Src/Syk/IRAK1-targeted anti-inflammatory action of Torreya nucifera butanol fraction in lipopolysaccharide-activated RAW264.7 cells. (21st July 2016)

• Record Type: Journal Article
• Authors: Kim, Shi Hyoung; Park, Jae Gwang; Hong, Yong Deog; Kim, Eunji; Baik, Kwang-Soo; Yoon, Deok Hyo; Kim, Sunggyu; Lee, Mi-nam; Rho, Ho Sik; Shin, Song Seok; Cho, Jae Youl
• Is Part Of: Journal of ethnopharmacology. Volume 188(2016)
• Page Start: 167
• Page End: 176
• Publication Date: 2016-07-21
• Subjects: PG prostaglandin -- NO nitric oxide -- COX cyclooxygenase -- iNOS inducible NO synthase -- TNF-α tumor necrosis factor α -- ERK extracellular signal-related kinase -- TLR Toll-like receptors -- MAPK mitogen activated protein kinase -- NF-κB nuclear factor-κB -- AP-1 activator protein-1 -- JNK c-Jun N-terminal kinase -- EIA enzyme immunoassay -- ELISA enzyme linked immunosorbent assay -- MTT 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (a tetrazole) -- FRA-1 Fos-related antigen 1 -- PI3K phosphoinositide 3-kinase -- LPS lipopolysaccharide -- and RT-PCR reverse transcriptase-polymerase chain reaction; Torreya nucifera -- Taxaceae -- Anti-inflammatory activities -- NF-κB -- AP-1 -- Src -- IRAK1 -- Syk; Ethnopharmacology -- Periodicals; Pharmacognosy -- Periodicals; Herbs -- Periodicals; Herbs -- Periodicals; Pharmacognosy -- Periodicals; Pharmacognosie -- Périodiques; Herbes -- Périodiques; 615.1
• DOI: 10.1016/j.jep.2016.05.008
• Languages: English
• ISSNs: 0378-8741

Abstract

Abstract: Ethnopharmacological relevance: Seed of Torreya nucifera (L.) Siebold & Zucc is used to treat several diseases in Asia. Reports document that T. nucifera has anti-cancer, anti-inflammatory, anti-oxidative activities. In spite of numerous findings on its pharmacological effects, the understanding of the molecular inhibitory mechanisms of the plant remains to be studied. Therefore, we aimed to explore in vitro anti-inflammatory mechanisms of ethyl acetate fraction (Tn-EE-BF) prepared from the seed of T. nucifera in LPS-stimulated macrophage inflammatory responses. Materials and methods: For this purpose, we measured nitric oxide (NO) and prostaglandin E2 (PGE2 ) in LPS-stimulated macrophages. Additionally, using RT-PCR, luciferase reporter gene assay, immunoblotting analysis, and kinase assay, the levels of inflammatory genes, transcription factors, and inflammatory signal-regulatory proteins were investigated. Finally, the constituent of Tn-EE-BF was identified using HPLC. Results: Tn-EE-BF inhibits NO and PGE2 production and also blocks mRNA levels of inducible NO synthase (iNOS), tumor necrosis factor (TNF)-α, and cyclooxygenase (COX)-2 in a dose dependent manner. Tn-EE-BF reduces nuclear levels of the transcriptional factors NF-κB (p65) and AP-1 (c-Jun and FRA-1). Surprisingly, we found that Tn-EE-BF inhibits phosphorylation levels of Src and Syk in the NF-κB pathway, as well as, IRAK1 at the protein level, part of the AP-1 pathway. By kinase assay, we confirmed that Src, Syk, and IRAK1 are suppressed directly. HPLC analysis indicates that arctigenin, amentoflavone, and quercetin may be active components with anti-inflammatory activities. Conclusion: Tn-EE-BF exhibits anti-inflammatory activities by direct inhibition of Src/Syk/NF-κB and IRAK1/AP-1. Graphical abstract: