title: Bioassay-guided fractionation of ethyl acetate extract from Armillaria mellea attenuates inflammatory response in lipopolysaccharide (LPS) stimulated BV-2 microglia.
abstract: BACKGROUND
  Armillaria mellea (A. mellea) is a traditional Chinese medicinal and edible mushroom, which is proved to possess a lot of biological activities, including anti-oxidation, immunopotentiation, anti-vertigo and anti-aging activities. However, little information is available in regard to its neuroprotection activity in inflammation-mediated neurodegenerative diseases.
  
  
  OBJECTIVE
  We have found that A. mellea has an anti-inflammatory activity in LPS-induced RAW264.7 cells in our previous study. The objective of this study is to investigate the anti-neuroinflammatory mechanism of a bioassay-guided fractionation (Fr.2) and its active components/compounds.
  
  
  METHODS
  Compounds were isolated by preparative high performance liquid chromatography (pre-HPLC) and their structures were established by mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopic analyses. The anti-neuroinflammatory effect of Fr.2 and each compounds were investigated in lipopolysaccharide (LPS)-stimulated murine microglia cell lineBV-2.
  
  
  RESULTS
  We demonstrated that Fr.2 significantly decreased the production of inflammation mediator nitric oxide (NO) and inflammatory cytokines tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1beta (IL-1β) in a dose-dependent manner (10, 30, 100µg/ml). In addition, Fr.2 markedly down-regulated the phosphorylation levels of nuclear factor kappa B p65 (NF-κB p65), inhibitory κB-α (IκB-α) and c-Jun N-terminal kinases (JNKs) pathways. Sevens compounds were isolated from Fr.2, among them, three compounds, 5-hydroxymethylfurfural (CP1), vanillic acid (CP4) and syringate (CP5) were reported for the first time in A. mellea. NO and inflammatory cytokines (TNF-α, IL-6, IL-1β) secretion indicated that daidzein (CP6) and genistein (CP7) showed a more outstanding anti-inflammation potential at non-toxic concentrations (10, 30, 100µM) than the other five compounds.
  
  
  CONCLUSIONS
  In conclusion, Fr.2 may have therapeutic potential for neurodegenerative diseases by inhibiting inflammatory mediators and suppress inflammation pathway in activated microglia. Daidzein and genistein may serve as the effective anti-inflammation compounds of Fr.2.
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Geng
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China.
authors.forename: Shuiling
authors.initials: S
authors.lastname: Zhu
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Cheng
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China.
authors.forename: Zhen-Ming
authors.initials: ZM
authors.lastname: Lu
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China.
authors.forename: Hong-Yu
authors.initials: HY
authors.lastname: Xu
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China.
authors.forename: Jin-Song
authors.initials: JS
authors.lastname: Shi
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China. Electronic address: zhenghxu@jiangnan.edu.cn.
authors.forename: Zheng-Hong
authors.initials: ZH
authors.lastname: Xu
mesh_terms: D000085:Acetates / Q000494:pharmacology*; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D055392:Armillaria / Q000737:chemistry; D004365:Drugs, Chinese Herbal / Q000494:pharmacology; D007249:Inflammation / Q000188:drug therapy*; D008070:Lipopolysaccharides / Q000009:adverse effects*; D051379:Mice; D017628:Microglia / Q000187:drug effects*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D010936:Plant Extracts / Q000494:pharmacology*
chemical_list: D000085:Acetates; D000893:Anti-Inflammatory Agents; D004365:Drugs, Chinese Herbal; D008070:Lipopolysaccharides; D018696:Neuroprotective Agents; D010936:Plant Extracts; C007650:ethyl acetate
keywords: Anti-inflammation; Armillaria mellea; BV-2 microglia; Neurodegenerative diseases; Neuroinflammation