目的 運(yùn)用網(wǎng)絡(luò)藥理學(xué)結(jié)合實(shí)驗(yàn)驗(yàn)證研究馮了性風(fēng)濕跌打藥酒治療類風(fēng)濕關(guān)節(jié)炎的作用機(jī)制。方法 借助前期研究中超高效液相色譜–四極桿–靜電場軌道阱高分辨質(zhì)譜聯(lián)用技術(shù)分析馮了性風(fēng)濕跌打藥酒的活性成分，通過TCMSP數(shù)據(jù)庫、Swiss Target Prediction數(shù)據(jù)庫篩選活性成分靶點(diǎn)，Drugbank、OMIM、GeneCards、TTD、Disgenet數(shù)據(jù)庫檢索類風(fēng)濕性關(guān)節(jié)炎的疾病靶點(diǎn)。利用STRING數(shù)據(jù)庫繪制蛋白相互作用（PPI）網(wǎng)絡(luò)圖，分析馮了性風(fēng)濕跌打藥酒治療類風(fēng)濕關(guān)節(jié)炎的核心靶點(diǎn)。通過DAVID數(shù)據(jù)庫進(jìn)行基因本體（GO）功能注釋和京都基因與基因組百科全書（KEGG）通路富集分析，預(yù)測其富集的信號通路并進(jìn)行可視化展示。利用AutoDockTools 1.5.6軟件進(jìn)行分子對接。利用牛Ⅱ型膠原乳劑誘導(dǎo)建立類風(fēng)濕關(guān)節(jié)炎大鼠模型，進(jìn)行關(guān)節(jié)炎評分，組織病理學(xué)檢查，PCR分析關(guān)鍵基因mRNA表達(dá)水平，ELISA法檢測低氧誘導(dǎo)因子-1α（HIF-1α）和白細(xì)胞介素-6（IL-6）水平。結(jié)果 馮了性風(fēng)濕跌打藥酒中46個核心化學(xué)成分對應(yīng)靶點(diǎn)661個，搜索收集獲得疾病靶點(diǎn)2 604個，兩者交集后獲得283個馮了性風(fēng)濕跌打藥酒治療類風(fēng)濕關(guān)節(jié)炎的潛在靶點(diǎn)。分析得到蛋白激酶B1（Akt1）、腫瘤蛋白p53（TP53）、血管內(nèi)皮生長因子A（VEGFA）等10個核心靶點(diǎn)，10條GO相關(guān)條目及15條KEGG通路，關(guān)鍵通路包括絲裂原活化蛋白激酶（MAPK）信號通路、磷脂酰肌醇-3-激酶（PI3K）/Akt信號通路、Ras信號通路、Rap1信號通路、HIF-1信號通路等。分子對接結(jié)果表明，秦皮乙素、5,7-二羥基香豆素、東莨菪苷核心成分與Akt1、HIF-1α核心靶點(diǎn)均具有良好的結(jié)合能力。動物實(shí)驗(yàn)結(jié)果顯示，馮了性風(fēng)濕跌打藥酒可以緩解類風(fēng)濕關(guān)節(jié)炎大鼠的關(guān)節(jié)腫度，降低關(guān)節(jié)指數(shù)，HE染色結(jié)果顯示馮了性風(fēng)濕跌打藥酒減輕滑膜增生、軟骨退化，PCR結(jié)果表明馮了性風(fēng)濕跌打藥酒顯著降低HIF-1α mRNA水平，ELISA實(shí)驗(yàn)結(jié)果顯示馮了性風(fēng)濕跌打藥酒顯著降低IL-6、HIF-1α水平。結(jié)論 利用網(wǎng)絡(luò)藥理學(xué)分析，初步揭示了馮了性風(fēng)濕跌打藥酒可能作用于Akt1、PI3K等靶點(diǎn)干預(yù)類風(fēng)濕關(guān)節(jié)炎的疾病進(jìn)程，結(jié)合藥效學(xué)實(shí)驗(yàn)，發(fā)現(xiàn)馮了性風(fēng)濕跌打藥酒可能是通過降低IL-6、HIF-1α，對類風(fēng)濕關(guān)節(jié)炎起到治療作用。

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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國家自然科學(xué)基金面上項(xiàng)目（81973419）；陜西省重點(diǎn)研發(fā)計劃一般項(xiàng)目（2022SF-315）；陜西省中醫(yī)藥研究院“苗圃計劃”一搬項(xiàng)目（2021-26）