{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2022,3,29]],"date-time":"2022-03-29T02:33:06Z","timestamp":1648521186559},"reference-count":33,"publisher":"The Journal of Rheumatology","issue":"3","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J Rheumatol"],"published-print":{"date-parts":[[2015,3]]},"abstract":"<jats:sec><jats:title>Objective.<\/jats:title><jats:p>To investigate the effects of the endothelin 1 (ET-1) receptor antagonists (ETRA) macitentan, its active metabolite ACT-132577, and bosentan on myofibroblast activation and extracellular matrix production induced by ET-1 in cultured systemic sclerosis (SSc) and control skin fibroblasts.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods.<\/jats:title><jats:p>Fibroblasts were obtained from skin biopsies of 6 patients with SSc and 5 healthy subjects. Some cultured cells were untreated or treated with macitentan, ACT-132577, or bosentan alone (10 \u03bcM). Other cultured cells were treated with ET-1 alone (100 nM) or with ETRA, and after 1 h, also with ET-1. After 48 h of treatment, myofibroblast activation was investigated to evaluate the \u03b1-smooth muscle actin (\u03b1-SMA) expression by immunofluorescence; type I collagen (COL-1) and fibronectin (FN) were investigated by immunocytochemistry, Western blotting, and quantitative real-time PCR (qRT-PCR). Statistical analysis was performed by the nonparametric Mann-Whitney U test.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results.<\/jats:title><jats:p>In cultured SSc skin fibroblasts, only the treatment with macitentan significantly reduced the basal level of \u03b1-SMA expression (p = 0.03 vs untreated cells). Macitentan also significantly reduced the basal level of COL-1 synthesis, similarly to bosentan (p &lt; 0.05 vs untreated cells). Macitentan or ACT-132577 antagonized the ability of ET-1 to further induce \u03b1-SMA expression (p = 0.03), COL-1, and FN synthesis (p = 0.03, p = 0.005); bosentan showed similar effects. These results obtained by immunofluorescence and immunocytochemistry were confirmed by Western blotting and qRT-PCR. The downregulatory effects exerted by ETRA were observed also in cultured human control skin fibroblasts.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion.<\/jats:title><jats:p>Macitentan and ACT-132577 seem to downregulate <jats:italic>in vitro<\/jats:italic> the profibrotic myofibroblast phenotype induced by ET-1 in cultured human SSc skin fibroblasts.<\/jats:p><\/jats:sec>","DOI":"10.3899\/jrheum.141070","type":"journal-article","created":{"date-parts":[[2015,1,16]],"date-time":"2015-01-16T03:16:23Z","timestamp":1421378183000},"page":"456-463","source":"Crossref","is-referenced-by-count":10,"title":["Effects of Macitentan and Its Active Metabolite on Cultured Human Systemic Sclerosis and Control Skin 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