Exploring Combined Therapy: Disulfiram and Antifibrotic Agents in Attenuating Pulmonary Fibrosis in Bleomycin-Induced Mouse Model

<bold>Introduction:</bold> Pulmonary fibrosis, characterized by excessive accumulation of extracellular matrix, impairs lung function. Deficiencies in CCR2 and CCR5, and their ligands attenuate bleomycin (BLM)-induced lung fibrosis. Disulfiram, an anti-alcoholism drug, inhibits monocyte/macrophage migration by blocking FROUNT, an intracellular regulator of CCR2/CCR5 signaling. <bold>Methods:</bold> We investigated the antifibrotic effect of disulfiram and pirfenidone by conducting respiratory function tests and histopathological examinations in BLM-induced pulmonary fibrosis mouse model. Comprehensive gene expression analysis and RNA in situ hybridization to explore the therapeutic mechanisms. <bold>Results:</bold> Administration of disulfiram improved lung compliance of mice in lung function test, and reduced collagen deposition in the lung tissue, determined by Elastica Masson-Goldner staining. The sircol collagen assay demonstrated the reduction of the total collagen level in the disulfiram-administered lung tissue. The combined administration of disulfiram and pirfenidone synergistically augmented the antifibrotic effects in BLM-induced pulmonary fibrosis in mice. Comprehensive gene expression analysis and RNA in situ hybridization of murine lung tissues provided clues to elucidating the mechanisms underpinning inhibition of pulmonary fibrosis by disulfiram. <bold>Conclusions:</bold> Our study illuminates the effectiveness of disulfiram as a monotherapy for fibrosis and underscores its synergistic potential with pirfenidone. These findings provide a foundation for further research of disulfiram-based interventions and pave the way for the development of novel antifibrotic strategies.