ETHNOPHARMACOLOGICAL RELEVANCE Wu-Mei-Wan (WMW), a classic traditional Chinese herb medicine, is one of the most important formulations to treat digestive diseases from ancient times to the present. Previous study showed that WMW has satisfactory curative effects on experimental colitis, which motivating the application of WMW on colitis-associated complications. AIM OF THE STUDY Intestinal fibrosis is usually considered to be a common complication of inflammatory bowel disease (IBD), particularly Crohn's disease (CD). Currently, no effective preventive measures or medical therapies are available for that. This work was designed to evaluate the effect and related mechanism of WMW on chronic colitis-associated intestinal fibrosis mice model. MATERIALS AND METHODS The chronic colitis-associated intestinal fibrosis mice model was established by weekly intrarectal injection of 2,4,6-trinitrobenzene sulfonic acid (TNBS). The mice survival rate, disease activity index (DAI), colon length and histological score were examined to assess the therapeutic effect of WMW. Masson's trichrome staining, hydroxyproline assay, immunohistochemical staining and western blot analysis were used to evaluate fibrosis level. Colon inflammation was determined by ELISA and immunofluorescence staining. Immunofluorescence staining was used to evaluate fibroblasts proliferation and epithelial to mesenchymal transition (EMT), and the expression of key molecules in fibrosis was analyzed by western blot. RESULTS Here we showed that WMW alleviates chronic colitis with improved survival rate, DAI, colon length and histological score. WMW inhibited the progression of intestinal fibrosis, decreased the expression of various fibrosis markers, such as α-SMA, collagen I, MMP-9 and fibronectin. In addition, WMW treatment reduced cytokines IL-6 and IFN-γ, and downregulated proinflammatory NF-κBp65 and STAT3 signaling pathways. Importantly, administration of WMW led to the inhibition of colon fibroblast proliferation and EMT, which are important mediators during fibrosis. Several key profibrotic pathways, including TGF-β/Smad and Wnt/β-catenin pathways, were downregulated by WMW treatment. CONCLUSION Our work demonstrated that WMW can prevent intestinal fibrosis and the mechanisms involved may be related to the inhibition of colon fibroblasts activation.

中文翻译：

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Wu-Mei-Wan通过抑制成纤维细胞活化，改善了慢性结肠炎相关的肠道纤维化。

人种药理学吴美婉（WMW）是一种经典的传统中草药，是从古至今治疗消化系统疾病的最重要配方之一。先前的研究表明WMW对实验性结肠炎具有令人满意的疗效，这促使WMW在结肠炎相关并发症中的应用。研究目的肠纤维化通常被认为是炎症性肠病（IBD），尤其是克罗恩病（CD）的常见并发症。当前，尚无有效的预防措施或药物治疗。这项工作旨在评估WMW对慢性结肠炎相关性肠纤维化小鼠模型的影响及其相关机制。材料与方法通过每周直肠内注射2,4,6-三硝基苯磺酸（TNBS）建立慢性结肠炎相关性肠纤维化小鼠模型。检查小鼠存活率，疾病活动指数（DAI），结肠长度和组织学评分以评估WMW的治疗效果。使用Masson的三色染色，羟脯氨酸分析，免疫组织化学染色和蛋白质印迹分析来评估纤维化水平。通过ELISA和免疫荧光染色确定结肠炎症。免疫荧光染色用于评估成纤维细胞的增殖和上皮向间质转化（EMT），并通过蛋白质印迹分析纤维化中关键分子的表达。结果在这里，我们证明WMW可以缓解慢性结肠炎，并提高生存率，DAI，结肠长度和组织学评分。WMW抑制肠道纤维化的进程，降低各种纤维化标记物的表达，例如α-SMA，胶原I，MMP-9和纤连蛋白。此外，WMW治疗可减少细胞因子IL-6和IFN-γ，并下调促炎性NF-κBp65和STAT3信号通路。重要的是，WMW的使用导致结肠成纤维细胞增殖和EMT的抑制，这是纤维化过程中的重要介体。WMW处理下调了几个关键的纤维化途径，包括TGF-β/ Smad和Wnt /β-catenin途径。结论我们的工作证明WMW可以预防肠道纤维化，其参与的机制可能与抑制结肠成纤维细胞活化有关。降低了各种纤维化标记物的表达，例如α-SMA，胶原蛋白I，MMP-9和纤连蛋白。此外，WMW治疗可减少细胞因子IL-6和IFN-γ，并下调促炎性NF-κBp65和STAT3信号通路。重要的是，WMW的使用导致结肠成纤维细胞增殖和EMT的抑制，这是纤维化过程中的重要介体。WMW处理下调了几个关键的纤维化途径，包括TGF-β/ Smad和Wnt /β-catenin途径。结论我们的工作证明WMW可以预防肠道纤维化，其参与的机制可能与抑制结肠成纤维细胞活化有关。降低了各种纤维化标记物的表达，例如α-SMA，胶原蛋白I，MMP-9和纤连蛋白。此外，WMW治疗可减少细胞因子IL-6和IFN-γ，并下调促炎性NF-κBp65和STAT3信号通路。重要的是，WMW的使用导致结肠成纤维细胞增殖和EMT的抑制，这是纤维化过程中的重要介体。WMW处理下调了几个关键的纤维化途径，包括TGF-β/ Smad和Wnt /β-catenin途径。结论我们的工作证明WMW可以预防肠道纤维化，其参与的机制可能与抑制结肠成纤维细胞活化有关。并下调促炎性NF-κBp65和STAT3信号通路。重要的是，WMW的使用导致结肠成纤维细胞增殖和EMT的抑制，这是纤维化过程中的重要介体。WMW处理下调了几个关键的纤维化途径，包括TGF-β/ Smad和Wnt /β-catenin途径。结论我们的工作证明WMW可以预防肠道纤维化，其参与的机制可能与抑制结肠成纤维细胞活化有关。并下调促炎性NF-κBp65和STAT3信号通路。重要的是，WMW的使用导致结肠成纤维细胞增殖和EMT的抑制，这是纤维化过程中的重要介体。WMW处理下调了几个关键的纤维化途径，包括TGF-β/ Smad和Wnt /β-catenin途径。结论我们的工作证明WMW可以预防肠道纤维化，其参与的机制可能与抑制结肠成纤维细胞活化有关。