Nifuroxazide ameliorates pulmonary fibrosis by blocking myofibroblast genesis: a drug repurposing study

• Published in: Respiratory research Vol. 23; no. 1; p. 32
• Main Authors: Gan, Cailing, Zhang, Qianyu, Liu, Hongyao, Wang, Guan, Wang, Liqun, Li, Yali, Tan, Zui, Yin, Wenya, Yao, Yuqin, Xie, Yongmei, Ouyang, Liang, Yu, Luoting, Ye, Tinghong
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 16-02-2022; BioMed Central; BMC
• Subjects: Antibodies; Anticancer properties; Assaying; Biocompatibility; Bleomycin; Cell migration; Cytotoxicity; Diabetes mellitus; Diabetic nephropathy; Diarrhea; Drug development; Drug therapy; Drugs; EMT; Enzyme-linked immunosorbent assay; Epithelial cells; Epithelium; Evaluation; Fibroblasts; Fibrosis; Flow cytometry; Genetic aspects; Growth factors; Health aspects; Hydroxyproline; Immune system; Immunofluorescence; Immunoglobulins; Immunohistochemistry; Immunosuppressive agents; In vivo methods and tests; Inflammatory diseases; IPF; Lung diseases; Mathematical analysis; Mesenchyme; Nephropathy; Nifuroxazide; Nitrofurantoin; Pathogenesis; Patient outcomes; Polyethylene glycol; Proteins; Pulmonary fibrosis; Side effects; Smad protein; Software; Stat3; Stat3 protein; Testing; TGF-β1/Smad; Toxicity; Transcription; Transforming growth factor-b1; Tumors; China; Beijing China; Grand Island New York; United States > US; Stat3; TGF-β1/Smad; EMT; Nifuroxazide; IPF
• ISSN: 1465-993X; 1465-9921; 1465-993X; 1465-9921
• DOI: 10.1186/s12931-022-01946-6

Idiopathic pulmonary fibrosis (IPF) is a serious interstitial lung disease with a complex pathogenesis and high mortality. The development of new drugs is time-consuming and laborious; therefore, research on the new use of old drugs can save time and clinical costs and even avoid serious side effects. Nifuroxazide (NIF) was originally used to treat diarrhoea, but more recently, it has been found to have additional pharmacological effects, such as anti-tumour effects and inhibition of inflammatory diseases related to diabetic nephropathy. However, there are no reports regarding its role in pulmonary fibrosis.
The therapeutic effect of NIF on pulmonary fibrosis in vivo was measured by ELISA, hydroxyproline content, H&E and Masson staining, immunohistochemistry (IHC) and western blot. Immune cell content in lung tissue was also analysed by flow cytometry. NIF cytotoxicity was evaluated in NIH/3T3 cells, human pulmonary fibroblasts (HPFs), A549 cells and rat primary lung fibroblasts (RPLFs) using the MTT assay. Finally, an in vitro cell model created by transforming growth factor-β1 (TGF-β1) stimulation was assessed using different experiments (immunofluorescence, western blot and wound migration assay) to evaluate the effects of NIF on the activation of NIH/3T3 and HPF cells and the epithelial-mesenchymal transition (EMT) and migration of A549 cells.
In vivo, intraperitoneal injection of NIF relieved and reversed pulmonary fibrosis caused by bleomycin (BLM) bronchial instillation. In addition, NIF inhibited the expression of a variety of cellular inflammatory factors and immune cells. Furthermore, NIF suppressed the activation of fibroblasts and EMT of epithelial cells induced by TGF-β1. Most importantly, we used an analytical docking experiment and thermal shift assay to further verify that NIF functions in conjunction with signal transducer and activator of transcription 3 (Stat3). Moreover, NIF inhibited the TGF-β/Smad pathway in vitro and decreased the expression of phosphorylated Stat3 in vitro and in vivo.
Taken together, we conclude that NIF inhibits and reverses pulmonary fibrosis, and these results support NIF as a viable therapeutic option for IPF treatment.