Department of Colorectal Surgery, Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Suzhou, 215000, China.
Department of Gynaecology, Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Suzhou, 215000, China.
J Ethnopharmacol. 2023 Oct 28;315:116678. doi: 10.1016/j.jep.2023.116678. Epub 2023 May 30.
Xue-Jie-San (XJS), as a traditional Chinese herb prescription, has satisfactory effects on improving clinical symptoms and facilitating the healing of intestinal ulcers in patients with Crohn's disease (CD). This motivates the application of XJS on CD-associated complications.
Intestinal fibrosis is a debilitating complication of CD. Currently, there is no effective medication available for preventing or reversing CD-related intestinal fibrosis. This study aimed to assess the efficacy and underlying mechanisms of XJS in the treatment of colitis-associated intestinal fibrosis.
A rat model of CD-related intestinal fibrosis was induced by 2,4,6-trinitrobenzene sulfonic acid administration and treated with XJS. The pathological changes of intestinal fibrosis were evaluated using Masson staining. Collagen deposition and epithelial-to-mesenchymal transition (EMT) were verified by immunohistochemical staining and Western blot analysis. Endothelial-to-mesenchymal transition (EndoMT) was assessed with immunofluorescence and immunohistochemical staining as well as Western blot analysis. Transmission electron microscopy was utilized to observe autophagosomes. The levels of autophagy-related proteins were detected via immunofluorescence staining and Western blot. Finally, the mTOR/ULK1 signaling pathway regulated by Notch1 or FGL1 was analyzed by Western blot.
The results found that XJS ameliorated intestinal fibrosis through reducing the deposition of collagens such as Collagen 1 and Collagen 3. XJS inhibited the EMT process by increasing E-cadherin levels and decreasing the expressions of N-cadherin, Vimentin and Snail, which played a crucial role in collagen secretion and intestinal fibrosis. In addition, XJS also repressed the EndoMT process as reflected by the upregulation of CD31 and VE-cadherin levels and the downregulation of FSP1 and α-SMA expressions. Autophagy was activated following XJS treatment via suppression of the mTOR/ULK1 signaling pathway. Furthermore, XJS acted as an inhibitor of Notch1 and FGL1 signals, both of which regulated the mTOR signaling.
Our findings validated that XJS prevented the early development of CD-related intestinal fibrosis by blocking the Notch1 and FGL1 signaling pathways to activate autophagy and thereby inhibit EMT and EndoMT.
血竭散（XJS）作为一种中药方剂，对改善克罗恩病（CD）患者的临床症状和促进肠道溃疡愈合有满意的效果。这促使 XJS 应用于 CD 相关并发症。
肠道纤维化是 CD 的一种衰弱性并发症。目前，尚无有效的药物可用于预防或逆转 CD 相关的肠道纤维化。本研究旨在评估 XJS 治疗结肠炎相关肠道纤维化的疗效和潜在机制。
采用 2,4,6-三硝基苯磺酸（TNBS）诱导大鼠 CD 相关肠道纤维化模型，并给予 XJS 治疗。采用 Masson 染色评估肠道纤维化的病理变化。通过免疫组化染色和 Western blot 分析验证胶原沉积和上皮间质转化（EMT）。通过免疫荧光和免疫组化染色以及 Western blot 分析评估内皮间质转化（EndoMT）。采用透射电子显微镜观察自噬体。通过免疫荧光染色和 Western blot 检测自噬相关蛋白水平。最后，通过 Western blot 分析 Notch1 或 FGL1 调节的 mTOR/ULK1 信号通路。
结果发现，XJS 通过减少 Collagen 1 和 Collagen 3 等胶原的沉积来改善肠道纤维化。XJS 通过增加 E-cadherin 水平和降低 N-cadherin、Vimentin 和 Snail 的表达来抑制 EMT 过程，这对胶原分泌和肠道纤维化起着关键作用。此外，XJS 还通过上调 CD31 和 VE-cadherin 水平和下调 FSP1 和 α-SMA 表达来抑制 EndoMT 过程。XJS 通过抑制 mTOR/ULK1 信号通路来激活自噬。此外，XJS 作为 Notch1 和 FGL1 信号的抑制剂，这两种信号都调节 mTOR 信号。
我们的研究结果证实，XJS 通过阻断 Notch1 和 FGL1 信号通路激活自噬，从而抑制 EMT 和 EndoMT，防止 CD 相关肠道纤维化的早期发展。
