title: Ethanolic extract and water-soluble polysaccharide from Chaenomeles speciosa fruit modulate lipopolysaccharide-induced nitric oxide production in RAW264.7 macrophage cells.
abstract: BACKGROUND
  Chaenomeles speciosa fruits have been widely used in traditional Chinese medicine for treatment of diseases related to inflammatory reaction. This study aims to identify anti-inflammatory and immunomodulatory components of Chaenomeles speciosa fruit and unravel their potential mechanisms.
  
  
  METHODS
  Ethanolic extract and its n-hexane, chloroform, ethyl acetate and n-butanol fractions, as well as water-soluble polysaccharide, were prepared from dry fruits of Chaenomeles speciosa. The mouse macrophage-like RAW264.7 cells were induced by lipopolysaccharide (LPS) and used as an inflammatory cell model. Production of nitric oxide in the cells was determined by the Griess assay, and cell viability was tested by the MTT method. Cellular apoptosis was evaluated by fluorescence-activated cell sorting. Relative quantification of inflammation-related genes was analyzed by real-time PCR.
  
  
  RESULTS
  LPS-induced production of nitric oxide in RAW264.7 cells was significantly inhibited by the ethyl acetate fraction (EAF) at 200-800μg/ml, while Chaenomeles speciosa polysaccharide (CPS) promoted nitric oxide production at 250-750μg/ml either alone or in an additive fashion with LPS. Both EAF and CPS did not provoke noticeable cytotoxicity and apoptosis at the above effective concentrations. EAF significantly reduced LPS-induced upshift of iNOS mRNA level but showed no significant effect on the induction of IFN-γ and G-CSF, while CPS reduced the gene induction of TNF-α, IFN-γ and G-CSF by LPS.
  
  
  CONCLUSIONS
  EAF was able to inhibit nitric oxide production by reducing LPS-induced upshift of iNOS mRNA level. CPS was an activator of nitric oxide production through cytokines such as TNF-α, IFN-γ and G-CSF. These results demonstrate the therapeutic effects of both ethanolic and aqueous extracts of Chaenomeles speciosa fruit, a traditional edible medicine used in health maintenance and disease treatment.
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, and School of Pharmaceutical Sciences, Wuhan University, Wuhan, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhu
authors.forename: Chaolin
authors.initials: C
authors.lastname: Liao
authors.forename: Yongmei
authors.initials: Y
authors.lastname: Liu
authors.forename: Pengcheng
authors.initials: P
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Guo
authors.forename: Meijun
authors.initials: M
authors.lastname: He
authors.forename: Zebo
authors.initials: Z
authors.lastname: Huang
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002460:Cell Line; D016207:Cytokines / Q000235:genetics; D000431:Ethanol / Q000737:chemistry; D005638:Fruit; D008070:Lipopolysaccharides; D008264:Macrophages / Q000187:drug effects / Q000276:immunology; D008516:Medicine, Chinese Traditional; D051379:Mice; D009569:Nitric Oxide / Q000276:immunology; D010936:Plant Extracts / Q000494:pharmacology*; D011134:Polysaccharides / Q000737:chemistry / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D027824:Rosaceae; D012995:Solubility; D012997:Solvents / Q000737:chemistry; D014867:Water / Q000737:chemistry
chemical_list: D000893:Anti-Inflammatory Agents; D016207:Cytokines; D008070:Lipopolysaccharides; D010936:Plant Extracts; D011134:Polysaccharides; D012333:RNA, Messenger; D012997:Solvents; D014867:Water; D009569:Nitric Oxide; D000431:Ethanol