运用网络药理学、分子对接与分子动力学模拟探讨逍遥丸干预乳腺癌前病变（PBC）的潜在作用机制。

检索逍遥丸活性成分及作用靶点，获取PBC相关靶点并筛选交集靶点，构建PPI网络以明确核心靶点，进一步开展GO和KEGG富集分析；选取关键活性成分与核心靶点进行分子对接，并采用GROMACS软件进行分子动力学模拟验证。

共获得逍遥丸活性成分99种、药物靶点1093个、PBC相关靶点893个及交集靶点265个。GO功能富集分析得到BP条目2524个、CC条目53个、MF条目175个；KEGG富集分析筛选出187条显著通路，主要涉及AMPK信号通路、Estrogen信号通路及肿瘤中心代谢通路等。分子对接结果显示，AKT1、PIK3CA、ESR1、CCND1等核心靶点与β-谷甾醇、芍药苷、槲皮素等关键活性成分具有较强亲和力，分子动力学模拟进一步证实其结合稳定性。

逍遥丸可能通过多成分、多靶点协同调控AKT1等核心靶点，影响AMPK/mTOR、Estrogen及肿瘤代谢相关通路，从而调节癌前细胞异常能量代谢、激素相关信号、细胞周期及免疫微环境，发挥干预PBC进展的潜在作用。本研究从“网络药理学-分子对接-分子动力学模拟”多层次验证角度，阐释了逍遥丸防治乳腺癌前病变的潜在分子机制，为经典方剂用于乳腺癌早期防治提供了新的研究依据。

To investigate the potential mechanism of Xiaoyao Pill in intervening breast precancerous lesions using network pharmacology, molecular docking, and molecular dynamics simulations.

Traditional Chinese medicine databases and platforms were used to retrieve the active components and potential targets of Xiaoyao Pill. Overlapping targets between Xiaoyao Pill and breast precancerous lesions were identified. A protein-protein interaction network was constructed, and core targets were screened from the overlapping targets. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were then performed. Key active components and core targets were subjected to molecular docking, and molecular dynamics simulations were conducted using GROMACS software to validate the network pharmacology results.

A total of 99 active components of Xiaoyao Pill, 1093 drug targets, 893 disease-related targets, and 265 overlapping targets were obtained. Gene Ontology enrichment analysis identified 2524 biological process terms, 53 cellular component terms, and 175 molecular function terms. Kyoto Encyclopedia of Genes and Genomes enrichment analysis identified 187 significantly enriched signaling pathways, primarily involving the AMPK signaling pathway, estrogen signaling pathway, and central carbon metabolism in cancer. Molecular docking showed that key targets, including AKT1, PIK3CA, ESR1, and CCND1, exhibited strong binding affinities with several core active components of Xiaoyao Pill, such as β-sitosterol, paeoniflorin, and quercetin. Molecular dynamics simulations further confirmed stable binding between the active components of Xiaoyao Pill and these targets.

Xiaoyao Pill may act on multiple signaling pathways, including the AMPK/mTOR and estrogen signaling pathways, by regulating core targets such as AKT1. It may thereby reduce abnormal energy metabolism in precancerous cells, modulate hormone-related signaling, regulate the cell cycle and immune microenvironment, and ultimately exert an interventional effect on breast precancerous lesions. This study provides new research evidence supporting the potential clinical application of this classical Chinese formula in the prevention and treatment of breast cancer.