目的 通过网络药理学和实验验证分析六味地黄丸改善卵巢功能下降的有效成分和分子机制。方法 利用中药系统药理学数据库与分析平台（TCMSP）、Swisstarget、Uniprot数据库筛选六味地黄丸的有效化学成分和靶点，从Genecards、DisGeNET数据库中获取卵巢储备功能下降和早发性卵巢功能不全的相关靶点，制成Venn图得到药物-疾病的交集靶点。使用STRING进行蛋白质-蛋白质相互作用（PPI）分析，并使用Cytoscape软件构建PPI网络和药物成分-靶标网络，使用DAVID数据库对交集基因进行基因本体（GO）注释及京都基因与基因组百科全书（KEGG）通路富集分析。进行分子对接以确定生物活性组分与靶标之间的结合活性。通过环磷酰胺制备卵巢功能下降小鼠模型，对网络药理学分析所预测的六味地黄丸干预卵巢功能下降的潜在机制进行实验验证。结果 六味地黄丸共筛选出74种活性成分、485个潜在靶点。卵巢功能下降与六味地黄丸之间存在123个共同的治疗靶点，通过对这些共同靶点的进一步分析，共富集出1 307个GO过程以及181条KEGG信号通路。分子对接结果显示六味地黄丸主要有效成分尤其是泽泻醇B与关键靶点具有良好的结合能力。动物实验结果表明六味地黄丸可显著影响卵巢功能下降模型小鼠的磷脂酰肌醇3-激酶（PI3K）、丝氨酸和苏氨酸激酶（AKT）、表皮生长因子受体（EGFR）基因和蛋白的表达。结论 六味地黄丸能够通过其多种成分作用于多个靶点，并通过多种途径对抗卵巢功能减退。PI3K/Akt信号通路和EGFR靶点在治疗过程中发挥重要作用。

Objective To analyze the active ingredients and molecular mechanisms by which Liuwei Dihuang Pills improves decreased ovarian function through network pharmacology and experimental validation. Methods The TCMSP, Swisstarget, and Uniprot databases were used to screen Liuwei Dihuang Pills’ active chemical components and targets. Relevant targets for diminished ovarian reserve and premature ovarian insufficiency (POI) were obtained from the Genecards and DisGeNET databases, and a Venn diagram was created to identify overlapping drug-disease targets. Protein-protein interaction (PPI) analysis was performed using STRING, and PPI networks and drug component-target networks were constructed using Cytoscape software. GO and KEGG enrichment analyses were conducted on the intersecting genes through the DAVID database. Molecular docking was used to determine the binding activity between bioactive components and targets. The potential mechanisms of Liuwei Dihuang Pills on decreased ovarian function predicted by network pharmacology were experimentally validated. Results A total of 74 active ingredients and 485 potential targets of Liuwei Dihuang Pills were identified. There were 123 shared therapeutic targets between decreased ovarian function and Liuwei Dihuang Pills. Further analysis of these common targets enriched 1 307 GO items and 181 KEGG pathways. Molecular docking results showed that the main compounds of Liuwei Dihuang Pills, especially alisol B, had good binding ability with key targets. Animal experiments indicated that Liuwei Dihuang Pills significantly affected the expression of PI3K, Akt, and EGFR genes and proteins. Conclusion Liuwei Dihuang Pills can act on multiple targets through various components and counteract ovarian function decline through multiple pathways. Additionally, the study highlights the potential importance of the PI3K/Akt signaling pathway and EGFR targets in the treatment process.

R285.5

国家自然科学基金资助项目（82260947）；宁夏自然科学基金资助项目（2023AAC05059）