title: Anti-inflammatory constituents from the root of Litsea cubeba in LPS-induced RAW 264.7 macrophages.
abstract: Context Litsea cubeba (Lour.) Pers. (Lauraceae) has long been used as a folk remedy in Traditional Chinese Medicine (TCM) for the treatment of rheumatic diseases. Previous studies from our laboratory indicated that L. cubeba extract showed anti-arthritic activity in rats. Objective To study L. cubeba chemically and biologically and to find the potential constituents responsible for its anti-arthritic effect. Materials and methods The compounds were isolated from the root of L. cubeba by column chromatography which eluted with PE:EtOAc gradient system, and the structures were elucidated by detailed spectroscopic data analysis; the anti-inflammatory activity of the isolated compounds was evaluated by lipopolysaccharide (LPS)-induced RAW 264.7 cells and the TNF-α and NO level were measured by ELISA (commercial kit); The iNOS and COX-2 mRNA expression were measured by RT-PCR and the phosphorylation of IκBα, IKKβ, P38 and Akt were determined by western blots. Results A novel 9-fluorenone, 1-ethoxy-3,7-dihydroxy-4,6-dimethoxy-9-fluorenone (1), together with 4 known compounds, namely pinoresinol (2), syringaresinol (3), 9,9'-O-di-(E)-feruloyl-meso-5,5'-dimethoxysecoisolariciresinol (4) and lyoniresinol (5) were isolated from the root of L. cubeba for the first time. The IC50 for NO inhibition on compounds 1 and 4 were 56.1 ± 1.2 and 32.8 ± 2.3 μM, respectively. The IC50 for TNF-α inhibition were 28.2 ± 0.9 and 15.0 ± 1.0 μM, respectively. Both 1 and 4 suppress mRNA expression of iNOS, COX-2 and protein phosphorylation of IκBα, IKKβ in LPS-induced RAW 264.7 cells. Discussion and conclusion Compounds 1 and 4 isolated from L. cubeba exhibited potent anti-inflammatory activity through the NF-κB signal pathway.
authors.affiliation: a School of Pharmacy , Second Military Medical University , Shanghai , China ;|b Department of Pharmacy , Fuzhou General Hospital of Nanjing Military Region , Fuzhou , China ;
authors.forename: Bing
authors.initials: B
authors.lastname: Lin
authors.affiliation: a School of Pharmacy , Second Military Medical University , Shanghai , China ;
authors.forename: Lian-Na
authors.initials: LN
authors.lastname: Sun
authors.affiliation: a School of Pharmacy , Second Military Medical University , Shanghai , China ;
authors.forename: Hai-Liang
authors.initials: HL
authors.lastname: Xin
authors.affiliation: c Department of Pharmacy , Yueyang Hospital of Integrated Traditional Chinese Medicine and Western Medicine, Shanghai University of Traditional Chinese Medicine , Shanghai , China.
authors.forename: Hua
authors.initials: H
authors.lastname: Nian
authors.affiliation: b Department of Pharmacy , Fuzhou General Hospital of Nanjing Military Region , Fuzhou , China ;
authors.forename: Hong-Tao
authors.initials: HT
authors.lastname: Song
authors.affiliation: a School of Pharmacy , Second Military Medical University , Shanghai , China ;
authors.forename: Yi-Ping
authors.initials: YP
authors.lastname: Jiang
authors.affiliation: a School of Pharmacy , Second Military Medical University , Shanghai , China ;
authors.forename: Zhen-Qiao
authors.initials: ZQ
authors.lastname: Wei
authors.affiliation: a School of Pharmacy , Second Military Medical University , Shanghai , China ;
authors.forename: Lu-Ping
authors.initials: LP
authors.lastname: Qin
authors.affiliation: a School of Pharmacy , Second Military Medical University , Shanghai , China ;
authors.forename: Ting
authors.initials: T
authors.lastname: Han
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000302:isolation & purification / Q000494:pharmacology*; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D007249:Inflammation / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000517:prevention & control*; D018836:Inflammation Mediators / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D032862:Litsea / Q000737:chemistry; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D015394:Molecular Structure; D016328:NF-kappa B / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D008517:Phytotherapy; D018517:Plant Roots; D010946:Plants, Medicinal; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D000067996:RAW 264.7 Cells; D015398:Signal Transduction / Q000187:drug effects; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D016207:Cytokines; D018836:Inflammation Mediators; D008070:Lipopolysaccharides; D016328:NF-kappa B; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; C077623:Ptgs2 protein, mouse; D051546:Cyclooxygenase 2; D051057:Proto-Oncogene Proteins c-akt; D048051:p38 Mitogen-Activated Protein Kinases
keywords: Anti-arthritic; IKKβ; IκBα; RT-PCR; Traditional Chinese Medicine; rheumatoid arthritis