基于数据挖掘、网络药理学、分子模拟和实验验证探究治疗糖尿病足溃疡外用中药复方专利的用药规律及作用机制  

Exploration of the Medication Rules and Mechanism of Patents for External Chinese Medicine Compound Formulas Against Diabetic Foot Ulcers Based on Data Mining,Network Pharmacology,Molecular Simulation and Experimental Validation

作　　者：周振华 周莉 张彬 李坤平[1] 李博 ZHOU Zhenhua;ZHOU Li;ZHANG Bin;LI Kunping;LI Bo(Institute of Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006 Guangdong,China)

机构地区：[1]广东药科大学中医药研究所,广东广州510006

出　　处：《中药新药与临床药理》2024年第4期513-523,共11页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：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)。

摘　　要：目的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-αand PPARγwere identified through PPI network

关 键 词：糖尿病足溃疡 外用中药复方专利 数据挖掘 网络药理学 分子对接 动力学模拟 实验验证 清热解毒 活血化瘀 

分 类 号：R285.5[医药卫生—中药学]