradation, NF-kappaB p65 nuclear translocation and a luciferase activity assay. IL-8 and CCL20 gene expression and protein secretion were determined by RT-PCR and an enzyme-linked immunosorbent assay (ELISA). Total and phosphorylated forms of mitogen-activated protein kinases (MAPKs) were determined by Western blot.
  
  
  RESULTS
  Treatment of AGS cells with DA-9601 reduced TNF-alpha-induced IL-8 and CCL20 promoter activities, as well as their gene expression and protein release. TNF-alpha also induced NF-kappaB-dependent transcriptional activity in AGS cells. In contrast, in cells treated with DA-9601, TNF-alpha-induced NF-kappaB activity was significantly blocked. Although all three MAP kinase family members were phosphorylated in response to TNF-alpha, a selective inhibitor of p38 kinase SB203580 only could inhibit both NF-kappaB-dependent transcriptional activity and IL-8 and CCL20 production, suggesting a potential link between p38 kinase and NF-kappaB-dependent pathways in AGS cells. Interestingly, DA-9601 also selectively inhibited p38 kinase phosphorylation induced by TNF-alpha.
  
  
  CONCLUSIONS
  DA-9601 blocked TNF-alpha-mediated inflammatory signals by potentially modulating the p38 kinase pathway and/or a signal leading to NF-kappaB-dependent pathways in gastric epithelial cells.
authors.affiliation: Digestive Disease Research Institute, Wonkwang University School of Medicine, Chonbuk, Korea.
authors.forename: Suck-Chei
authors.initials: SC
authors.lastname: Choi
authors.forename: Eun-Ju
authors.initials: EJ
authors.lastname: Choi
authors.forename: Hyun-Mee
authors.initials: HM
authors.lastname: Oh
authors.forename: SungGa
authors.initials: S
authors.lastname: Lee
authors.forename: Jeong-Kun
authors.initials: JK
authors.lastname: Lee
authors.forename: Meung-Su
authors.initials: MS
authors.lastname: Lee
authors.forename: Yong-Il
authors.initials: YI
authors.lastname: Shin
authors.forename: Suck-Jun
authors.initials: SJ
authors.lastname: Choi
authors.forename: Jeong-Ryong
authors.initials: JR
authors.lastname: Chae
authors.forename: Kang-Min
authors.initials: KM
authors.lastname: Lee
authors.forename: Won-Jung
authors.initials: WJ
authors.lastname: Lee
authors.forename: Jae-Sik
authors.initials: JS
authors.lastname: Park
authors.forename: Chang-Yell
authors.initials: CY
authors.lastname: Shin
authors.forename: Tae-Young
authors.initials: TY
authors.lastname: Oh
authors.forename: Chang-Duk
authors.initials: CD
authors.lastname: Jun
mesh_terms: D019306:Artemisia / Q000737:chemistry; D054418:Chemokine CCL20; D019742:Chemokines, CC / Q000235:genetics / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005753:Gastric Mucosa / Q000166:cytology; D017930:Genes, Reporter; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism*; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D019402:Macrophage Inflammatory Proteins / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D015398:Signal Transduction / Q000502:physiology; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: C108355:CCL20 protein, human; D054418:Chemokine CCL20; D019742:Chemokines, CC; C116959:DA 9601; D004791:Enzyme Inhibitors; D016209:Interleukin-8; D019402:Macrophage Inflammatory Proteins; D016328:NF-kappa B; D010936:Plant Extracts; D014409:Tumor Necrosis Factor-alpha; D048049:Extracellular Signal-Regulated MAP Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases