title: Bioactivity-guided isolation of polyacetylenes with inhibitory activity against NO production in LPS-activated RAW264.7 macrophages from the rhizomes of Atractylodes macrocephala.
abstract: BACKGROUND
  The rhizome of Atractylodes macrocephala (Compositae) is one of the most well-known traditional Chinese medicine in China, Japan and Korea, which has a long history of use for the treatment of splenic asthenia, edema, anorexia, and excessive perspiration, etc. As active compounds of anti-inflammatory activity of this medicinal plant have not been fully elucidated, the aim of this study was to isolate and identify the active constituents inhibiting nitric oxide (NO) production from the rhizomes of A. macrocephala.
  
  
  METHODS
  Inhibitory activity against NO production in lipopolysaccharide-activated RAW264.7 macrophages was evaluated by Griess reaction. Fifteen polyacetylenes were isolated from the active ethyl acetate extract using activity-guided screening. The structures of all compounds were elucidated by spectroscopic methods and comparison with published data. The compounds were further tested for their inhibitory activity against NO production.
  
  
  RESULTS
  Seven new polyacetylenes, named atractylodemaynes A-G (1-7), along with eight known ones (8-15) were isolated. Compound 14 was isolated for the first time from the rhizomes of A. macrocephala. The study showed that the tested compounds exhibited inhibitory activity against NO production in a dose-dependent manner. Among them, compounds 10, 11 and 12 had relatively stronger inhibitory effect with IC50 values of 28, 23 and 19μM, respectively.
  
  
  CONCLUSIONS
  The results demonstrated that the polyacetylenes might greatly contribute to the anti-inflammatory activity of the rhizomes of A. macrocephala.
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs (Peking University), Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University Health Science Center, Peking University, Beijing 100191, China.
authors.forename: Chun-Mei
authors.initials: CM
authors.lastname: Yao
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs (Peking University), Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University Health Science Center, Peking University, Beijing 100191, China. Electronic address: xwyang@bjmu.edu.cn.
authors.forename: Xiu-Wei
authors.initials: XW
authors.lastname: Yang
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D031174:Atractylodes; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D008070:Lipopolysaccharides; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D009569:Nitric Oxide / Q000037:antagonists & inhibitors*; D010936:Plant Extracts / Q000737:chemistry; D053279:Polyynes / Q000494:pharmacology*; D027343:Rhizome / Q000737:chemistry*
chemical_list: D000893:Anti-Inflammatory Agents; D008070:Lipopolysaccharides; D010936:Plant Extracts; D053279:Polyynes; D009569:Nitric Oxide
keywords: (1)H–(1)H COSY; (1)H–(1)H correlated spectroscopy; 14-Acetoxy-12-senecioyloxytetradeca-2E,8E,10E-trien-4,6-diyn-1-ol (PubChem CID: 14448076); 14-Acetoxy-12-α-methylbutyryltetradeca-2E,8E,10E-trien-4,6-diyne-1-ol (PubChem CID: 5319529); 14-Acetoxy-12-α-methylbutyryltetradeca-2E,8Z,10E-trien-4,6-diyne-1-ol (PubChem CID: 5319530); 14-Acetoxy-12-β-methylbutyryltetradeca-2E,8E,10E-trien-4,6-diyne-1-ol (PubChem CID: 14586258); 14-α-Methylbutyryltetradeca-2E,8E,10E-trien-4,6-diyne-1-ol (PubChem CID: 5319531); 14-β-Methylbutyryltetradeca-2E,8E,10E-trien-4,6-diyne-1-ol (PubChem CID: 11544212); 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide; AMR; AO-I; AO-III; Anti-inflammatory; Atractylodes macrocephala; Atractylodis Macrocephalae Rhizoma; BuOH; CC; CHX; Compositae; EtOAc; EtOH; HMBC; HPLC; HRESIMS; HSQC; IND; L-N(6)-(1-iminoethyl)-lysine; L-NIL; LPS; MTT; NMR; NO; Nitric oxide; Polyacetylenes; atractylenolide I; atractylenolide III; column chromatography; cyclohexane; ethanol; ethyl acetate; heteronuclear multiple bond correlation; heteronuclear single quantum correlation; high-performance liquid chromatography; high-resolution electric spray ion mass spectrum; indomethacin; lipopolysaccharides; n-butanol; nitric oxide; nuclear magnetic resonance