基于UPLC-MS/MS 和网络药理学、分子动力学探讨红芪免疫调节机制

目的 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...

Exploration the Immune Regulatory Mechanism of Hedysari Radix Based on Network Pharmacology，Molecular Dynamics，and UPLC-MS/MS

To predict the core targets and action pathways of Hedysari Radix based on UPLC-MS/MSand network pharmacology methods， and to verify the results of network pharmacology by molecular docking andmolecular dynamics techniques. This article aims to investigate immune regulation mechanism of effective componentsabsorbed into blood from Hedysari Radix. Methods Qualitative quantification of effective components absorbed intoblood from Hedysari Radix were operated by using UPLC-MS/MS technique. The corresponding targets of effectivecomponents absorbed into blood from Hedysari Radix were screened by TCMSP and HERB databases. Targets ofimmune-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 andKEGG enrichment analysis and mapped the PPI network were performed. Molecular docking and molecular dynamicstechniques 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 immuneresponse，positive regulation of gene expression，receptor binding，and cytokine activity. Meanuhile，116 signalingpathways， such as HIF-1， Toll-like receptor， JAK-STAT， T cell receptor， PI3K-Akt， and FoxO etc. wereinvolved. The core targets were MAPK14， PTGS2， MMP9， PPARG， CCND1， etc. . The results of moleculardocking showed that formononetin and calycosin had strong docking binding activity with MAPK14. And moleculardynamics simulations further demonstrated that the binding between MAPK14 and formononetin or calycosin had goodstructural stability and binding affinity. Conclusion The results of serum pharmacochemistry，network pharmacologyand molecular dynamics were verified to reveal the material basis and mechanism of Hedysari Radix in regulatingimmunity. The aim of this study is to provide scientific basis for its immunomodulatory mechanism.