Anti-inflammatory effects and the underlying mechanisms of action of daidzein in murine macrophages stimulated with Prevotella intermedia lipopolysaccharide

• Published in: Journal of periodontal research Vol. 47; no. 2; pp. 204 - 211
• Main Authors: Choi, E.-Y., Jin, J.-Y., Lee, J.-Y., Choi, J.-I., Choi, I. S., Kim, S.-J.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01-04-2012; Blackwell
• Subjects: Animal models; Animals; Anti-Inflammatory Agents - pharmacology; Bacteriological Techniques; Biological and medical sciences; Bones, joints and connective tissue. Antiinflammatory agents; Cell Culture Techniques; Cell Line; daidzein; Dentistry; Facial bones, jaws, teeth, parodontium: diseases, semeiology; Growth Inhibitors - pharmacology; I-kappa B Kinase - drug effects; Inflammation Mediators - antagonists & inhibitors; interleukin-6; Interleukin-6 - antagonists & inhibitors; Isoflavones - pharmacology; Janus Kinase 2 - drug effects; JNK Mitogen-Activated Protein Kinases - drug effects; lipopolysaccharide; Lipopolysaccharides - pharmacology; Macrophages - drug effects; Medical sciences; Mice; NF-kappa B - drug effects; NF-kappa B p50 Subunit - drug effects; nitric oxide; Nitric Oxide Synthase Type II - drug effects; Non tumoral diseases; Otorhinolaryngology. Stomatology; p38 Mitogen-Activated Protein Kinases - drug effects; periodontal disease; Pharmacology. Drug treatments; Phosphorylation; Phytoestrogens - pharmacology; Prevotella intermedia; STAT1 Transcription Factor - drug effects; Transcription Factor RelA - drug effects; Stomatology; Rodentia; Antiinflammatory agent; interleukin-6; Bacteroidaceae; Interleukin 6; daidzein; Vertebrata; Periodontal disease; Mammalia; Prevotella intermedia; Mouse; Animal; Nitric oxide; Lipopolysaccharide; Bacteria; Mechanism of action; Macrophage
• ISSN: 0022-3484; 1600-0765
• DOI: 10.1111/j.1600-0765.2011.01422.x

Choi E‐Y, Jin J‐Y, Lee J‐Y, Choi J‐I, Choi IS, Kim S‐J. Anti‐inflammatory effects and the underlying mechanisms of action of daidzein in murine macrophages stimulated with Prevotella intermedia lipopolysaccharide. J Periodont Res 2012; 47: 204–211. © 2011 John Wiley & Sons A/S
Background and 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.
Material and 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.
Conclusion:  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.