title: Endothelin receptor antagonists: effects on extracellular matrix synthesis in primary cultures of skin fibroblasts from systemic sclerosis patients.
abstract: Endothelin-1 (ET-1) seems to enhance the pro-fibrotic protein synthesis by skin fibroblasts and its effects are mediated by endothelin-A and B (ETA and ETB) receptors. This study aimed to investigate the effects of ETA and ETB receptor antagonists (ETARA-sitaxsentan and ETA/BRA-bosentan) on type I collagen (COL-1), fibronectin (FN) and fibrillin-1 (FBL-1) synthesis in primary cultures of skin fibroblasts from systemic sclerosis patients. Primary cultures of fibroblasts were obtained from skin biopsies of 6 female systemic sclerosis patients and were treated with ET-1 (100 nM) for 24 and 48 hrs with or without pre-treatment (1 hr) with ETARA (2 μM) or ETA/BRA (10 μM). Primary culture of human scleroderma skin fibroblasts not treated with ET-1 or ET receptor antagonists (ETARA and ETA/BRA) were used as controls. COL-1, FN and FBL-1 synthesis was evaluated by immunocytochemistry and Western blot analysis. Immunocytochemistry and Western blot analysis showed that ET-1 significantly increased COL-1 and FN synthesis at 24 and 48 hrs and FBL-1 synthesis at 48 hrs vs untreated cells. ETARA significantly contrasted the ET-1-mediated increase in COL-1 and FN at 24 hrs as well as COL-1 and FBL-1 at 48 hrs, but not FN synthesis vs ET-1-treated fibroblasts. Conversely, ETA/BRA significantly antagonized the ET-1-mediated overproduction of COL-1 and FN both at 24 and 48 hrs and the FBL-1 synthesis at 48 hrs vs ET-1-treated cells. The single ETARA treatment seems to contrast significantly the increase in COL-1 synthesis, whereas the dual ETA/BRA treatment seems active in significantly antagonizing both COL-1 and FN overproduction induced by ET-1. In conclusion, ET-1 antagonism might have positive effects in contrasting the profibrotic activity of systemic sclerosis skin fibroblasts.
authors.affiliation: Research Laboratory and Academic Unit of Clinical Rheumatology, Department of Internal Medicine, University of Genova, Genova. stefano.soldano@unige.it
authors.forename: S
authors.initials: S
authors.lastname: Soldano
authors.forename: P
authors.initials: P
authors.lastname: Montagna
authors.forename: R
authors.initials: R
authors.lastname: Brizzolara
authors.forename: C
authors.initials: C
authors.lastname: Ferrone
authors.forename: A
authors.initials: A
authors.lastname: Parodio
authors.forename: A
authors.initials: A
authors.lastname: Sulli
authors.forename: B
authors.initials: B
authors.lastname: Seriolo
authors.forename: B
authors.initials: B
authors.lastname: Villaggio
authors.forename: M
authors.initials: M
authors.lastname: Cutolo
mesh_terms: D000368:Aged; D000077300:Bosentan; D002478:Cells, Cultured; D024042:Collagen Type I / Q000096:biosynthesis; D065130:Endothelin A Receptor Antagonists; D065131:Endothelin B Receptor Antagonists; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis*; D005260:Female; D000071838:Fibrillin-1; D000071837:Fibrillins; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D005353:Fibronectins / Q000096:biosynthesis; D005355:Fibrosis; D006801:Humans; D007555:Isoxazoles / Q000494:pharmacology*; D008840:Microfilament Proteins / Q000096:biosynthesis; D008875:Middle Aged; D061251:Primary Cell Culture; D012595:Scleroderma, Systemic / Q000378:metabolism / Q000473:pathology*; D013449:Sulfonamides / Q000494:pharmacology*; D013876:Thiophenes / Q000494:pharmacology*; D054392:Adipokines
chemical_list: D000077300:Bosentan; D024042:Collagen Type I; D065130:Endothelin A Receptor Antagonists; D065131:Endothelin B Receptor Antagonists; D016326:Extracellular Matrix Proteins; D000071838:Fibrillin-1; D000071837:Fibrillins; D005353:Fibronectins; D007555:Isoxazoles; D008840:Microfilament Proteins; D013449:Sulfonamides; D013876:Thiophenes; C000605722:FBN1 protein, human; D054392:Adipokines; C106276:sitaxsentan