目的 探讨补骨脂总黄酮治疗类风湿关节炎的药效及作用机制。方法 采用极性分步提取制备补骨脂总黄酮，并进行HPLC分析及含量测定；通过网络药理学与分子对接筛选核心靶点与通路；以胶原诱导型关节炎小鼠观察补骨脂总黄酮的体内抗炎及关节保护作用，以细胞实验考察其对类风湿关节炎滑膜成纤维（RA-FLS）细胞增殖、迁移、侵袭的影响；采用Western blotting检测磷酸化蛋白激酶B（p-Akt）、Akt、磷酸化糖原合成酶激酶-3β（p-GSK-3β）、GSK-3β、β-连环蛋白（β-catenin）的表达水平。结果 补骨脂总黄酮主要含有新补骨脂异黄酮、补骨脂二氢黄酮甲醚和异补骨脂查尔酮等8种成分；网络药理学预测得到补骨脂总黄酮和类风湿关节炎的48个共同靶点，筛选得到肿瘤坏死因子（TNF）、白细胞介素（IL）-6、Akt1等关键靶点，基因本体论（GO）分析和京都基因和基因组百科全书（KEGG）通路富集分析结果显示，补骨脂总黄酮可能作用于PI3K/Akt、丝裂原活化蛋白激酶（MAPK）、低氧诱导因子-1（HIF-1）和TNF等通路；分子对接显示主要成分与Akt1对接亲和力良好。体内实验表明补骨脂总黄酮可显著降低模型小鼠关节炎评分、足肿胀度及关节病理损伤（P＜0.05、0.01）；体外实验显示，补骨脂总黄酮显著抑制RA-FLS增殖、迁移及侵袭（P＜0.05、0.001），补骨脂总黄酮2.0 μg/mL组p-Akt/Akt、p-GSK-3β/GSK-3β、β-catenin的蛋白表达水平明显下调（P＜0.05、0.01）。结论 补骨脂总黄酮可通过抑制Akt/GSK-3β/β-catenin通路，减轻滑膜炎症与异常增殖，缓解关节损伤，发挥抗类风湿关节炎作用。

Objective To explore the efficacy and action mechanism of total flavonoids from Psoralea corylifolia in treatment of rheumatoid arthritis. Methods Total flavonoids from Psoralea corylifolia was prepared by stepwise extraction with polar solvents and analyzed by HPLC for content determination. Network pharmacology and molecular docking were used to screen core targets and pathways. The anti-inflammatory and joint-protective effects of total flavonoids from Psoralea corylifolia in vivo were observed in collagen-induced arthritis mice, and its effects on the proliferation, migration and invasion of RA-FLS cells were investigated in vitro. The expression levels of p-Akt, Akt, p-GSK-3β, GSK-3β, and β-catenin were detected by Western blotting. Results Total flavonoids from Psoralea corylifolia mainly contains 8 major constituents such as neobavaisoflavone, bavachin, and isobavachalcone. Network pharmacology predicted 48 common targets of total flavonoids from Psoralea corylifolia and rheumatoid arthritis, and key targets such as TNF, IL-6, and Akt1 were screened. GO analysis and KEGG pathway enrichment analysis showed that total flavonoids from Psoralea corylifolia may act on pathways such as PI3K/Akt, MAPK, HIF-1, and TNF. Molecular docking showed that the main constituents had good binding affinity with Akt1. In vivo experiments showed that total flavonoids from Psoralea corylifolia could significantly reduce the arthritis score, paw swelling degree and joint pathological damage in model mice (P < 0.05, 0.01). In vitro experiments showed that total flavonoids from Psoralea corylifolia significantly inhibited the proliferation, migration and invasion of RA-FLS (P < 0.05, 0.001), and significantly down-regulated the protein expression levels of p-Akt/Akt, p-GSK-3β/GSK-3β, and β-catenin (P < 0.05, 0.01). Conclusion Total flavonoids from Psoralea corylifolia can exert anti- rheumatoid arthritis effects by inhibiting the Akt/GSK-3β/β-catenin pathway, reducing synovial inflammation and abnormal proliferation, and alleviating joint damage.

国家自然科学基金项目（82374179）；中国中医科学院中药研究所人才引进项目（ZXKT22057）；中国中医科学院优秀青年科技人才培养专项（ZZ17-YQ-024）；中国中医科学院基本科研业务费新入职科研人员启动资金专项（ZZ16-XRZ-070）