Department of Life, Health & Environmental Sciences, University of L'Aquila, Via Pompeo Spennati, Building Rita Levi Montalcini, Coppito, 67100 L'Aquila, Italy.
PhD School in Medicine and Public Health, Department of Life, Health and Environmental Sciences, University of L'Aquila, 67100 L'Aquila, Italy.
Int J Mol Sci. 2024 Aug 14;25(16):8862. doi: 10.3390/ijms25168862.
The mechanism underlying intestinal fibrosis, the main complication of inflammatory bowel disease (IBD), is not yet fully understood, and there is no therapy to prevent or reverse fibrosis. We evaluated, in in vitro cellular models, the ability of different classes of drugs currently used in IBD to counteract two pivotal processes of intestinal fibrosis, the differentiation of intestinal fibroblasts to activated myofibroblasts using CCD-18Co cells, and the epithelial-to-mesenchymal transition (EMT) of intestinal epithelial cells using Caco-2 cells (IEC), both being processes induced by transforming growth factor-β1 (TGF-β1). The drugs tested included mesalamine, azathioprine, methotrexate, prednisone, methylprednisolone, budesonide, infliximab, and adalimumab. The expression of fibrosis and EMT markers (collagen-I, α-SMA, pSmad2/3, occludin) was assessed by Western blot analysis and by immunofluorescence. Of the drugs used, only prednisone, methylprednisolone, budesonide, and adalimumab were able to antagonize the pro-fibrotic effects induced by TGF-β1 on CCD-18Co cells, reducing the fibrosis marker expression. Methylprednisolone, budesonide, and adalimumab were also able to significantly counteract the TGF-β1-induced EMT process on Caco-2 IEC by increasing occludin and decreasing α-SMA expression. This is the first study that evaluates, using in vitro cellular models, the direct antifibrotic effects of drugs currently used in IBD, highlighting which drugs have potential antifibrotic effects.
肠道纤维化的发病机制,即炎症性肠病(IBD)的主要并发症,尚未完全阐明,也没有预防或逆转纤维化的疗法。我们使用不同类别的药物在体外细胞模型中评估其预防和逆转纤维化的能力,这些药物目前用于治疗 IBD,包括 5-氨基水杨酸、硫唑嘌呤、甲氨蝶呤、泼尼松、甲泼尼龙、布地奈德、英夫利昔单抗和阿达木单抗。这些药物可拮抗转化生长因子-β1(TGF-β1)诱导的两种关键的肠道纤维化过程,即肠成纤维细胞向激活的肌成纤维细胞分化(使用 CCD-18Co 细胞)和肠道上皮细胞向间充质细胞转化(EMT)(使用 Caco-2 细胞)。纤维化和 EMT 标志物(I 型胶原、α-SMA、pSmad2/3、occludin)的表达通过 Western blot 分析和免疫荧光检测。在使用的药物中,只有泼尼松、甲泼尼龙、布地奈德和阿达木单抗能够拮抗 TGF-β1 对 CCD-18Co 细胞诱导的促纤维化作用,降低纤维化标志物的表达。甲泼尼龙、布地奈德和阿达木单抗还能够通过增加 occludin 表达和减少 α-SMA 表达,显著拮抗 TGF-β1 诱导的 Caco-2 IEC 的 EMT 过程。这是第一项使用体外细胞模型评估目前用于 IBD 的药物的直接抗纤维化作用的研究,突出了哪些药物具有潜在的抗纤维化作用。