title: [Effect of genistein on the TLR and MAPK transduction cascades in lipopolysaccharide -stimulated macrophages].
abstract: OBJECTIVE
  To delineate and confirm the signaling processes involving mitogen-activated protein kinase (MAPK) and Toll-like receptor (TLR) in the regulation of lipopolysaccharide (LPS)-induced activity of macrophages by genistein at the protein and transcriptional levels.
  
  
  METHODS
  RAW264.7 macrophages were treated with genistein and then stimulated with LPS (100 ng/mL) for different time duration. We evaluated the induction of MAPK phosphorylation by Western blot analysis; RT2 Profiler(TM); PCR array was used to investigate the expressions of TLR pathway-related genes after different treatments.
  
  
  RESULTS
  LPS led to the phosphorylation of Raf, MEK1/2 and ERK1/2 at 15 minutes post-stimulation and it lasted till 30 minutes. Phosphorylation of p38 was also observed, but it was not as obvious as p-ERK1/2. Addition of genistein further increased the phosphorylation of ERK1/2, its downstream protein p90rsk and p38. Cells treated with LPS and LPS together with genistein demonstrated significant number of genes to be differentially regulated as compared with control cells. Genistein alone could up-regulate the gene CD80, MEKK1, c-fos, Rela, and Ticam2. LPS could up-regulate 20 genes including cytokines IFN-β, IL-10, IL-1α, IL-1β, IL-6, TNF-α, colony stimulating factor 2 (CSF-2) and CSF-3, chemokines CCL2 and CXCL10, transcription factor NF-κB1, IκB-α and cyclooxygenase 2 (COX-2). The presence of genistein led to a strong inhibition of the expressions of these genes and up-regulated the transcription factors IκB-β and c-Rel, a subunit of NF-κB.
  
  
  CONCLUSIONS
  Genistein strongly enhances the LPS-induced activities of MAPK transduction cascades and inhibits TLR pathway.
authors.affiliation: Medical College, Yangzhou University, Yangzhou 225001, China; Helmholtz Center for Infection Research, Braunschweig 38124, Germany.
authors.forename: Shuna
authors.initials: S
authors.lastname: Cui
authors.affiliation: Helmholtz Center for Infection Research, Braunschweig 38124, Germany.
authors.forename: Ursula
authors.initials: U
authors.lastname: Bilitewski
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D019833:Genistein / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D015398:Signal Transduction / Q000187:drug effects*; D051193:Toll-Like Receptors / Q000502:physiology*
chemical_list: D016207:Cytokines; D008070:Lipopolysaccharides; D051193:Toll-Like Receptors; D019833:Genistein; D048049:Extracellular Signal-Regulated MAP Kinases