title: Anti-inflammatory effects and the underlying mechanisms of action of daidzein in murine macrophages stimulated with Prevotella intermedia lipopolysaccharide.
abstract: OBJECTIVE
  Host modulatory agents directed at inhibiting specific proinflammatory mediators could be beneficial in terms of attenuating periodontal disease progression and potentially enhancing therapeutic responses. The aim of this study was to investigate whether daidzein could modulate the production inflammatory mediators in macrophages stimulated with lipopolysaccharide (LPS) from Prevotella intermedia, a pathogen implicated in periodontal disease, and to delineate underlying mechanisms of action.
  
  
  METHODS
  LPS was extracted from P. intermedia ATCC 25611 cells by the standard hot phenol-water method. The amounts of nitric oxide (NO) and interleukin-6 (IL-6) secreted into the culture medium were assayed. A real-time PCR was performed to quantify inducible nitric oxide synthase (iNOS) and IL-6 mRNA expression. We used immunoblot analysis to characterize iNOS protein expression, phosphrylation of c-Jun N-terminal kinase (JNK) and p38, degradation of inhibitory κB-α (IκB-α), nuclear translocation of nuclear factor-κB (NF-κB) subunits and phosphorylation of signal transducer and activator of transcription 1 (STAT1). The DNA-binding activity of NF-κB was assessed by using ELISA-based kits.
  
  
  RESULTS
  Daidzein significantly inhibited the production of NO and IL-6, as well as their mRNA expression, in P. intermedia LPS-treated RAW264.7 cells. The JNK and p38 pathways were not involved in the regulation of LPS-induced NO and IL-6 release by daidzein. Daidzein inhibited the degradation of IκB-α induced by P. intermedia LPS. In addition, daidzein suppressed NF-κB transcriptional activity via regulation of the nuclear translocation and DNA-binding activity of NF-κB p50 subunit and blocked STAT1 phosphorylation.
  
  
  CONCLUSIONS
  Although additional studies are required to dissect the molecular mechanism of action, our results suggest that daidzein could be a promising agent for treating inflammatory periodontal disease. Further research in animal models of periodontitis is necessary to better evaluate the potential of daidzein as a novel therapeutic agent to treat periodontal disease.
authors.affiliation: Department of Biological Science, College of Medical and Life Sciences, Silla University, Busan, Korea.
authors.forename: E-Y
authors.initials: EY
authors.lastname: Choi
authors.forename: J-Y
authors.initials: JY
authors.lastname: Jin
authors.forename: J-Y
authors.initials: JY
authors.lastname: Lee
authors.forename: J-I
authors.initials: JI
authors.lastname: Choi
authors.forename: I S
authors.initials: IS
authors.lastname: Choi
authors.forename: S-J
authors.initials: SJ
authors.lastname: Kim
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D001431:Bacteriological Techniques; D018929:Cell Culture Techniques; D002460:Cell Line; D006131:Growth Inhibitors / Q000494:pharmacology*; D051550:I-kappa B Kinase / Q000187:drug effects; D018836:Inflammation Mediators / Q000037:antagonists & inhibitors; D007529:Isoflavones / Q000494:pharmacology*; D048031:JNK Mitogen-Activated Protein Kinases / Q000187:drug effects; D053614:Janus Kinase 2 / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects*; D051379:Mice; D016328:NF-kappa B / Q000187:drug effects; D052002:NF-kappa B p50 Subunit / Q000187:drug effects; D052247:Nitric Oxide Synthase Type II / Q000187:drug effects; D010766:Phosphorylation; D048789:Phytoestrogens / Q000494:pharmacology*; D018713:Prevotella intermedia; D050794:STAT1 Transcription Factor / Q000187:drug effects; D051996:Transcription Factor RelA / Q000187:drug effects; D048051:p38 Mitogen-Activated Protein Kinases / Q000187:drug effects; D000093242:Interleukin-6 Inhibitors
chemical_list: D000893:Anti-Inflammatory Agents; D006131:Growth Inhibitors; D051550:I-kappa B Kinase; D018836:Inflammation Mediators; D007529:Isoflavones; D048031:JNK Mitogen-Activated Protein Kinases; D053614:Janus Kinase 2; D008070:Lipopolysaccharides; D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; D052247:Nitric Oxide Synthase Type II; D048789:Phytoestrogens; D050794:STAT1 Transcription Factor; D051996:Transcription Factor RelA; D048051:p38 Mitogen-Activated Protein Kinases; D000093242:Interleukin-6 Inhibitors; C494080:Stat1 protein, mouse; C004742:daidzein; C496317:Nos2 protein, mouse; C496564:Chuk protein, mouse