title: 8,9-Dehydrohispanolone-15,16-lactol diterpene prevents LPS-triggered inflammatory responses by inhibiting endothelial activation.
abstract: Endothelial activation contributes to lung inflammatory disorders by inducing leucocyte recruitment to pulmonary parenchyma. Consequently, vascular-targeted therapies constitute promising strategies for the treatment of inflammatory pathologies. In the present study, we evaluated the effect of 8,9-dehydrohispanolone-15,16-lactol diterpene (DT) on lung endothelium during inflammation. Lung endothelial cells pre-treated with DT and activated with lipopolysaccharide (LPS) or tumour necrosis factor-α (TNF-α) exhibited reduced expression of the pro-inflammatory cytokines Cxcl10, Ccl5 and Cxcl1, whereas the anti-inflammatory molecules IL1r2 and IL-10 were induced. Consistent with this result, DT pre-treatment inhibited nuclear factor κB (NF-κB) nuclear translocation, by interfering with IκBα phosphorylation, and consequently NF-κB transcriptional activity in endothelium activated by LPS or TNF-α. Furthermore, DT, probably through p38 signalling, induced transcriptional activation of genes containing activator protein 1 (AP-1)-binding elements. Inhibition of p38 prevented IL1r2 mRNA expression in endothelium incubated with DT alone or in combination with LPS or TNF-α. Accordingly, conditioned medium (CM) from these cells failed to stimulate leucocytes as measured by a reduction in adhesive ability of the leucocyte cell line J774 to fibronectin (FN). Additionally, DT reduced the expression of the endothelial adhesion molecules E-selectin, vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) after activation. Similarly, expression of VCAM-1 and ICAM-1 molecules on the lung endothelial layer of C57/BL6 mice pre-treated with DT and challenged with LPS were unchanged. Finally, inhibition of vascular adhesion molecule expression by DT decreased the interaction of J774 cells with lung endothelial cells in an inflammatory environment. Our findings establish DT as a novel endothelial inhibitor for the treatment of inflammatory-related diseases triggered by Gram-negative bacteria or by the associated cytokine TNF-α.
authors.affiliation: Unidad de Terapias Farmacológicas, Área de Genética Humana, Instituto de Investigación de Enfermedades Raras (IIER), Instituto de Salud Carlos III, 28220, Majadahonda, Spain.
authors.forename: Lidia
authors.initials: L
authors.lastname: Jiménez-García
authors.affiliation: Molecular Neurobiology Laboratory, The Salk Institute, La Jolla, CA 92037, U.S.A.
authors.forename: Paqui G
authors.initials: PG
authors.lastname: Través
authors.affiliation: Departamento de Farmacia y Biotecnología, Facultad de Ciencias Biomédicas, Universidad Europea de Madrid, 28670, Villaviciosa de Odón, Spain.
authors.forename: Raquel
authors.initials: R
authors.lastname: López-Fontal
authors.affiliation: Unidad de Terapias Farmacológicas, Área de Genética Humana, Instituto de Investigación de Enfermedades Raras (IIER), Instituto de Salud Carlos III, 28220, Majadahonda, Spain.
authors.forename: Sandra
authors.initials: S
authors.lastname: Herranz
authors.affiliation: Unidad de Terapias Farmacológicas, Área de Genética Humana, Instituto de Investigación de Enfermedades Raras (IIER), Instituto de Salud Carlos III, 28220, Majadahonda, Spain.
authors.forename: María Angeles
authors.initials: MA
authors.lastname: Higueras
authors.affiliation: Departamento de Farmacología, Facultad de Farmacia, Universidad Complutense de Madrid, 28040, Madrid, Spain.
authors.forename: Beatriz
authors.initials: B
authors.lastname: de Las Heras
authors.affiliation: Unidad de Terapias Farmacológicas, Área de Genética Humana, Instituto de Investigación de Enfermedades Raras (IIER), Instituto de Salud Carlos III, 28220, Majadahonda, Spain shortelano@isciii.es aluque@isciii.es.
authors.forename: Sonsoles
authors.initials: S
authors.lastname: Hortelano
authors.affiliation: Unidad de Terapias Farmacológicas, Área de Genética Humana, Instituto de Investigación de Enfermedades Raras (IIER), Instituto de Salud Carlos III, 28220, Majadahonda, Spain shortelano@isciii.es aluque@isciii.es.
authors.forename: Alfonso
authors.initials: A
authors.lastname: Luque
mesh_terms: D000818:Animals; D002460:Cell Line; D018946:Chemokine CCL5 / Q000378:metabolism; D054360:Chemokine CXCL1 / Q000378:metabolism; D054357:Chemokine CXCL10 / Q000378:metabolism; D004224:Diterpenes / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism*; D007249:Inflammation / Q000139:chemically induced / Q000378:metabolism / Q000517:prevention & control*; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
chemical_list: D018946:Chemokine CCL5; D054360:Chemokine CXCL1; D054357:Chemokine CXCL10; D004224:Diterpenes; D008070:Lipopolysaccharides; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1
keywords: adhesion molecules; cytokines; diterpenoid derivatives; endothelial cells; inflammation