目的 通过以网络药理学为基础的策略研究防风治疗类风湿关节炎（rheumatoid arthritis，RA）的分子生物学机制。方法 采用网络药理学方法收集防风活性成分和治疗RA的潜在靶点，并评估活性成分的药理和毒理学等相关参数；构建蛋白质相互作用网络筛选核心靶点，并通过生物信息学方法进一步验证核心靶点和疾病的关联；对核心成分和相应靶点进行分子对接。体外通过CCK-8实验、细胞迁移和侵袭、细胞凋亡、qRT-PCR和Western blotting分析，阐明别欧前胡素对MH7A细胞磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）/蛋白激酶B（protein kinase B，Akt）通路的调控作用。结果 从防风中共鉴定出18种活性成分和66个与筛选出的RA疾病靶基因相交的潜在靶基因，最终获得了汉黄芩素、β-谷甾醇、5-O-甲基维斯阿米醇和别欧前胡素等核心成分。防风治疗RA的潜在机制可能是通过调控PI3K/Akt、白细胞介素-17（interleukin-17，IL-17）、凋亡等信号通路和多种生物过程来实现，以发挥抗炎和免疫调节作用。分子对接证实了所有的核心成分和关键靶点均具有很好的对接活性。别欧前胡素抑制MH7A细胞的活力、迁移和侵袭（P＜0.05、0.01），诱导细胞凋亡（P＜0.01），并显著下调IL-1β、IL-6、IL-8、基质金属蛋白酶-1（matrix metalloproteinase-1，MMP-1）和MMP-3的基因表达（P＜0.01）。分子分析表明别欧前胡素通过抑制PI3K/Akt通路发挥对MH7A的调控作用。结论 成功预测了防风治疗RA的有效成分和潜在靶点，为进一步探究其分子机制提供了新的理论基础。揭示了别欧前胡素通过PI3K/Akt通路抑制RA成纤维样滑膜细胞的活力、迁移、侵袭及细胞因子和MMPs的表达，并诱导细胞凋亡。

Objective To investigate the molecular biological mechanism of Saposhnikovia divaricata in treating rheumatoid arthritis (RA) through a pharmacology-based strategy. Methods The bioactive phytochemicals of S. divaricata and potential targets for the treatment of RA were screened by network pharmacology, and phytochemicals-related parameters such as pharmacology and toxicology were evaluated. The protein interaction network was established to screen the core targets, and the correlation between the core targets and RA was further validated by bioinformatics strategy. Finally, molecular docking of core components and corresponding targets was performed. CCK-8 assay, cell migration and invasion, cell apoptosis, qRT-PCR, and Western blotting analysis were performed to clarify the regulation of prangenidin on phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathway in MH7A cells. Results A total of 18 bioactive phytochemicals and 66 potential target genes intersecting with the screened RA disease target genes were identified from S. divaricata. Finally, core ingredients such as wogonin, β-sitosterol, 5-O-methylvisamminol and prangenidin were obtained. The underlying mechanism of S. divaricata in treating RA might be achieved by regulating pathways such as PI3K/Akt, interleukin-17 (IL-17), apoptosis and multiple biological processes to exert anti-inflammatory and immunomodulatory effects. Molecular docking confirmed that all core ingredients and key targets had great docking activity. Prangenidin inhibited viability, migration and invasion (P < 0.05, 0.01), induced apoptosis in MH7A cells (P < 0.01), and significantly down-regulated IL-1β, IL-6, IL-8, matrix metalloproteinase-1 (MMP-1) and MMP-3 gene expressions (P < 0.01). Molecular analysis showed that prangenidin exerts its regulatory effect on MH7A cells by inhibiting PI3K/Akt pathway. Conclusion This study successfully predict the effective components and potential targets of S. divaricata in the treatment of RA, which provide a new theoretical basis for further exploring its molecular mechanism. It is revealed that prangenidin inhibits the activity, migration, invasion and expressions of cytokines and MMPs of RA fibroblast-like synovial cells through PI3K/Akt pathway, and induces cell apoptosis.

R285.5

国家自然科学基金资助项目（8217150266）；国家自然科学基金资助项目（81729003）；广州市卫生健康委员会科技项目（20211A011114）；广州市番禺区科技计划重大项目（2022-Z04-114）