目的 基于体外模拟消化、发酵和网络药理学探讨桑叶酚类物质对于阿尔茨海默病（Alzheimer’s disease，AD）、癌症和糖尿病3种疾病的潜在共同作用靶点，为研究桑叶治疗多种疾病提供理论基础。方法 新鲜桑叶冷冻干燥粉碎后模拟体外消化及肠道微生物发酵过程，取不同消化（或发酵）阶段产物，利用UPLC-Q-TOF-MS/MS技术分析其中的特征酚类物质，结合Swiss Target Prediction和Genecards数据库筛选特征酚类物质治疗AD、癌症和糖尿病3种疾病的潜在作用靶点，采用String平台构建蛋白相互作用（protein-protein interaction，PPI）网络图，再用Cytoscape软件分析特征酚类物质与以上3种疾病作用的共同核心靶点，并筛选出其中的主要活性组分；通过David数据库对共同核心靶点进行基因本体（gene ontology，GO）功能和京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）信号通路富集分析；使用Autodock vina和PyMol软件对主要活性组分和共同核心靶点进行分子对接。结果 桑叶体外消化（或发酵）各阶段共检测到19种特征酚类物质，其中主要活性组分为咖啡酸、槲皮素和山柰酚。特征酚类物质与3种疾病交集靶点的共同靶点有48种。PPI得出特征酚类物质对3种疾病作用的10个共同核心靶点（AKT1、TNF、EGFR、PTGS2、SRC、TLR4、CASP3、ESR1、STAT3和MMP9）。对核心靶点进行富集分析，得到120个生物过程、11个细胞组分、14个分子功能和35条信号通路，其中炎症相关通路占比最高。主要活性组分与核心靶点进行分子对接模拟后，结合能均小于−5 kcal/mol。结论 桑叶不同体外消化（发酵）过程中产生的特征酚类物质可能通过抑制慢性炎症反应对AD、癌症和糖尿病3种疾病起到治疗效应。

Objective To explore the potential synergistic targets of phenols from Sangye (Mori Folium) for Alzheimer’s disease (AD), cancer and diabetes mellitus based on the in vitro simulated digestion, fermentation and network pharmacology, to provid the theoretical basis in the treatment of various diseases with Mori Folium. Methods After freeze-dried and pulverized, the fresh Mori Folium were used for in vitro digestion and intestinal microbial fermentation process. Different products from digestion (or fermentation) stage were analyzed to screen out the characteristic phenolic substances by UPLC-Q-TOF-MS/MS. The Swiss Target Prediction and Genecards database were used to screen the potential targets of characteristic phenolic substances in the treatment of AD, cancer and diabetes mellitus. The String platform was used to establish a protein-protein interaction (PPI) network diagram. Then the common key targets between the characteristic phenolic substances and the three diseases were analyzed by Cytoscape, and the main active components were screened out. The gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) signaling pathway enrichment analysis of the common core targets were performed on the David database. Finally, the molecular docking of the main active components and common core targets was performed via Autodock vina and PyMol. Results A total of 19 characteristic phenolic substances were detected in in vitro digestion (or fermentation) stage of Mori Folium, and the main active components were caffeic acid, quercetin and kaempferol. There were 48 common targets among the characteristic phenolic substances and the three diseases. Ten common core targets (AKT1, TNF, EGFR, PTGS2, SRC, TLR4, CASP3, ESR1, STAT3 and MMP9) between the characteristic phenolic substances and the three diseases were obtained from PPI analysis. After enrichment analysis of common core targets were performed, then 120 biological processes, 11 cell components, 14 molecular functions and 35 signaling pathways were screened out, in which inflammation-related pathways accounted for the highest proportion. The binding energy between the main active components and common core targets were lower than −5 kcal/mol during the molecular docking. Conclusion The characteristic phenolic substances produced during different in vitro digestion (fermentation) of Mori Folium may exert therapeutic effects on AD, cancer and diabetes, which may be mediated by inhibiting chronic inflammation.

R284;R285

江西省技术创新引导类计划项目（20212BDH81028）