title: Euonymus alatus extract attenuates LPS-induced NF-κB activation via IKKβ inhibition in RAW 264.7 cells.
abstract: OBJECTIVE
  The present study was performed to investigate the underlying mechanisms of anti-inflammatory effects with the extract of Euonymus alatus (EEA), and specially focused on nuclear factor κB (NF-κB) signaling pathway by targeting the IκB kinase β (IKKβ).
  
  
  METHODS
  The effect of EEA for IKKβ activity was analyzed using an immobilized metal affinity for phosphochemicals (IMAP)-based time-resolved fluorescence resonance energy transfer (TR-FRET) assay. The effect of EEA on lipopolysaccharide (LPS)-induced NF-κB activation in murine macrophage RAW 264.7 cells with western blotting and immunofluorescent staining was evaluated.
  
  
  RESULTS
  IKKβ studies based on IMAP-TR-FRET showed that EEA possesses a potent IKKβ inhibitory activity with IC(50) value of 11.83μg/ml. EEA (10, 30μg/ml) also attenuated the LPS-induced IκBα phosphorylation/degradation, NF-κB translocation and subsequent NO synthesis in RAW 264.7 cells.
  
  
  CONCLUSIONS
  These results suggest that EEA abrogates LPS-induced NF-κB signaling pathway by targeting the IKKβ in RAW 264.7 cells and these properties may provide a molecular basis for understanding the inhibitory effects of EEA on LPS-mediated inflammation.
authors.affiliation: Bio-Organic Science Division, Korea Research Institute of Chemical Technology, 100, Jangdong, Yuseong, Daejeon 305-343, Republic of Korea.
authors.forename: Byung Koo
authors.initials: BK
authors.lastname: Oh
authors.forename: Jihye
authors.initials: J
authors.lastname: Mun
authors.forename: Ho Won
authors.initials: HW
authors.lastname: Seo
authors.forename: Shi Yong
authors.initials: SY
authors.lastname: Ryu
authors.forename: Young Sup
authors.initials: YS
authors.lastname: Kim
authors.forename: Byung Ho
authors.initials: BH
authors.lastname: Lee
authors.forename: Kwang-Seok
authors.initials: KS
authors.lastname: Oh
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology* / Q000627:therapeutic use; D001692:Biological Transport / Q000187:drug effects; D029756:Euonymus; D051550:I-kappa B Kinase / Q000037:antagonists & inhibitors*; D007249:Inflammation / Q000188:drug therapy; D020128:Inhibitory Concentration 50; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis; D010766:Phosphorylation; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology* / Q000627:therapeutic use; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D000893:Anti-Inflammatory Agents; D008070:Lipopolysaccharides; D016328:NF-kappa B; D010936:Plant Extracts; D009569:Nitric Oxide; D051550:I-kappa B Kinase