title: Anti-inflammatory activity of standardized dichloromethane extract of Salvia connivens on macrophages stimulated by LPS.
abstract: BACKGROUND
  A previous study demonstrated that the chloroform extract of Salvia connivens Epling (Lamiaceae) has anti-inflammatory activity.
  
  
  OBJECTIVE
  Identification of the active components in the dicholorometane extract (DESC), and, standardization of the extract based in ursolic acid.
  
  
  METHODS
  DESC was prepared by percolation with dichlromethane and after washed with hot hexane, its composition was determined by CG-MS and NMR, and standardized by HPLC. The anti-inflammatory activity was tested on acute TPA-induced mouse ear oedema at doses of 2.0 mg/ear. The cell viability of macrophages was evaluated by MTT method, and pro- and anti-inflammatory interleukin levels were measured using an ELISA kit.
  
  
  RESULTS
  Ursolic acid, oleanolic acid, dihydroursolic acid and eupatorin were identified in DESC, which was standardized based on the ursolic acid concentration (126 mg/g). The anti-inflammatory activities of DESC, the acid mixture, and eupatorin (2 mg/ear) were 60.55, 57.20 and 56.40% inhibition, respectively, on TPA-induced ear oedema. The IC50of DESC on macrophages was 149.4 μg/mL. DESC (25 μg/mL) significantly reduced TNF-α (2.0-fold), IL-1β (2.2-fold) and IL-6 (2.0-fold) in macrophages stimulated with LPS and increased the production of IL-10 (1.9-fold).
  
  
  CONCLUSIONS
  Inflammation is a basic response to injuries, and macrophages are involved in triggering inflammation. Macrophage cells exhibit a response to LPS, inducing inflammatory mediators, and DESC inhibits the biosynthesis of the pro-inflammatory and promote anti-inflammatory cytokines.
  
  
  CONCLUSIONS
  DESC has an anti-inflammatory effect; reduced the levels of IL-1β, Il-6 and TNF-α; and increases IL-10 in macrophages stimulated with LPS. Ursolic acid is a good phytochemical marker.
authors.affiliation: a Facultad de Ciencias Químicas , Centro de Investigación y Estudios de Posgrado, Universidad Autónoma de San Luis Potosí , San Luis Potosí , SLP , México.
authors.forename: Marco Martín
authors.initials: MM
authors.lastname: González-Chávez
authors.affiliation: a Facultad de Ciencias Químicas , Centro de Investigación y Estudios de Posgrado, Universidad Autónoma de San Luis Potosí , San Luis Potosí , SLP , México.
authors.forename: Cinthia Saraí
authors.initials: CS
authors.lastname: Ramos-Velázquez
authors.affiliation: b Departamento de Sistemas Biológicos , Universidad Autónoma Metropolitana - Xochimilco , Ciudad de México , México.
authors.forename: Roberto
authors.initials: R
authors.lastname: Serrano-Vega
authors.affiliation: b Departamento de Sistemas Biológicos , Universidad Autónoma Metropolitana - Xochimilco , Ciudad de México , México.
authors.forename: Cuauhtemoc
authors.initials: C
authors.lastname: Pérez-González
authors.affiliation: b Departamento de Sistemas Biológicos , Universidad Autónoma Metropolitana - Xochimilco , Ciudad de México , México.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Sánchez-Mendoza
authors.affiliation: b Departamento de Sistemas Biológicos , Universidad Autónoma Metropolitana - Xochimilco , Ciudad de México , México.
authors.forename: Salud
authors.initials: S
authors.lastname: Pérez-Gutiérrez
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000302:isolation & purification / Q000494:pharmacology*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004487:Edema / Q000139:chemically induced / Q000276:immunology / Q000517:prevention & control*; D005419:Flavonoids / Q000302:isolation & purification / Q000494:pharmacology; D008401:Gas Chromatography-Mass Spectrometry; D053583:Interleukin-1beta / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008262:Macrophage Activation / Q000187:drug effects*; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008752:Methylene Chloride / Q000737:chemistry*; D051379:Mice; D009828:Oleanolic Acid / Q000302:isolation & purification / Q000494:pharmacology; D008517:Phytotherapy; D035261:Plant Components, Aerial / Q000737:chemistry; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D010946:Plants, Medicinal; D027544:Salvia / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry*; D013755:Tetradecanoylphorbol Acetate; D014315:Triterpenes / Q000302:isolation & purification / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D000097245:Ursolic Acid
chemical_list: D000893:Anti-Inflammatory Agents; D005419:Flavonoids; D053583:Interleukin-1beta; D015850:Interleukin-6; D008070:Lipopolysaccharides; D008752:Methylene Chloride; D009828:Oleanolic Acid; D010936:Plant Extracts; D012997:Solvents; D013755:Tetradecanoylphorbol Acetate; D014315:Triterpenes; D014409:Tumor Necrosis Factor-alpha; D000097245:Ursolic Acid; C587672:IL1B protein, mouse; C577686:interleukin-6, mouse; C103110:eupatorin
keywords: HPLC; Ursolic acid; cellular viability; cytokine determination; eupatorin