肠道纤维化的潜在机制,炎症性肠病(IBD)的主要并发症,还没有完全理解,并且没有预防或逆转纤维化的疗法。我们评估,在体外细胞模型中,目前用于IBD的不同类别的药物抵抗肠纤维化的两个关键过程的能力,使用CCD-18Co细胞将肠道成纤维细胞分化为活化的肌成纤维细胞,和使用Caco-2细胞(IEC)的肠上皮细胞的上皮-间质转化(EMT),都是转化生长因子-β1(TGF-β1)诱导的过程。测试的药物包括美沙拉嗪,硫唑嘌呤,甲氨蝶呤,泼尼松,甲基强的松龙,布地奈德,英夫利昔单抗,和阿达木单抗.纤维化和EMT标志物(胶原蛋白-I,α-SMA,pSmad2/3,occludin)通过蛋白质印迹分析和免疫荧光进行评估。在使用的药物中,只有泼尼松,甲基强的松龙,布地奈德,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。甲基强的松龙,布地奈德,和阿达木单抗还能够通过增加闭塞蛋白和降低α-SMA表达来显著抵消TGF-β1诱导的Caco-2IEC的EMT过程。这是第一项评估的研究,使用体外细胞模型,目前用于IBD的药物的直接抗纤维化作用,强调哪些药物具有潜在的抗纤维化作用。
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.