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Suppression of LPS-induced inflammatory and NF-κB responses by anomalin in RAW 264.7 macrophages

✍ Scribed by Salman Khan; Eun Myoung Shin; Ran Joo Choi; Yoo Hyun Jung; Jinwoong Kim; Alev Tosun; Yeong Shik Kim

Publisher
John Wiley and Sons
Year
2011
Tongue
English
Weight
804 KB
Volume
112
Category
Article
ISSN
0730-2312

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✦ Synopsis

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-a (TNF-a) and interleukin-6 (IL-6), were reduced by anomalin, and this reduction correlated with the down-regulation of the NF-kB signaling pathway. In addition, anomalin suppressed the LPS-induced phosphorylation and degradation of IkBa. To further study the mechanisms underlying its anti-inflammatory activity, an electrophoretic mobility shift assay (EMSA) using a 32 P-labeled NF-kB probe was conducted. LPS-induced NF-kB 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.

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