乌梅丸通过调节 RAGE 信号通路抑制 IBD 的血管生成和炎症：网络药理学分析及实验证据。

Wumei Wan attenuates angiogenesis and inflammation by modulating RAGE signaling pathway in IBD: Network pharmacology analysis and experimental evidence.

机构信息

No. 1 Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing 210023, PR China.

Collaborative Innovation Center for Cancer Medicine, Jiangsu Province Hospital of Chinese Medicine, The Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, PR China.

出版信息

Phytomedicine. 2023 Mar;111:154658. doi: 10.1016/j.phymed.2023.154658. Epub 2023 Jan 12.

DOI:10.1016/j.phymed.2023.154658

PMID:36706698

Abstract

BACKGROUND

Wumei Wan (WMW) has been used to address digestive disorder for centuries in traditional Chinese medicine. Previous studies have demonstrated its anti-colitis efficacy, but the underlying mechanism of its action remains to be further clarified.

PURPOSE

To investigate the underlying mechanisms of WMW in the treatment of chronic ulcerative colitis (UC) through network pharmacology and experimental validation.

METHODS

Traditional Chinese Medicine Systems Pharmacology (TCMSP) platform were used to identify the ingredients and potential targets of WMW. The microarray gene data GSE75214 datasets from GEO database was used to define UC-associated targets. Cytoscape3.7.2 was employed to construct the protein-protein interaction (PPI) network and compounds-disease targets network. GO enrichment analysis and KEGG pathway analysis were performed by R software for functional annotation. UPLC-TOF-MS/MS method was used to quantitatively analyze the active ingredients of WMW. For experimental validation, three cycles of 2% dextran sulfate sodium salt (DSS) were used to construct chronic colitis model. The hub targets and signal pathway were detected by qPCR, ELISA, western blotting , immunohistochemical and immunofluorescence.

RESULTS

Through network analysis, 104 active ingredients were obtained from WMW, and 47 of these ingredients had potential targets for UC. A total of 41 potential targets of WMW and 13 hub targets were identified. KEGG analysis showed that WMW involved in advanced glycation end products-receptor of advanced glycation end products (AGE-RAGE) signaling pathway. Taxifolin, rutaecarpine, kaempferol, quercetin, and luteolin of WMW were the more highly predictive components related to the AGE-RAGE signaling pathway. In vivo validation, WMW improved DSS-induced colitis, reduced the expression of inflammatory cytokines and chemokines. Notably, it significantly decreased the mRNA expression of Spp1, Serpine1, Mmp2, Mmp9, Ptgs2, Nos2, Kdr and Icam1, which were associated with angiogenesis. In addition, we confirmed WMW inhibited RAGE expression and diminished DSS-induced epithelial barrier alterations CONCLUSION: Our results initially demonstrated the effective components and the strong anti-angiogenic activity of WMW in experimental chronic colitis. Sufficient evidence of the satisfactory anti-colitis action of WMW was verified in this study, suggesting its potential as a quite prospective agent for the therapy of UC.

摘要

背景

乌梅丸在中医药中已有数百年治疗消化系统疾病的历史。先前的研究已经证明了其抗结肠炎的疗效，但作用机制仍需进一步阐明。

目的

通过网络药理学和实验验证研究乌梅丸治疗慢性溃疡性结肠炎（UC）的潜在机制。

方法

利用中药系统药理学（TCMSP）平台识别乌梅丸的成分和潜在靶点。从 GEO 数据库的 GSE75214 数据集确定与 UC 相关的靶点。采用 Cytoscape3.7.2 构建蛋白质-蛋白质相互作用（PPI）网络和化合物-疾病靶点网络。使用 R 软件进行 GO 富集分析和 KEGG 通路分析进行功能注释。采用 UPLC-TOF-MS/MS 方法定量分析乌梅丸的活性成分。实验验证采用 3 周期 2%葡聚糖硫酸钠（DSS）构建慢性结肠炎模型。通过 qPCR、ELISA、western blot、免疫组化和免疫荧光检测核心靶点和信号通路。

结果

通过网络分析，从乌梅丸中获得 104 种活性成分，其中 47 种成分对 UC 有潜在靶点。确定了乌梅丸的 41 个潜在靶点和 13 个核心靶点。KEGG 分析表明，乌梅丸参与晚期糖基化终产物-受体的高级糖基化终产物（AGE-RAGE）信号通路。乌梅丸中的杨梅素、荷叶碱、山奈酚、槲皮素、木犀草素等成分与 AGE-RAGE 信号通路相关性更高。体内验证结果表明，乌梅丸可改善 DSS 诱导的结肠炎，降低炎症细胞因子和趋化因子的表达。值得注意的是，它显著降低了与血管生成相关的 Spp1、Serpine1、Mmp2、Mmp9、Ptgs2、Nos2、Kdr 和 Icam1 的 mRNA 表达。此外，我们证实乌梅丸抑制了 RAGE 的表达，减轻了 DSS 诱导的上皮屏障改变。