背景:DIREN是一种SHE民族医学,止血,清热,去除毒素的影响。它在临床上用于治疗消化道出血,如溃疡性结肠炎(UC)。
目的:纤维化是UC进展的病理改变之一,这使得对治疗的反应变得具有挑战性。我们旨在阐明DIREN在DSS诱导的UC中的作用,并试图从两个角度揭示其相关机制:肠道炎症和胶原蛋白沉积。
方法:使用2.5%葡聚糖硫酸钠(DSS)水溶液诱导小鼠结肠炎。使用疾病活动指数评估DIREN的治疗效果,组织病理学评分,和结肠长度。采用Masson和SiriusRed染色观察结肠的纤维化情况。TUNEL免疫荧光染色观察结肠上皮细胞凋亡。RNA-seq观察到差异基因和富集途径。采用免疫组织化学和RT-qPCR检测结肠组织中纤维化相关分子和局灶性粘附信号的表达。
结果:DIREN的给药导致UC小鼠的疾病活动指数(DAI)降低,同时促进结肠长度增加。DIREN减轻UC小鼠结肠杯状细胞的损失并维持肠粘膜屏障的完整性。Masson染色显示DIREN治疗结肠纤维化减少,天狼星红染色显示胶原蛋白Ⅰ沉积减少。DIREN降低结肠上皮细胞凋亡和基因表达,如CDH2、ITGA1和TGF-β2。此外,结肠组织转录组的GSEA分析结果表明,差异表达基因富集在粘着斑通路中。发现DIREN下调BAX的蛋白质表达,N-钙黏着蛋白,β-连环蛋白,整合素A1和Vinculin同时上调BCL2的蛋白表达。此外,它导致N-钙黏着蛋白和α-SMA的共表达。
结论:DIREN通过调节局灶性粘附和WNT/β-catenin信号通路改善结肠纤维化,对DSS诱导的UC具有保护作用。从而抑制成纤维细胞迁移并减少胶原蛋白分泌。
BACKGROUND: DIREN is a SHE ethnic medicine with stasis-resolving, hemostasis, clearing heat, and removing toxin effects. It is clinically used in the treatment of gastrointestinal bleeding, such as ulcerative colitis (UC).
OBJECTIVE: Fibrosis is one of the pathological changes in the progression of UC, which can make it challenging to respond to a treatment. We aimed to illuminate the role of DIREN in DSS-induced UC and tried to unveil its related mechanisms from two perspectives: intestinal inflammation and collagen deposition.
METHODS: A 2.5 % dextran sulfate sodium (DSS) water solution was used to induce colitis in mice. The therapeutic effect of DIREN was assessed using the disease activity index, histopathological score, and colon length. Masson and Sirius Red staining was used to observe the fibrosis in the colon. Apoptosis of colonic epithelial cells was observed by TUNEL immunofluorescence staining. RNA-seq observed differential genes and enrichment pathways. Immunohistochemistry and RT-qPCR were used to detect the expression of molecules related to fibrosis and focal adhesion signaling in colon tissue.
RESULTS: The administration of DIREN resulted in a reduction of disease activity index (DAI) in mice with UC while simultaneously promoting an increase in colon length. DIREN mitigated the loss of goblet cells in the colon of UC mice and maintained the integrity of the intestinal mucosa barrier. Masson staining revealed a reduction in colonic fibrosis with DIREN treatment, while Sirius red staining demonstrated a decrease in collagen Ⅰ deposition. DIREN reduced apoptosis of colonic epithelial cells and the expression of genes, such as CDH2, ITGA1, and TGF-β2. Additionally, the results of GSEA analysis of colon tissue transcriptome showed that the differentially expressed genes were enriched in the focal adhesion pathway. DIREN was found to downregulate the protein expression of BAX, N-cadherin, β-catenin, Integrin A1, and Vinculin while upregulating the protein expression of BCL2. Additionally, it led to the co-expression of N-cadherin and α-SMA.
CONCLUSIONS: DIREN exerts a protective effect against DSS-induced UC by ameliorating colonic fibrosis via regulation of focal adhesion and the WNT/β-catenin signaling pathway, thereby inhibiting fibroblast migration and reducing collagen secretion.