Premium
Suppression of LPS‐induced inflammatory and NF‐κB responses by anomalin in RAW 264.7 macrophages
Author(s) -
Khan Salman,
Shin Eun Myoung,
Choi Ran Joo,
Jung Yoo Hyun,
Kim Jinwoong,
Tosun Alev,
Kim Yeong Shik
Publication year - 2011
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.23137
Subject(s) - tumor necrosis factor alpha , nf κb , nitric oxide , nitric oxide synthase , macrophage , inflammation , proinflammatory cytokine , chemistry , electrophoretic mobility shift assay , signal transduction , mechanism of action , pharmacology , phosphorylation , microbiology and biotechnology , biochemistry , immunology , biology , gene expression , in vitro , gene , organic chemistry
The treatment of inflammatory diseases today is largely based on interrupting the synthesis or action of the mediators that drive the host's response to injury. It is on the basis of this concept that most of the anti‐inflammatory drugs have been developed. In our continuous search for novel anti‐inflammatory agents from traditional medicinal plants, Saposhnikovia divaricata has been a focus of our investigations. Anomalin, a pyranocoumarin constituent of S. divaricata , exhibits potent anti‐inflammatory activity. To clarify the cellular signaling mechanisms underlying the anti‐inflammatory action of anomalin, we investigated the effect of anomalin on the production of inflammatory molecules in LPS‐stimulated murine macrophages. The anomalin dose‐dependently inhibited inducible nitric oxide synthase (iNOS) and cyclooxygenase‐2 (COX‐2) mRNA and protein expression in LPS‐stimulated RAW 264.7 macrophage. Molecular analysis using quantitative real time polymerase chain reaction (qRT‐PCR) revealed that several pro‐inflammatory cytokines, including tumor necrosis factor‐α (TNF‐α) and interleukin‐6 (IL‐6), were reduced by anomalin, and this reduction correlated with the down‐regulation of the NF‐κB signaling pathway. In addition, anomalin suppressed the LPS‐induced phosphorylation and degradation of IκBα. To further study the mechanisms underlying its anti‐inflammatory activity, an electrophoretic mobility shift assay (EMSA) using a 32 P‐labeled NF‐κB probe was conducted. LPS‐induced NF‐κB DNA binding was drastically abolished by anomalin. The present data suggest that anomalin is a major anti‐inflammatory agent and may be a potential therapeutic candidate for the treatment of inflammatory disorders. J. Cell. Biochem. 112: 2179–2188, 2011. © 2011 Wiley‐Liss, Inc.