Disulfiram Exerts Antifibrotic and Anti-Inflammatory Therapeutic Effects on Perimysial Orbital Fibroblasts in Graves' Orbitopathy

• Published in: International journal of molecular sciences 2022-05, Vol.23 (9), p.5261
• Main Authors: Wang, Xing, Ye, Huijing, Yang, Shenglan, Sha, Xiaotong, Wang, Xiandai, Zhang, Te, Chen, Rongxin, Xiao, Wei, Yang, Huasheng
• Format: Article
• Language: English
• Subjects: Anti-Inflammatory Agents - pharmacology; Cell division; Cell growth; Cells, Cultured; Chemokines; Cholecystokinin; Collagen; Collagen (type I); Connective tissue; Connective tissue growth factor; Coronaviruses; COVID-19; Disulfiram; Disulfiram - metabolism; Disulfiram - pharmacology; Drug dosages; Early intervention; Edema; Extracellular matrix; Fibroblasts; Fibroblasts - metabolism; Fibrosis; Graves Ophthalmopathy - drug therapy; Graves Ophthalmopathy - metabolism; Graves’ orbitopathy; Humans; Inflammation; Markers; Medical prognosis; Molecular modelling; Motility; mRNA; Muscles; myofibroblasts; Oculomotor system; perimysial orbital fibroblasts; Proteins; Severe acute respiratory syndrome coronavirus 2; Signal transduction; Transforming Growth Factor beta1 - metabolism; Transforming growth factor-b1
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23095261

Fibrosis of extraocular muscles (EOMs) is a marker of end-stage in Graves' orbitopathy (GO). To determine the antifibrotic and anti-inflammatory therapeutic effects and the underlying molecular mechanisms of disulfiram (DSF) on perimysial orbital fibroblasts (pOFs) in a GO model in vitro, primary cultures of pOFs from eight patients with GO and six subjects without GO (NG) were established. CCK-8 and EdU assays, IF, qPCR, WB, three-dimensional collagen gel contraction assays, cell scratch experiments, and ELISAs were performed. After TGF-β1 stimulation of pOFs, the proliferation rate of the GO group but not the NG group increased significantly. DSF dose-dependently inhibited the proliferation, contraction, and migration of pOFs in the GO group. Additionally, DSF dose-dependently inhibited fibrosis and extracellular matrix production markers (FN1, COL1A1, α-SMA, CTGF) at the mRNA and protein levels. Furthermore, DSF mediates antifibrotic effects on GO pOFs partially through the ERK-Snail signaling pathway. In addition, DSF attenuated HA production and suppressed inflammatory chemokine molecule expression induced by TGF-β1 in GO pOFs. In this in vitro study, we demonstrate the inhibitory effect of DSF on pOFs fibrosis in GO, HA production, and inflammation. DSF may be a potential drug candidate for preventing and treating tissue fibrosis in GO.