title: Evaluation of anti-inflammatory activity of compounds isolated from the rhizome of Ophiopogon japonicas.
abstract: BACKGROUND
  Ophiopogon japonicas (L.f) Ker-Gawl has been used as a traditional Chinese medicine to cure acute and chronic inflammation and cardiovascular diseases including thrombotic diseases for thousands of years. Previous phytochemical studies showed that O. japonicus contained compounds with anti-inflammatory activity. The aim of this study was to identify and isolate compounds with anti-inflammatory activity from the rhizome of O. japonicas.
  
  
  METHODS
  Compounds were isolated by various column chromatography and their structures were identified in terms of nuclear magnetic resonance spectrum (NMR) and mass spectrum (MS). To measure the anti-inflammatory effects of thirteen compounds in LPS-induced RAW 264.7 macrophage cells, we used the following methods: cell viability assay, nitric oxide assay, enzyme-linked immunosorbent assay, quantitative real-time PCR analysis and western blotting analysis.
  
  
  RESULTS
  One new and twelve known compounds (mainly homoisoflavonoids) were extracted from O. japonicas, in which 4'-O-Demethylophiopogonanone E (10) was considered as a new compound, additionally, compounds 4-O-(2-Hydroxy-1- hydroxymethylethyl)-dihydroconiferyl alcohol (2) and 5,7-dihydroxy-6-methyl-3-(2', 4'-dihydroxybenzyl) chroman-4-one (12) were isolated from the rhizome of O. japonicas for the first time. The isolated compounds Oleic acid (3), Palmitic acid (4), desmethylisoophiopogonone B [5,7-dihydroxy-3-(4'-hydroxybenzyl)-8- methyl- chromone] (5), 5,7-dihydroxy-6-methyl-3-(4'-hydroxybenzyl) chromone (7) and 10 significantly suppressed the production of NO in LPS-induced RAW 264.7 cells. Especially compound 10 showed the strongest effect against the production of the pro-inflammatory cytokine IL-1β and IL-6 with the IC50value of 32.5 ± 3.5 μg/mL and 13.4 ± 2.3 μg/mL, respectively. Further analysis elucidated that the anti-inflammatory activity of compound 10 might be exerted through inhibiting the phosphorylation of ERK1/2 and JNK in MAPK signaling pathways to decrease NO and pro-inflammatory cytokines production.
  
  
  CONCLUSIONS
  Our results indicated that 4'-O-Demethylophiopogonanone E can be considered as a potential source of therapeutic medicine for inflammatory diseases.
authors.affiliation: The Second Institute of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou, 510405, Guangdong, China.
authors.forename: Jing-Wen
authors.initials: JW
authors.lastname: Zhao
authors.affiliation: Guangdong Food and Drug Vocational College, Guangzhou, 510520, Guangdong, China.
authors.forename: Ding-Sheng
authors.initials: DS
authors.lastname: Chen
authors.affiliation: Artepharm Company Limited, Guangzhou, 510410, Guangdong, China.
authors.forename: Chang-Sheng
authors.initials: CS
authors.lastname: Deng
authors.affiliation: Artepharm Company Limited, Guangzhou, 510410, Guangdong, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: The Second Institute of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou, 510405, Guangdong, China. zhuwei9201@163.com.
authors.forename: Wei
authors.identifier: http://orcid.org/0000-0002-8200-9230
authors.initials: W
authors.lastname: Zhu
authors.affiliation: Artepharm Company Limited, Guangzhou, 510410, Guangdong, China. linli0921@21cn.com.
authors.forename: Li
authors.initials: L
authors.lastname: Lin
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000737:chemistry* / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000494:pharmacology*; D006801:Humans; D007249:Inflammation / Q000188:drug therapy / Q000235:genetics / Q000276:immunology* / Q000503:physiopathology; D053583:Interleukin-1beta / Q000235:genetics / Q000276:immunology; D008264:Macrophages / Q000187:drug effects / Q000276:immunology; D051379:Mice; D015394:Molecular Structure; D016328:NF-kappa B / Q000235:genetics / Q000276:immunology; D031441:Ophiopogon / Q000737:chemistry*; D000067996:RAW 264.7 Cells; D027343:Rhizome / Q000737:chemistry
chemical_list: D000893:Anti-Inflammatory Agents; D004365:Drugs, Chinese Herbal; D053583:Interleukin-1beta; D016328:NF-kappa B
keywords: Anti-inflammatory; Homoisoflavonoids; MAPKs; Macrophages; Ophiopogon japonicus