8β-hydroxy-3-oxopimar-15-ene exerts anti-inflammatory effects by inhibiting ROS-mediated activation of the TRAF6-ASK1-p38 signaling pathway

Immunopharmacol Immunotoxicol. 2013 Oct;35(5):549-57. doi: 10.3109/08923973.2013.820742. Epub 2013 Aug 5.

Abstract

The flying squirrel's droppings (Pteropus pselaphon) have been used for improving the blood circulation, arresting bleeding to treat hematological disorders, and reducing pain. Here, 8β-hydroxy-3-oxopimar-15-ene (OXO), one of main constituents of P. pselaphon, was examined for its anti-inflammatory activity in murine macrophages. We found that OXO significantly suppressed LPS-induced nitric oxide (NO) without exerting cytotoxic effects on RAW 264.7 cells. OXO inhibited the expression of LPS-induced iNOS and COX-2 protein and their mRNA in a dose-dependent manner. Also, TNF-α, IL-6, and PGE2 secretion was decreased by OXO in LPS-stimulated macrophages. These inflammatory biomarkers were attributed to the suppression of LPS-induced activation of p38 MAPK and subsequent activation of two components of AP-1 (c-Jun and c-Fos), but not of ERK, JNK, NF-κB. Moreover, OXO inhibited LPS-induced intracellular reactive oxygen species (ROS) production and co-incubation of OXO and hydrogen peroxide (H2O2) suppressed the phosphorylation of p38 in a concentration-dependent manner. In addition, OXO completely disrupted the formation of TRAF6-ASK complex in the cells. Therefore, we demonstrate here that OXO can potentially inhibit several biomarkers related to inflammation through inhibition of ROS-mediated activation of TRAF6-ASK1-p38 pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / pharmacology*
  • Cell Line
  • Cyclooxygenase 2 / immunology
  • Cyclooxygenase 2 / metabolism
  • Dinoprostone / immunology
  • Dinoprostone / metabolism
  • Diterpenes / chemistry
  • Diterpenes / pharmacology*
  • Dose-Response Relationship, Drug
  • Interleukin-6 / immunology
  • Interleukin-6 / metabolism
  • Lipopolysaccharides / toxicity
  • MAP Kinase Kinase Kinase 5 / immunology*
  • MAP Kinase Kinase Kinase 5 / metabolism
  • MAP Kinase Signaling System / drug effects*
  • MAP Kinase Signaling System / immunology
  • Macrophages
  • Mice
  • Nitric Oxide / immunology
  • Nitric Oxide / metabolism
  • Nitric Oxide Synthase Type II / immunology
  • Nitric Oxide Synthase Type II / metabolism
  • RNA, Messenger / immunology
  • RNA, Messenger / metabolism
  • Reactive Oxygen Species / immunology*
  • Reactive Oxygen Species / metabolism
  • Sciuridae
  • TNF Receptor-Associated Factor 6 / immunology*
  • TNF Receptor-Associated Factor 6 / metabolism
  • Tumor Necrosis Factor-alpha / immunology
  • Tumor Necrosis Factor-alpha / metabolism
  • p38 Mitogen-Activated Protein Kinases / immunology*
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • 8-hydroxy-3-oxopimar-15-ene
  • Anti-Inflammatory Agents
  • Diterpenes
  • Interleukin-6
  • Lipopolysaccharides
  • RNA, Messenger
  • Reactive Oxygen Species
  • TNF Receptor-Associated Factor 6
  • Tumor Necrosis Factor-alpha
  • Nitric Oxide
  • Nitric Oxide Synthase Type II
  • Nos2 protein, mouse
  • Ptgs2 protein, mouse
  • Cyclooxygenase 2
  • p38 Mitogen-Activated Protein Kinases
  • MAP Kinase Kinase Kinase 5
  • Map3k5 protein, mouse
  • Dinoprostone