基于国家知识产权局专利数据库，分析外用中药复方专利治疗糖尿病足溃疡（diabetic foot ulcer， DFU）的用药规律及作用机制，并通过实验验证相关结果。方法 根据纳入和排除标准检索数据库中治疗DFU 的外用中药复方专利，去重后构建DFU 外用中药复方专利集，对其进行频数、性味归经、关联规则及聚类分 析，筛选治疗DFU 的外用核心中药数据集；进一步建立疾病-核心中药-活性成分-共有靶点网络模型，通过共 有靶点建立蛋白-蛋白互作网络（protein-protein interaction networks，PPI），利用Metascape 平台进行基因本体 （gene ontology，GO）富集分析与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG） 通路分析；借助分子对接及动力学模拟评估潜在关键靶标与活性成分之间的结合能力与稳定性；并基于CCK-8 法评估关键活性成分对细胞增殖的影响，利用实时荧光定量PCR（quantitative real-time PCR，qPCR）验证活性 成分对PI3K 及Akt 信号通路的调控作用。结果 共收集专利98 项，包含中药281 味。其中，寒性中药应用次 数最多，归肝经中药应用最多，清热类中药应用最多。通过数据挖掘获得频数前3 位的核心中药为当归、黄柏 和红花，进一步筛选得出核心中药包含活性成分85 种，其对应疾病靶点889 个；与DFU 靶点取交集后得到 166 个共有靶点，通过PPI 网络拓扑分析，确定了包括肿瘤坏死因子α（TNF-α）、过氧化物酶体增殖物激活受 体γ（PPARγ）等8 个核心靶点。GO 和KEGG 通路富集分析证实，核心中药的活性成分主要作用于炎症反应及 细胞凋亡等相关靶点，通过调控磷脂酰肌醇3-激酶/蛋白激酶B（PI3K/Akt）等信号通路治疗DFU。分子对接和 动力学模拟表明关键靶点与其对应的核心成分之间具有良好的结合能力及稳定性。实验研究表明，关键活性成 分豆甾醇具有较高的生物安全性，其可明显上调PI3K 及Akt 的转录水平。结论 通过多维数据挖掘分析证实 治疗DFU 的外用复方大多具有清热解毒、活血化瘀特性，其关键活性成分豆甾醇、二氢尼洛替星主要作用于 PPARγ 及TNF-α 等DFU 靶点；豆甾醇可明显上调PI3K 及Akt 信号通路发挥治疗DFU 的效果。

The medication rules and mechanism of patents for external Chinese medicine compound formulas against diabetic foot ulcers（DFU） in National Intellectual Property Administration were analyzed and the results were validated by experiments. Methods The database of patents for external Chinese medicine compound formulas on the treatment of DFU were retrieved according to the inclusion and exclusion criteria. Patent set of external Chinese medicine compound formulas on the treatment of DFU were constructed after deduplication. The frequency， properties， flavors， meridian tropism， association rules and cluster analysis were further performed to screen the core external Chinese medicine datasets in the treatment of DFU. Then network model of disease- core Chinese medicinals-active ingredients-common targets was constructed. Protein-protein interaction （PPI） networks were built via common targets. Gene oncology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis were performed by Metascape platform. Molecular docking and dynamics simulations were employed to evaluate the binding affinity and stability between key targets and active ingredients. The effect of active ingredients on cell proliferation were analyzed by CCK-8， and qPCR was used to evaluate the effect of active ingredients on PI3K and Akt signaling pathway. Results A total of 98 patents for external Chinese medicine compound formulas on the treatment of DFU were screened， including 281 Chinese medicinals. Among them， Chinese herbal medicines in cold property are the most used drugs. Heat-clearing Chinese herbs and Chinese herbs for liver meridians are widely used. Data mining revealed that Angelica Sinensis Radix， Phellodendri Chinese Cortex， and Carthami Flos were top 3 core drugs in terms of frequency. A total of 85 active ingredients and 889 targets of the core drugs were obtained. Then， 166 shared targets were screened after intersecting with DFU targets and targets of the core drugs. Eight core targets，including TNF-α and PPARγ were identified through PPI network topology analysis. GO and KEGG pathway enrichment analysis proved that active compounds of core drugs mainly acted on inflammation and cell apoptosis via multiple -signaling pathways，such as PI3K/Akt to treat DFU. Molecular docking and dynamics simulations demonstrated that there were good binding affinities and stabilities between core targets and their corresponding active ingredients. Meanwhile， stigmasterol shown high biosafety and enhanced the transcriptional level of PI3K and Akt in experimental validation. Conclusion In summary，this study confirmed that the essential characteristic of external compound in treating DFU were clearing away heat and removing toxins， invigorating blood and dissolving stasis. The key active ingredients， stigmasterol and dihydroniloticin， mainly targeted PPARγ and TNF - α. Stigmasterol could obviously upregulate PI3K and Akt signaling pathway to display the therapeutic effect on DFU.

R285.5

广东省中医药局科研项目（20241171，20244044）；广东省医学科研基金项目（A2022168）。