目的 基于网络药理学分子对接及体外实验探究黄芪治疗多囊卵巢综合征（PCOS）的作用机制。方法 基于TCMSP、BATMAN、ETCM、TCMID等数据库，依据口服生物利用度（OB）和类药性（DL）标准筛选黄芪的有效成分并预测其作用靶点。通过TTD、OMIM、GeneCards数据库获取PCOS相关靶点，确定交集靶点。构建“药物-活性成分-靶点-疾病”网络模型，利用Cytoscape软件进行拓扑学分析，筛选核心成分与靶点。借助STRING数据库构建蛋白质相互作用（PPI）网络，筛选核心靶点。利用Metascape数据库进行基因本体（GO）与京都基因与基因组百科全书（KEGG）富集分析。采用分子对接技术验证核心成分与关键靶点的结合能力。以肿瘤坏死因子-α（TNF-α）构建KGN细胞PCOS模型，设对照组、模型组及不同剂量常春藤皂苷元处理组，检测细胞活力、凋亡率、脱氢表雄酮（DHEA）、睾酮、白细胞介素-1β（IL-1β）、IL-18水平及TP53、JUN基因表示水平。结果 筛选出黄芪的17种有效成分，预测其209个作用靶点，与PCOS的2 476个靶点比对后获得133个交集靶点。网络分析显示白桦脂酸、常春藤皂苷元、7-O-甲基异木犀草素等为核心成分。PPI网络分析筛选出肿瘤蛋白53（TP53）、JUN、雌激素受体1（ESR1）等核心靶点。GO富集分析显示交集靶点涉及多种生物过程、细胞组分和分子功能；KEGG富集分析揭示了磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（Akt）、丝裂原活化蛋白激酶（MAPK）等关键通路。分子对接结果显示，白桦脂酸、常春藤皂苷元、7-O-甲基异木犀草素与核心靶点TP53、JUN、ESR1均有较好结合能力，其中常春藤皂苷元与ESR1结合能最低。细胞实验表明，高剂量常春藤皂苷元可显著提升模型KGN细胞活力（P＜0.001）、降低细胞凋亡率（P＜0.05、0.001），并下调DHEA、睾酮、IL-1β、IL-18水平和TP53、JUN基因表达（P＜0.05）。结论 黄芪通过常春藤皂苷元、白桦脂酸等有效成分，结合TP53、ESR1等关键靶点，调控PI3K/Akt、MAPK等信号通路，从而改善PCOS患者的卵巢功能。

Objective To systematically explore Astragali Radix in treatment of polycystic ovary syndrome (PCOS) based on network pharmacology, molecular docking andin vitro experiments. Methods Based on the TCMSP, BATMAN, ETCM, and TCMID databases, the effective components of Astragali Radix were screened and their potential targets were predicted according to the standards of oral bioavailability (OB) and drug-likeness (DL). The targets related to PCOS were obtained through the TTD, OMIM, and GeneCards databases, and the intersecting targets were determined. A “drug-active component-target-disease” network model was constructed, and the core components and targets were screened through topological analysis using Cytoscape software. The protein interaction (PPI) network was constructed using the STRING database to screen the core targets. GO and KEGG enrichment analyses were performed using the Metascape database. The binding ability of core components to key targets was verified by molecular docking technology. A PCOS model of KGN cells was constructed using TNF-α, and groups were set up including a control group, a model group, and different doses of hederagenin treatment groups. The cell viability, apoptosis rate, and levels of DHEA, testosterone, IL-1β, and IL-18, and expression levels of TP53 and JUN genes were detected. Results A total of 17 effective components of Astragali Radix were screened out, with 209 predicted targets. After comparison with the 2 476 targets related to PCOS, 133 intersecting targets were obtained. Network analysis showed that betulinic acid, hederagenin, and 7-O-methyl luteolin were core components. PPI network analysis screened out TP53, JUN, and ESR1 as core targets. GO enrichment analysis showed that the intersecting targets were involved in various biological processes, cellular components, and molecular functions. KEGG enrichment analysis revealed key pathways such as PI3K/Akt and MAPK. Molecular docking results showed that betulinic acid, hederagenin, and 7-O-methyl luteolin had good binding ability to the core targets TP53, JUN, and ESR1, with the lowest binding energy between hederagenin and ESR1. In vitro experiments showed that high-dose hederagenin significantly increased the viability of the model KGN cells (P < 0.001), decreased the cell apoptosis rate (P < 0.05, 0.001), and downregulated the levels of DHEA, testosterone, IL-1β, IL-18, and the gene expression of TP53 and JUN (P < 0.05). Conclusion Astragali Radix improves the ovarian function of patients with PCOS by combining its effective components such as hederagenin and betulinic acid with key targets like ESR1 and TP53, and regulating the PI3K/Akt and MAPK signaling pathways.

R285.5;R914

深圳市罗湖区软科学研究计划项目（LX202402018）