title: Daucane esters from laserwort (Laserpitium latifolium L.) inhibit cytokine and chemokine production in human lung epithelial cells.
abstract: BACKGROUND
  Laserwort, Laserpitium latifolium L. (Apiaceae), is a European medicinal plant. Its roots and rhizomes were traditionally used as a general tonic and to treat inflammatory and infective diseases.
  
  
  OBJECTIVE
  The anti-inflammatory potential of daucane esters, isolated from underground parts extract of L. latifolium and specific structural features that contribute to their activity were investigated. In addition, we studied their interference with the transactivation capacity of the Glucocorticoid Receptor when added together with a classic glucocorticoid (GC), dexamethasone (DEX). This particular property may be relevant in combination strategies, attempting to circumvent diabetogenic side effects of glucocorticoids upon long-term anti-inflammatory treatments.
  
  
  METHODS
  Nine L. latifolium daucane esters were isolated and elucidated as derivatives of desoxodehydrolaserpitin, laserpitin and a novel 2β-esterified laserpitinol analogue. Of all compounds effects on NF-κB- and AP-1-driven pro-inflammatory pathways were assessed using TNF- or PMA-induced reporter gene analysis in A549 cells. Daucanes with a strong and concentration-dependent inhibition of both NF-κB and AP-1, were tested for a potential effect on DEX-stimulated GR-driven Glucocorticoid Response Element (GRE) reporter gene activity. In addition, GRE-driven anti-inflammatory mRNA expression was determined (GILZ and DUSP1). Also anti-inflammatory properties were validated by monitoring effects on CCL-2, IL-6, IL-1β mRNA expression levels (qPCR) and on CCL-2 chemokine production (ELISA).
  
  
  RESULTS
  Daucanes featuring an ester moiety and/or a hydroxy group at positions 2β, 6α and 10α and especially the novel 2β-esterified laserpitinol derivative that, in comparison to other isolated compounds, features an additional 9α-hydroxy group, demonstrated suppression of both NF-κB- and AP-1-dependent pro-inflammatory pathways. Remarkably, those entities competitively and concentration-dependently repressed GR-driven GRE-dependent reporter gene activities. The most active compounds inhibited CCL-2 protein excretion and compound 4 downregulated genes coding for IL-1β and IL-6 induced upon TNF treatment in A549. In absence of TNF, compound 4 upregulated the GRE-mediated anti-inflammatory gene GILZ, but not DUSP1.
  
  
  CONCLUSIONS
  Daucane esters are novel anti-inflammatory agents that may, in combination with GCs, potentially improve therapeutic benefit. These results contribute to the ongoing search for novel anti-inflammatory agents as safer alternatives to, or with, GCs.
authors.affiliation: Receptor Research Laboratories, Nuclear Receptor Lab, VIB-Medical Biotechnology Center, VIB, Ghent University, Albert Baertsoenkaai 3, B-9000 Ghent, Belgium; Laboratory for Organic and Bio-Organic Synthesis, Department of Organic and Macromolecular Chemistry, Ghent University, Krijgslaan 281 S4, B-9000 Ghent, Belgium; Cancer Research Institute Ghent (CRIG), Ghent, Belgium. Electronic address: visnjap@pharmacy.bg.ac.rs.
authors.forename: Višnja
authors.initials: V
authors.lastname: Popović
authors.affiliation: Laboratory for Organic and Bio-Organic Synthesis, Department of Organic and Macromolecular Chemistry, Ghent University, Krijgslaan 281 S4, B-9000 Ghent, Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: Goeman
authors.affiliation: Receptor Research Laboratories, Nuclear Receptor Lab, VIB-Medical Biotechnology Center, VIB, Ghent University, Albert Baertsoenkaai 3, B-9000 Ghent, Belgium.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Thommis
authors.affiliation: Reliable Cancer Therapies, Boechoutlaan 221, B-1853 Strombeek-Bever, Belgium.
authors.forename: Arne
authors.initials: A
authors.lastname: Heyerick
authors.affiliation: Laboratory for Organic and Bio-Organic Synthesis, Department of Organic and Macromolecular Chemistry, Ghent University, Krijgslaan 281 S4, B-9000 Ghent, Belgium.
authors.forename: Jurgen
authors.initials: J
authors.lastname: Caroen
authors.affiliation: Laboratory for Organic and Bio-Organic Synthesis, Department of Organic and Macromolecular Chemistry, Ghent University, Krijgslaan 281 S4, B-9000 Ghent, Belgium; Cancer Research Institute Ghent (CRIG), Ghent, Belgium.
authors.forename: Johan
authors.initials: J
authors.lastname: Van der Eycken
authors.affiliation: Receptor Research Laboratories, Nuclear Receptor Lab, VIB-Medical Biotechnology Center, VIB, Ghent University, Albert Baertsoenkaai 3, B-9000 Ghent, Belgium; Department of Biochemistry, Ghent University, Albert Baertsoenkaai 3, B-9000 Ghent, Belgium; Cancer Research Institute Ghent (CRIG), Ghent, Belgium.
authors.forename: Karolien
authors.initials: K
authors.lastname: De Bosscher
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D019661:Apiaceae / Q000737:chemistry*; D018925:Chemokines / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism*; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D004952:Esters / Q000494:pharmacology*; D006801:Humans; D008168:Lung / Q000378:metabolism; D010936:Plant Extracts / Q000494:pharmacology*; D018517:Plant Roots / Q000737:chemistry; D027343:Rhizome / Q000737:chemistry; D055771:Serbia
chemical_list: D000893:Anti-Inflammatory Agents; D018925:Chemokines; D016207:Cytokines; D004952:Esters; D010936:Plant Extracts
keywords: AP-1; Anti-inflammatory activity; Chemokine production; Glucocorticoid receptor; Inducible gene expression; NF-κB