基于UPLC-Orbitrap Fusion Lumos Tribrid-MS和网络药理学的正骨紫金丸化学成分及作用机制研究

UPLC-Orbitrap Fusion Lumis Tribrid-MS combined with network pharmacology and experimental validation to explore the active ingredients and mechanism of action of Zhenggu Zijin Wan

关键词:  正骨紫金丸  UPLC-Orbitrap Fusion Lumos Tribrid-MS  成分分析  网络药理学  活血化瘀

Key words:  Zhenggu Zijin Pill  UPLC-Orbitrap Fusion Lumos Tribrid-MS  Component analysis  Network pharmacology  Promoting blood circulation and resolving blood stasis

摘要:

目的 基于UPLC-Orbitrap Fusion Lumos Tribrid-MS技术联合网络药理学方法与实验验证预测正骨紫金丸活血化瘀的活性成分及作用机制。方法 首先，采用UPLC-Orbitrap Fusion Lumos Tribrid-MS技术快速表征正骨紫金丸中的化学成分。其次，通过网络药理学的研究方法构建“药物-成分-靶点”网络，获取关键靶点及主要活性成分，结合String平台与Cytoscape软件构建蛋白互作(PPI)网络，通过Matescape数据库富集分析通路，利用Discovery Studio 4.5软件进行分子对接验证。最后，建立急性软组织损伤动物模型，以急性软组织损伤评分与全血黏度为药效学指标开展药效学研究。结果 正骨紫金丸中共鉴定出包括黄酮类、生物碱类、有机酸类和香豆素类等化合物67个，其中大黄素、藁本内酯、肉桂酸、水杨酸、芦荟大黄素可能为正骨紫金丸活血化瘀的主要活性成分。蛋白互作网络拓扑分析得到TNF、ALB、AKT1等 26个核心靶点，KEGG 富集分析表明正骨紫金丸主要通过调控TNF、PI3K-Akt、NF-κB等信号通路发挥活血化瘀作用，分子对接结果显示正骨紫金丸主要活性成分与关键靶点结合良好，药效学结果表明正骨紫金丸可显著降低急性软组织损伤大鼠的全血粘度。结论 本研究明确了正骨紫金丸活血化瘀的活性成分和作用机制，同时表明其可能通过作用于多靶点、多通路整体调节，共同发挥活血化瘀作用，为其后续深入研究提供参考。

Abstract:

ABSTRACT: OBJECTIVE Based on UPLC-Orbitrap Fusion Lumis Tribrid-MS technology combined with network pharmacology methods, the active ingredients and mechanism of promoting blood circulation and resolving blood stasis in Zhenggu Zijin Pills are predicted. METHODS Using UPLC-Orbitrap Fusion Lumos Tribrid-MS technology to quickly characterize the chemical components in Zhenggu Zijin Pills, screening potential targets of Zhenggu Zijin Pills through databases such as Traditional Chinese Medicine System Pharmacological Analysis Platform (TCMSP) and Swiss Target Prediction, and searching for blood stasis related disease targets using GeneCards, Human Mendelian Genetic Database (OMIM), DrugBank, etc, Combining the String platform with Cytoscape software to construct protein interaction (PPI) networks; Enrichment and analysis of pathways through the Matescape database, molecular docking using Discovery Studio 4.5 software, and validation in animal experiments. RESULTS A total of 67 compounds including flavonoids, alkaloids, organic acids, and coumarins were identified in Zhenggu Zijin Wan. Among them, emodin, ligustilide, cinnamic acid, salicylic acid, aloe emodin, and others may be the main active ingredients for promoting blood circulation and removing blood stasis in Zhenggu Zijin Wan. Protein interaction network analysis revealed 26 core targets such as TNF, ALB, AKT1. KEGG enrichment analysis showed that Zhenggu Zijin Wan mainly regulates TNF, PI3K-Akt, NF- κ The B signaling pathway plays a role in promoting blood circulation and resolving stasis, and molecular docking results show that the main active ingredients of Zhenggu Zijin Pill bind well with key targets. CONCLUSION The study preliminarily clarifies the active ingredients and mechanism of action of Zhenggu Zijin Pill in promoting blood circulation and resolving blood stasis, and indicates that it may exert the effect of promoting blood circulation and resolving blood stasis together by acting on multiple targets and pathways, providing reference for its future in-depth research.