基于UPLC-MS/MS、网络药理学方法预测红芪免疫调节的核心靶点及作用通路，通过分子对接、 分子动力学技术对网络药理学结果进行验证，探讨红芪入血成分免疫调节的作用机制。方法基于UPLC-MS/ MS 技术定性定量红芪入血成分；通过TCMSP、本草组鉴数据库筛选红芪入血成分对应靶点；以DisGeNET、 OMIM、TTD、MalaCards 数据库获取免疫相关疾病靶点；构建“红芪入血成分-免疫相关疾病”网络；进行 GO、KEGG 富集分析，绘制PPI 网络；应用分子对接、分子动力学技术进行验证。结果UPLC-MS/MS 法共 鉴定8 个原型入血成分，协同作用于101 个靶点，参与免疫反应、基因表达的正调控、受体结合、细胞因子活 性等538 个生物学过程，涉及HIF-1、Toll 样受体、JAK-STAT、T 细胞受体、PI3K-Akt、FoxO 等116 条信号 通路。核心靶点为MAPK14、PTGS2、MMP9、PPARG、CCND1 等。分子对接结果显示，芒柄花素、毛蕊异黄 酮与MAPK14 的对接结合活性较高，分子动力学模拟进一步验证了芒柄花素、毛蕊异黄酮与MAPK14 的结合 具有较好的结构稳定性及结合亲和力。结论通过血清药物化学、网络药理学及分子动力学相互印证，揭示了 红芪调节免疫的物质基础及机制，可为其作用机制研究提供科学依据。

To predict the core targets and action pathways of Hedysari Radix based on UPLC-MS/MS and network pharmacology methods， and to verify the results of network pharmacology by molecular docking and molecular dynamics techniques. This article aims to investigate immune regulation mechanism of effective components absorbed into blood from Hedysari Radix. Methods Qualitative quantification of effective components absorbed into blood from Hedysari Radix were operated by using UPLC-MS/MS technique. The corresponding targets of effective components absorbed into blood from Hedysari Radix were screened by TCMSP and HERB databases. Targets of immune-related disease were obtained through DisGeNET，OMIM，TTD，and MalaCards databases. The network of “components absorbed into blood from Hedysari Radix-immune-related diseases”was then constructed. GO and KEGG enrichment analysis and mapped the PPI network were performed. Molecular docking and molecular dynamics techniques were applied for validation. Results A total of 8 prototype components absorbed into blood，synergistically acting on 101 targets，were identified by UPLC-MS/MS. They mediated 538 biological processes including immune response，positive regulation of gene expression，receptor binding，and cytokine activity. Meanuhile，116 signaling pathways， such as HIF-1， Toll-like receptor， JAK-STAT， T cell receptor， PI3K-Akt， and FoxO etc. were involved. The core targets were MAPK14， PTGS2， MMP9， PPARG， CCND1， etc. . The results of molecular docking showed that formononetin and calycosin had strong docking binding activity with MAPK14. And molecular dynamics simulations further demonstrated that the binding between MAPK14 and formononetin or calycosin had good structural stability and binding affinity. Conclusion The results of serum pharmacochemistry，network pharmacology and molecular dynamics were verified to reveal the material basis and mechanism of Hedysari Radix in regulating immunity. The aim of this study is to provide scientific basis for its immunomodulatory mechanism.

R285.5

国家自然科学基金地区基金项目（82160730，81860683）；甘肃省重点研发计划项目（21YF5FA133）；“ 双一流” 科研重点项目 （GSSYLXM-05）；敦煌医学与转化教育部重点实验室项目（DHYX20-12）。