Genistein suppresses Prevotella intermedia lipopolysaccharide-induced inflammatory response in macrophages and attenuates alveolar bone loss in ligature-induced periodontitis

Arch Oral Biol. 2016 Feb:62:70-9. doi: 10.1016/j.archoralbio.2015.11.019. Epub 2015 Nov 26.

Abstract

Objective: Genistein is a major isoflavone subclass of flavonoids found in soybean and a potent tyrosine kinase inhibitor. The present study aimed to assess the effect of genistein on the production of proinflammatory mediators in murine macrophages stimulated with lipopolysaccharide (LPS) isolated from Prevotella intermedia, a pathogen associated with different forms of periodontal disease, and to evaluate its possible influence on alveolar bone loss in ligature-induced periodontitis using micro-computed tomography (micro-CT) analysis as well.

Design: LPS was isolated from P. intermedia ATCC 25611 by using the standard hot phenol-water method. Culture supernatants were analyzed for nitric oxide (NO) and interleukin-6 (IL-6). Inducible NO synthase (iNOS) protein expression was evaluated by immunoblot analysis. Real-time PCR was carried out to measure iNOS and IL-6 mRNA expression. In addition, effect of genistein on alveolar bone loss was evaluated in a rat model of experimental periodontitis using micro-CT analysis.

Results: Genistein significantly attenuated P. intermedia LPS-induced production of iNOS-derived NO and IL-6 with attendant decrease in their mRNA expression in RAW264.7 cells. In addition, when genistein was administered to rats, decreases in alveolar bone height and bone volume fraction induced by ligature placement were significantly inhibited. Genistein administration also prevented ligature-induced alterations in the microstructural parameters of trabecular bone, including trabecular thickness, trabecular separation, bone mineral density and structure model index.

Conclusions: While additional studies are required, we suggest that genistein could be utilized for the therapy of human periodontitis in the future.

Keywords: Experimental periodontitis; Genistein; Lipopolysaccharide; Micro-computed tomography; Prevotella intermedia; Proinflammatory mediators.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alveolar Bone Loss / drug therapy*
  • Alveolar Bone Loss / genetics
  • Alveolar Bone Loss / metabolism
  • Animals
  • Disease Models, Animal
  • Gene Expression
  • Genistein / pharmacology*
  • Inflammation Mediators / antagonists & inhibitors*
  • Interleukin-6 / antagonists & inhibitors
  • Interleukin-6 / biosynthesis
  • Ligation
  • Lipopolysaccharides / antagonists & inhibitors*
  • Lipopolysaccharides / isolation & purification
  • Lipopolysaccharides / pharmacology
  • Macrophages / drug effects*
  • Macrophages / immunology
  • Male
  • Mice
  • Nitric Oxide / biosynthesis
  • Nitric Oxide Synthase Type II / antagonists & inhibitors
  • Nitric Oxide Synthase Type II / biosynthesis
  • Periodontitis / drug therapy*
  • Periodontitis / genetics
  • Periodontitis / immunology
  • Periodontitis / metabolism
  • Prevotella intermedia / chemistry
  • RAW 264.7 Cells
  • RNA, Messenger / genetics
  • Random Allocation
  • Rats
  • Rats, Sprague-Dawley

Substances

  • IL6 protein, human
  • Inflammation Mediators
  • Interleukin-6
  • Lipopolysaccharides
  • RNA, Messenger
  • Nitric Oxide
  • Genistein
  • NOS2 protein, human
  • Nitric Oxide Synthase Type II