title: Further evidence for the anti-inflammatory activity of oleocanthal: inhibition of MIP-1α and IL-6 in J774 macrophages and in ATDC5 chondrocytes.
abstract: OBJECTIVE
  Given the relevance of degenerative joint diseases in our society, the development of a novel pharmacologic intervention is a critically important public health goal. Recently, oleocanthal, a polyphenolic natural compound from extra virgin olive oil, has emerged as a potential therapeutic weapon for the treatment of inflammatory degenerative diseases. The goal of this study was to further evaluate the anti-inflammatory activity of oleocanthal in murine macrophages J774 and murine chondrocytes ATDC5 with a particular focus on the inhibition of gene expression of pro-inflammatory factors such as MIP-1α and IL-6.
  
  
  METHODS
  ATDC5 murine chondrogenic cells and murine macrophages J774 were used. J774 macrophages were tested with different doses of oleocanthal and cell viability was evaluated using the MTT assay. Western blot analysis was carried on in J774 cells using anti NOS2 antibody. Nitrite accumulation was determined in culture supernatant using the Griess reaction. MIP-1α and IL-6 mRNA levels were determined using SYBR Green-based quantitative RT-PCR. MIP-1α and IL-6 protein levels were evaluated using specific ELISA assay. Several cytokines, involved in the inflammatory response, were also tested by BioPlex assay.
  
  
  RESULTS
  First, oleocanthal inhibits LPS-induced NO production in J774 macrophages, without affecting cell viability. Moreover, it inhibits MIP-1α and IL-6 mRNA expression, as well as protein synthesis, in both ATDC5 chondrocytes and J774 macrophages. Oleocanthal also inhibits IL-1β, TNF-α and GM-CSF protein synthesis from LPS-stimulated macrophages.
  
  
  CONCLUSIONS
  Our data confirm a clear potent role of oleocanthal as anti-inflammatory therapeutic agent for future treatment of arthritis or other inflammatory diseases.
authors.affiliation: SERGAS, Santiago University Clinical Hospital Research Laboratory 9 (NEIRID LAB: Neuroendocrine Interactions in Rheumatology and Inflammatory Diseases), Institute of Medical Research (IDIS), Santiago de Compostela 15706, Spain.
authors.forename: Morena
authors.initials: M
authors.lastname: Scotece
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Gómez
authors.forename: Javier
authors.initials: J
authors.lastname: Conde
authors.forename: Verónica
authors.initials: V
authors.lastname: Lopez
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Gómez-Reino
authors.forename: Francisca
authors.initials: F
authors.lastname: Lago
authors.forename: Amos B
authors.initials: AB
authors.lastname: Smith
authors.forename: Oreste
authors.initials: O
authors.lastname: Gualillo
mesh_terms: D000447:Aldehydes / Q000737:chemistry / Q000494:pharmacology*; D000704:Analysis of Variance; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D052160:Benzothiazoles; D015153:Blotting, Western; D054405:Chemokine CCL3 / Q000037:antagonists & inhibitors*; D019902:Chondrocytes / Q000378:metabolism*; D000081004:Cyclopentane Monoterpenes; D003959:Diamines; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation / Q000187:drug effects*; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D015394:Molecular Structure; D009930:Organic Chemicals; D010636:Phenols / Q000737:chemistry / Q000494:pharmacology*; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction; D013778:Tetrazolium Salts; D013844:Thiazoles; D000093242:Interleukin-6 Inhibitors
chemical_list: D000447:Aldehydes; D000893:Anti-Inflammatory Agents; D052160:Benzothiazoles; D054405:Chemokine CCL3; D000081004:Cyclopentane Monoterpenes; D003959:Diamines; D009930:Organic Chemicals; D010636:Phenols; D011804:Quinolines; D013778:Tetrazolium Salts; D013844:Thiazoles; D000093242:Interleukin-6 Inhibitors; C098022:SYBR Green I; C503534:oleocanthal; C022616:thiazolyl blue