Chen Xu-Yang, Dong Yan-Chao, Yu Yuan-Yuan, Jiang Man, Bu Wen-Jie, Li Ping, Sun Zhi-Jie, Dong De-Li
Department of Pharmacology (The State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Key Laboratory of Cardiovascular Research, Ministry of Education), College of Pharmacy, Harbin Medical University, Harbin, China.
Department of Pharmacology, China Pharmaceutical University, Nanjing, China.
Br J Pharmacol. 2023 Dec;180(23):3008-3023. doi: 10.1111/bph.16190. Epub 2023 Aug 2.
Nitazoxanide is a therapeutic anthelmintic drug. Our previous studies found that nitazoxanide and its metabolite tizoxanide activated adenosine 5'-monophosphate-activated protein kinase (AMPK) and inhibited signal transducer and activator of transcription 3 (STAT3) signals. As AMPK activation and/or STAT3 inhibition are targets for treating pulmonary fibrosis, we hypothesized that nitazoxanide would be effective in experimental pulmonary fibrosis.
The mitochondrial oxygen consumption rate of cells was measured by using the high-resolution respirometry system Oxygraph-2K. The mitochondrial membrane potential of cells was evaluated by tetramethyl rhodamine methyl ester (TMRM) staining. The target protein levels were measured by using western blotting. The mice pulmonary fibrosis model was established through intratracheal instillation of bleomycin. The examination of the lung tissues changes were carried out using haematoxylin and eosin (H&E), and Masson staining.
Nitazoxanide and tizoxanide activated AMPK and inhibited STAT3 signalling in human lung fibroblast cells (MRC-5 cells). Nitazoxanide and tizoxanide inhibited transforming growth factor-β1 (TGF-β1)-induced proliferation and migration of MRC-5 cells, collagen-I and α-smooth muscle cell actin (α-SMA) expression, and collagen-I secretion from MRC-5 cells. Nitazoxanide and tizoxanide inhibited epithelial-mesenchymal transition (EMT) and inhibited TGF-β1-induced Smad2/3 activation in mouse lung epithelial cells (MLE-12 cells). Oral administration of nitazoxanide reduced the bleomycin-induced mice pulmonary fibrosis and, in the established bleomycin-induced mice, pulmonary fibrosis. Delayed nitazoxanide treatment attenuated the fibrosis progression.
Nitazoxanide improves the bleomycin-induced pulmonary fibrosis in mice, suggesting a potential application of nitazoxanide for pulmonary fibrosis treatment in the clinic.
硝唑尼特是一种治疗性驱虫药物。我们之前的研究发现，硝唑尼特及其代谢产物替唑尼特可激活5'-单磷酸腺苷激活蛋白激酶（AMPK）并抑制信号转导子和转录激活子3（STAT3）信号。由于AMPK激活和/或STAT3抑制是治疗肺纤维化的靶点，我们推测硝唑尼特对实验性肺纤维化有效。
使用高分辨率呼吸测定系统Oxygraph-2K测量细胞的线粒体氧消耗率。通过四甲基罗丹明甲酯（TMRM）染色评估细胞的线粒体膜电位。使用蛋白质免疫印迹法测量目标蛋白水平。通过气管内注入博来霉素建立小鼠肺纤维化模型。使用苏木精和伊红（H&E）以及Masson染色对肺组织变化进行检查。
硝唑尼特和替唑尼特在人肺成纤维细胞（MRC-5细胞）中激活AMPK并抑制STAT3信号。硝唑尼特和替唑尼特抑制转化生长因子-β1（TGF-β1）诱导的MRC-5细胞增殖和迁移、I型胶原蛋白和α-平滑肌肌动蛋白（α-SMA）表达以及MRC-5细胞分泌I型胶原蛋白。硝唑尼特和替唑尼特抑制上皮-间质转化（EMT）并抑制TGF-β1诱导的小鼠肺上皮细胞（MLE-12细胞）中Smad2/3激活。口服硝唑尼特可减轻博来霉素诱导的小鼠肺纤维化，并且在已建立的博来霉素诱导的小鼠中减轻肺纤维化。延迟给予硝唑尼特治疗可减弱纤维化进展。
硝唑尼特可改善博来霉素诱导的小鼠肺纤维化，提示硝唑尼特在临床上治疗肺纤维化具有潜在应用价值。
