title: Anti-inflammatory activity of Ternstroemia gymnanthera stem bark extracts in bacterial lipopolysaccharide-stimulated RAW264.7 murine macrophage cells.
abstract: BACKGROUND
  Ternstroemia gymnanthera Sprague (Theaceae) possesses various known pharmacological properties. However, its anti-inflammatory activity has not been reported.
  
  
  OBJECTIVE
  The anti-inflammatory activity of Ternstroemia gymnanthera stem bark aqueous extract (TGSBE) was evaluated using LPS-stimulated RAW264.7 macrophages.
  
  
  METHODS
  Cytotoxicity was assessed by MTT assay after 24 h with TGSBE (25-200 μg/mL). Further testing used TGSBE at 100 and 200 μg/mL. Griess and ELISA methods after 24 h with TGSBE determined NO and cytokine levels, respectively; then, mRNA levels (iNOS & cytokines) were analyzed by Quantitative-PCR after 12 h. NF-κB and MAPK were assessed by immunoblotting after TGSBE treatment for 12 h, followed by LPS for 30 min. Immunofluorescence assay was also performed for NF-κB. ROS and MMP, after 12 h with TGSBE, were determined by flow cytometry. The antioxidant potential of TGSBE was analyzed by ABTS assay. The Folin-Ciocalteu method determined the total phenolic content of TGSBE. LPS concentration was 0.5 μg/mL.
  
  
  RESULTS
  TGSBE at 200 μg/mL showed about 96.2% viability while suppressing the production of NO (88.99%), TNFα (24.38%), IL-6 (61.70%) and IL-1β (55.12%) and gene expression by 67.88, 45.24, 65.84, and 70.48%, respectively. TGSBE decreased ROS (79.26%) and improved MMP (48.01%); it inhibited translocation of NF-κB and MAPK activation. Radical scavenging activity was 50% at 402.17 μg/mL (ascorbic acid standard: 88.8 μg/mL). Total phenolic content was 240.9 mg GAE/g.
  
  
  CONCLUSIONS
  TGSBE suppresses the inflammatory response by inhibiting the NF-κB and MAPK cascades exhibiting therapeutic potential to treat inflammatory diseases associated with increased activation of macrophages.
authors.affiliation: a Department of Forest products and Biotechnology , College of Forest Science, Kookmin University , Seoul , South Korea.
authors.forename: Thamizhiniyan
authors.initials: T
authors.lastname: Venkatesan
authors.affiliation: a Department of Forest products and Biotechnology , College of Forest Science, Kookmin University , Seoul , South Korea.
authors.forename: Eun-Jin
authors.initials: EJ
authors.lastname: Park
authors.affiliation: a Department of Forest products and Biotechnology , College of Forest Science, Kookmin University , Seoul , South Korea.
authors.forename: Young-Woong
authors.initials: YW
authors.lastname: Choi
authors.affiliation: a Department of Forest products and Biotechnology , College of Forest Science, Kookmin University , Seoul , South Korea.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Lee
authors.affiliation: a Department of Forest products and Biotechnology , College of Forest Science, Kookmin University , Seoul , South Korea.
authors.forename: Young-Kyoon
authors.initials: YK
authors.lastname: Kim
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000494:pharmacology*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis; D010936:Plant Extracts / Q000494:pharmacology*; D018547:Plant Stems; D000067996:RAW 264.7 Cells; D017382:Reactive Oxygen Species / Q000378:metabolism; D028242:Theaceae
chemical_list: D000893:Anti-Inflammatory Agents; D016207:Cytokines; D008070:Lipopolysaccharides; D016328:NF-kappa B; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D009569:Nitric Oxide
keywords: Ornamental plant; inflammation; natural antioxidants; traditional medicine