目的 运用网络药理学结合实验验证研究冯了性风湿跌打药酒治疗类风湿关节炎的作用机制。方法 借助前期研究中超高效液相色谱–四极杆–静电场轨道阱高分辨质谱联用技术分析冯了性风湿跌打药酒的活性成分，通过TCMSP数据库、Swiss Target Prediction数据库筛选活性成分靶点，Drugbank、OMIM、GeneCards、TTD、Disgenet数据库检索类风湿性关节炎的疾病靶点。利用STRING数据库绘制蛋白相互作用（PPI）网络图，分析冯了性风湿跌打药酒治疗类风湿关节炎的核心靶点。通过DAVID数据库进行基因本体（GO）功能注释和京都基因与基因组百科全书（KEGG）通路富集分析，预测其富集的信号通路并进行可视化展示。利用AutoDockTools 1.5.6软件进行分子对接。利用牛Ⅱ型胶原乳剂诱导建立类风湿关节炎大鼠模型，进行关节炎评分，组织病理学检查，PCR分析关键基因mRNA表达水平，ELISA法检测低氧诱导因子-1α（HIF-1α）和白细胞介素-6（IL-6）水平。结果 冯了性风湿跌打药酒中46个核心化学成分对应靶点661个，搜索收集获得疾病靶点2 604个，两者交集后获得283个冯了性风湿跌打药酒治疗类风湿关节炎的潜在靶点。分析得到蛋白激酶B1（Akt1）、肿瘤蛋白p53（TP53）、血管内皮生长因子A（VEGFA）等10个核心靶点，10条GO相关条目及15条KEGG通路，关键通路包括丝裂原活化蛋白激酶（MAPK）信号通路、磷脂酰肌醇-3-激酶（PI3K）/Akt信号通路、Ras信号通路、Rap1信号通路、HIF-1信号通路等。分子对接结果表明，秦皮乙素、5,7-二羟基香豆素、东莨菪苷核心成分与Akt1、HIF-1α核心靶点均具有良好的结合能力。动物实验结果显示，冯了性风湿跌打药酒可以缓解类风湿关节炎大鼠的关节肿度，降低关节指数，HE染色结果显示冯了性风湿跌打药酒减轻滑膜增生、软骨退化，PCR结果表明冯了性风湿跌打药酒显著降低HIF-1α mRNA水平，ELISA实验结果显示冯了性风湿跌打药酒显著降低IL-6、HIF-1α水平。结论 利用网络药理学分析，初步揭示了冯了性风湿跌打药酒可能作用于Akt1、PI3K等靶点干预类风湿关节炎的疾病进程，结合药效学实验，发现冯了性风湿跌打药酒可能是通过降低IL-6、HIF-1α，对类风湿关节炎起到治疗作用。

Objective To explore the material basis and potential mechanism of Fengliaoxing Fengshi Dieda Yaojiu in treatment of rheumatoid arthritis by network pharmacology and experimental verification. Methods Ultra-high performance liquid chromatography coupled with quadrupole/electrostatic field orbital trap high resolution mass spectrometry (UHPLC-Q Exactive Focus MS/MS) was developed to rapidly analyze and identify the chemical components in the Fengliaoxing Fengshi Dieda Yaojiu. The active components and targets of Fengliaoxing Fengshi Dieda Yaojiu and rheumatoid arthritis disease targets were obtained through network pharmacology-related database and analysis platform. The protein- protein (PPI) interaction map was drawn by STRING database and core targets of Fengliaoxing Fengshi Dieda Yaojiu in treatment of rheumatoid arthritis were analyzed. DAVID database was used to perform GO and KEGG enrichment analysis on the intersection targets. Molecular docking was performed using AutoDockTools 1.5.6 software. A rat model of rheumatoid arthritis induced by bovine typeⅡ collagen emulsion was established. The degree of arthritis was assessed by visual scoring. Histopathology examination to detect joint changes, PCR analysis to detect key protein mRNA expression level, ELISA to detect HIF-1α and IL-6 levels. Results A total of 46 active components were obtained by UHPLC-Q Exactive Focus MS/MS of Fengliaoxing Fengshi Dieda Yaojiu, which corresponded to 661 targets and 2 604 disease targets were screened. After the intersection of two targets, 283 potential targets of Fengliaoxing Fengshi Dieda Yaojiu in the treatment of rheumatoid arthritis were obtained. Ten core targets such as Akt1, TP53, and VEGFA were obtained by topological analysis. A total of 10 GO-related items and 15 KEGG pathways, mainly involving MAPK signaling pathway, PI3K/Akt signaling pathway, Ras signaling pathway, HIF-1 signaling pathway, Rap1 and other signaling pathways were gained by enrichment analysis. The results of molecular docking showed that the core components such as esculetin, 5,7-dihydroxycoumarin, scopolin had good binding ability to core targets such as Akt1 and HIF-1α. The results of animal experiments proved that Fengliaoxing Fengshi Dieda Yaojiu can alleviate joint swelling in the mice with rheumatoid arthritis. The results of ELISA showed that Fengliaoxing Fengshi Dieda Yaojiu lowered the levels of IL-6 and HIF-1α. PCR showed that Fengliaoxing Fengshi Dieda Yaojiu down-regulated the mRNA level of HIF-1α. Conclusion Through network pharmacological analysis, it was preliminatively revealed that Fengliaoxing Fengshi Dieda Yaojiu may improve the disease process of rheumatoid arthritis by acting on Akt1, PI3K, and other targets. Combined with pharmacodynamic experiments, it was found that Fengliaoxing Fengshi Dieda Yaojiu may play a therapeutic role in rheumatoid arthritis by reducing IL-6 and HIF-1α.

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国家自然科学基金面上项目（81973419）；陕西省重点研发计划一般项目（2022SF-315）；陕西省中医药研究院“苗圃计划”一搬项目（2021-26）