title: (+)-Altholactone exhibits broad spectrum immune modulating activity by inhibiting the activation of pro-inflammatory cytokines in RAW 264.7 cell lines.
abstract: An evaluation of Indonesian plants to identify compounds with immune modulating activity revealed that the methanolic extract of an Alphonsea javanica Scheff specimen possessed selective anti-inflammatory activity in a nuclear factor-kappa B (NF-κB) luciferase and MTT assay using transfected macrophage immune (Raw264.7) cells. A high-throughput LC/MS-ELSD based library approach of the extract in combination with the NF-κB and MTT assays revealed the styryl lactone (+)-altholactone (2) was responsible for the activity. Compound 2, its acetylated derivate (+)-3-O-acetylaltholactone (3), and the major compound of this class, (+)-goniothalmin (1), were further evaluated to determine their anti-inflammatory potential in the NF-κB assay. Concentration-response studies of 1-3 indicated that only 2 possessed NF-κB based anti-inflammatory activity. Compound 2 reduced the LPS-induced NO production, phosphorylation of IκBα, and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) using Western blot analysis. Further studies using qPCR indicated 2 reduced the expression of eight pro-inflammatory cytokines/enzymes (0.8-5.0μM) which included: COX-2, iNOS, IP-10, IL-1β, MCP-1, GCS-F, IL-6 and IFN-β. These results indicated that 2 displays broad spectrum immune modulating activity by functioning as an anti-inflammatory agent against LPS-induced NF-κB signaling. Conversely the selective cytotoxicity and in vivo anti-tumor and anti-inflammatory activity previously reported for 1 do not appear to arise from a mechanism that is linked to the NF-κB immune mediated pathway.
authors.affiliation: Department of Nutritional Sciences & Toxicology, University of California, Berkeley, CA 94720, USA.
authors.forename: Tyler A
authors.initials: TA
authors.lastname: Johnson
authors.forename: Johann
authors.initials: J
authors.lastname: Sohn
authors.forename: Aidan E
authors.initials: AE
authors.lastname: Ward
authors.forename: Tanya L
authors.initials: TL
authors.lastname: Cohen
authors.forename: Nicholas D
authors.initials: ND
authors.lastname: Lorig-Roach
authors.forename: Haixia
authors.initials: H
authors.lastname: Chen
authors.forename: Ronaldo A
authors.initials: RA
authors.lastname: Pilli
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Widjaja
authors.forename: Muhammad
authors.initials: M
authors.lastname: Hanafi
authors.forename: Leonardus B S
authors.initials: LB
authors.lastname: Kardono
authors.forename: Puspa D
authors.initials: PD
authors.lastname: Lotulung
authors.forename: Kyria
authors.initials: K
authors.lastname: Boundy-Mills
authors.forename: Leonard F
authors.initials: LF
authors.lastname: Bjeldanes
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D015153:Blotting, Western; D002460:Cell Line; D016207:Cytokines / Q000037:antagonists & inhibitors / Q000235:genetics; D005663:Furans / Q000037:antagonists & inhibitors*; D006801:Humans; D056747:Immunomodulation; D007249:Inflammation / Q000188:drug therapy*; D020128:Inhibitory Concentration 50; D051379:Mice; D008958:Models, Molecular; D015394:Molecular Structure; D016133:Polymerase Chain Reaction; D011753:Pyrones / Q000037:antagonists & inhibitors*; D012333:RNA, Messenger / Q000235:genetics
chemical_list: D000893:Anti-Inflammatory Agents; D016207:Cytokines; D005663:Furans; D011753:Pyrones; D012333:RNA, Messenger; C459359:altholactone