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We previously reported dexamethasone enhanced the pro-fibrotic effects of transforming growth factor (TGF)-\u03b2 as a potential mechanism for inconsistent clinical outcomes. In the current study, we sought to determine the mechanism(s) whereby GCs influence the fibrotic response and mechanisms underlying these effects with an emphasis on TGF-\u03b2 and nuclear receptor subfamily 4 group A member 1 (NR4A1) signaling. Human VF fibroblasts (HVOX) were treated with three commonly-employed GCs+\u2009\/-TGF-\u03b21. Phosphorylation of the glucocorticoid receptor (GR:NR3C1) and activation of NR4A1 was analyzed by western blotting. Genes involved in the fibrotic response, including <jats:italic>ACTA2, TGFBR1,<\/jats:italic> and <jats:italic>TGFBR2<\/jats:italic> were analyzed by qPCR. RNA-seq was performed to identify global changes in gene expression induced by dexamethasone. GCs enhanced phosphorylation of GR at Ser211 and TGF-\u03b2-induced <jats:italic>ACTA2<\/jats:italic> expression. Dexamethasone upregulated <jats:italic>TGFBR1<\/jats:italic>, and <jats:italic>TGFBR2<\/jats:italic> in the presence of TGF-\u03b21 and increased active NR4A1. RNA-seq results confirmed numerous pathways, including TGF-\u03b2 signaling, affected by dexamethasone. Synergistic pro-fibrotic effects of TGF-\u03b2 were observed across GCs and appeared to be mediated, at least partially, via upregulation of TGF-\u03b2 receptors. Dexamethasone exhibited diverse regulation of gene expression including NR4A1 upregulation consistent with the anti-fibrotic potential of GCs.<\/jats:p>","DOI":"10.1038\/s41598-020-77445-9","type":"journal-article","created":{"date-parts":[[2020,11,24]],"date-time":"2020-11-24T11:16:29Z","timestamp":1606216589000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Complex fibroblast response to glucocorticoids may underlie variability of clinical efficacy in the vocal folds"],"prefix":"10.1038","volume":"10","author":[{"given":"Ryosuke","family":"Nakamura","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shigeyuki","family":"Mukudai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renjie","family":"Bing","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael J.","family":"Garabedian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan C.","family":"Branski","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,11,24]]},"reference":[{"key":"77445_CR1","doi-asserted-by":"publisher","first-page":"82","DOI":"10.1016\/j.jvoice.2013.04.015","volume":"28","author":"N Govil","year":"2014","unstructured":"Govil, N., Paul, B. 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