目的 通过网络药理学、分子对接及实验验证探究秦艽Gentiana straminea中环烯醚萜苷抗类风湿性关节炎（rheumatoid arthritis，RA）的主要成分和作用机制。方法 通过文献报道和TCMSP数据收集并通过SwissADME数据库筛选环烯醚萜苷类成分，Pharmmaper数据库预测药物靶点；通过OMIM、GeneCards和TTD等数据库收集RA相关靶点，并通过Venn分析筛选出环烯醚萜苷抗RA的潜在作用靶点，通过构建蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络筛选环烯醚萜苷抗RA的核心靶点，通过基因本体（gene ontology，GO）功能及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析预测环烯醚萜苷抗RA的作用机制。以核心靶点为受体进行分子对接和分子动力学模拟筛选出环烯醚萜苷的关键成分，通过人风湿关节炎成纤维滑膜细胞（human fibroblast-like synoviocytes in rheumatoid arthritis，HFLS-RA）进行体外实验验证。结果 共收集获得18个具有类药性的环烯醚萜苷类成分、300个药物相关靶点和1 641个RA相关靶点；Venn分析筛选出103个环烯醚萜苷抗RA的潜在作用靶点；PPI网络分析发现原癌基因酪氨酸蛋白激酶Src（proto-oncogene tyrosine-protein kinase Src，SRC）、丝裂原活化蛋白激酶1（mitogen-activated protein kinase-1，MAPK1）、非受体酪氨酸激酶淋巴细胞特异性蛋白酪氨酸激酶（non-receptor tyrosine kinase lymphocyte cell-specific protein-tyrosine kinase，LCK）、表皮生长因子受体（epidermal growth factor receptor，EGFR）和Ras同源基因家族成员A（Ras homolog gene family member A，RhoA）等可能是环烯醚萜苷抗RA的核心靶点；GO和KEGG富集分析结果显示磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）/蛋白激酶B（protein kinase B，Akt）通路与环烯醚萜苷抗RA密切相关，环烯醚萜苷还参与调控中性粒细胞活化和脱颗粒等多种生物过程，进而调控机体免疫反应发挥抗RA的作用。分子对接确定了SRC为其中最核心的靶点，龙胆苦苷可能是环烯醚萜苷中的关键成分；分子动力学模拟显示龙胆苦苷与SRC靶点结合紧密，表明其具有通过靶向作用SRC对RA起治疗作用的可能性。体外实验结果证实龙胆苦苷可显著抑制HFLS-RA增殖（P＜0.01、0.001），降低上清液中炎症因子水平（P＜0.001），抑制PI3K/Akt和核因子-κB（nuclear factor-κB，NF-κB）通路的激活（P＜0.05、0.01、0.001）。结论 环烯醚萜苷是秦艽发挥药理作用的主要成分，其中龙胆苦苷为环烯醚萜苷抗RA的关键成分，可以通过抑制HFLS-RA增殖，降低炎症反应，调控PI3K/Akt和NF-κB信号通路进而发挥抗RA作用。

Objective To explore the main components and mechanism of iridoid glycosides in Gentiana straminea against rheumatoid arthritis (RA) through network pharmacology, molecular docking and experimental verification. Methods Through literature review and TCMSP data collection, iridoid glycosides were screened using SwissADME database, and drug targets were predicted using PharmMapper database; RA related targets was collected through databases such as OMIM, GeneCards and TTD, potential anti-RA targets of iridoid glycosides were screened through Venn analysis, protein-protein interaction (PPI) network was constructed to screen the core anti-RA targets of iridoid glycosides, and mechanism of iridoid glycosides anti-RA was predicted through gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. The key components of iridoid glycosides were screened through molecular docking and molecular dynamics simulations using core targets as receptors. In vitro experiments were conducted on human fibroblast-like synoviocytes in rheumatoid arthritis (HFLS-RA) to validate the results. Results A total of 18 iridoid glycosides with drug-like properties, 300 drug-related targets and 1 641 RA related targets were collected and obtained; Venn analysis screened 103 potential targets of iridoid glycosides for anti-RA; PPI network analysis revealed that proto-oncogene tyrosine-protein kinase Src (SRC), mitogen-activated protein kinase 1 (MAPK1), non-receptor tyrosine kinase lymphocyte cell-specific protein-tyrosine kinase (LCK), epidermal growth factor receptor (EGFR) and Ras homologous gene family member A (RhoA) may be the core targets of iridoid glycosides against RA. GO and KEGG enrichment analysis results showed that phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway was closely related to the anti-RA effect of iridoid glycosides, which also participated in regulating various biological processes such as neutrophil activation and degranulation, thereby regulating the body’s immune response to exert anti-RA effects. Molecular docking identified SRC was the most central target, and gentiopicroside may be a key component of iridoid glycosides; Molecular dynamics simulations showed that gentiopicroside binded tightly to the SRC target, indicating its potential therapeutic effect on RA through targeted action on SRC. The in vitro experimental results confirmed that gentiopicroside could significantly inhibit the proliferation of HFLS-RA (P < 0.01, 0.001), reduce the levels of inflammatory factors in supernatant (P < 0.001), and inhibit the activation of PI3K/Akt and nuclear factor-κB (NF-κB) pathways (P < 0.05, 0.01, 0.001). Conclusion Iridoid glycosides are the main components that exert pharmacological effects in G. straminea, among which gentiopicroside is the key component of iridoid glycosides anti-RA. It can exert anti-RA effects by inhibiting HFLS-RA proliferation, reducing inflammatory reactions and regulating PI3K/Akt and NF-κB signaling pathways.

R285.5

国家自然科学基金资助项目（81973567）；云南省万人计划基金资助项目（YNWR/MY/2020/083）；昆明市名中医专家传承工作室基金资助项目（MYGZS/2022/315）