Mulberroside A is a polyhydroxylated stilbene active component of Morus alba L. Studies have shown that it has antitussive, antiasthmatic, tyrosinase and antioxidation activities. However, little is known about the metabolism of it in vitro and in vivo. In our study, an integrated strategy on the basis of UHPLC-Q-Exactive Plus Orbitrap MS and network pharmacology was established to comprehensively research the metabolic characteristic of mulberroside A for the first time. Plasma, urine, feces and liver tissues of rats in the blank group and drug group were collected after intragastric administration of mulberroside A at a dose of 150 mg/kg, and rat liver microsomes were cultured for in vitro metabolism experiment. The biological samples were processed by different methods and analyzed in positive and negative ion modes using UHPLC-Q-Exactive Plus Orbitrap MS. A total of 72 metabolites were finally identified based on the accurate molecular mass, retention time, MS/MS spectra and related literatures combined with the Compound Discoverer 3.1. The metabolic pathways were mainly hydrolysis, glucuronidation, hydrogenation, sulfation, hydroxylation, methylation and their composite reactions. In addition, a network pharmacology method was used to predict the mechanism of action of mulberroside A and its metabolites. In the end, 7 metabolites with high gastrointestinal absorption and drug-likeness and 167 targets were screened by Swiss ADME and Swiss Target Prediction. 1702 items of GO analysis and 158 related signaling pathways of KEGG were enriched using Metascape. This study established a novel integrated strategy based on UHPLC-Q-Exactive Plus Orbitrap MS and network pharmacology, which could systematically analyze the metabolism behavior of mulberroside A in vivo and in vitro of rats and provide basis for the further research of mulberroside A.

Mulberroside A 是一种多羟基茋活性成分桑白皮L. 研究表明，它具有镇咳、平喘、酪氨酸酶和抗氧化活性。然而，人们对它的新陈代谢知之甚少体外和体内. 本研究首次建立了基于UHPLC-Q-Exactive Plus Orbitrap MS和网络药理学的综合策略，全面研究桑椹苷A的代谢特性。灌胃桑椹苷A 150 mg/kg后，采集空白组和药物组大鼠血浆、尿液、粪便和肝组织，培养大鼠肝微粒体体外新陈代谢实验。通过不同的方法处理生物样品，并使用 UHPLC-Q-Exactive Plus Orbitrap MS 在正离子和负离子模式下进行分析。根据准确的分子质量、保留时间、MS/MS 谱图和相关文献，结合 Compound Discoverer 3.1，最终鉴定出 72 种代谢物。代谢途径主要为水解、葡萄糖醛酸化、氢化、硫酸化、羟基化、甲基化及其复合反应。此外，采用网络药理学方法预测桑椹苷A及其代谢物的作用机制。最终通过Swiss ADME和Swiss Target Prediction筛选出7个具有高胃肠道吸收和药物相似性的代谢物和167个靶点。使用 Metascape 富集了 1702 条 GO 分析项目和 158 条 KEGG 相关信号通路。本研究建立了一种基于 UHPLC-Q-Exactive Plus Orbitrap MS 和网络药理学的新型整合策略，可系统分析桑椹苷 A 的代谢行为。体内和体外并为桑椹苷A的进一步研究提供依据。