基于网络药理学和分子对接技术探讨补阳还五汤加减防治脑小血管病的作用机制

Exploring Mechanism of Buyang Huanwu Decoction(补阳还五汤) in Prevention and Treatment of Small Cerebral Vascular Disease Based on Network Pharmacology and Molecular Docking Technology

目的 基于网络药理学和分子对接技术,探讨补阳还五汤加减有效成分治疗脑小血管病(CSVD)的潜在机制。方法 从中药分子机制的生物信息学分析工具(BATMAN-TCM)和中药系统药理学数据库与分析平台(TCNSP)筛选出补阳还五汤加减的活性成分和靶点,通过在线人类孟德尔遗传(OMIM)和GeneCards数据库收集脑小血管病的靶点,将两者靶点去重后利用Venny 2.1.0制作药物和疾病的靶点韦恩图,导出其交集靶点,使用STRING数据库构建蛋白质与蛋白质相互作用的PPI网络作用图,并通过Cytoscape 3.8.2软件中的插件CytoNCA筛选关键靶点,并将交集靶点导入David数据库,利用微生信程序进行基因本体(GO)和京都和基因组百科全书(KEGG)富集分析,并通过Cytoscape3.8.2构建补阳还五汤加减药物-有效成分-靶点和补阳还五汤加减药物-成分-靶点-通路网络,最后用AutoDock vinna软件进行分子对接验证。结果 筛选得到补阳还五汤加减活性成分有80个,对应潜在241个靶点,网络图显示槲皮素、β-谷甾醇、山柰酚、豆甾醇、黄芩素为补阳还五汤加减治疗CSVD的关键成分,丝氨酸/苏氨酸蛋白应激酶1(AKT1)、白蛋白(ALB)、肿瘤坏死因子(TNF)、核内磷酸化蛋白(TP53)、血管内皮生长因子A(VEGFA)为治疗CSVD的关键靶点,GO富集分析图可能通过对基因表达的负调控、老茧对缺氧的反应,细胞组主要涉及突触、RNA聚合酶Ⅱ转录因子复合物;分子功能包括递质门控离子通道活性参与突触后膜电位调控、类固醇激素受体活性等。KEGG富集分析显示主要与非小细胞肺癌、癌症中的蛋白多糖、IL-17等信号通路有关。分子对接结果表明补阳还五汤加减中的核心作用靶点和主要活性成分具备稳定性的结合活性。结论 研究初步揭示了补阳还五汤加减可通过多靶点、多成分、多通路的机制来治疗CSVD,为临床上运用五阳还五汤加减治疗脑小血管病提供了可靠的使用依据。

Objective Based on network pharmacology and molecular docking technology,the potential mechanism of adding and reducing active ingredients of Buyang Huanwu Decoction(补阳还五汤)in the treatment of cerebral small vessel disease(CSVD)was discussed.Methods The active ingredients and targets of Buyang Huanwu Decoction were screened from the bioinformatics analysis tool of TCM molecular mechanism(BATMAN-TCM)and the TCM System pharmacology database and analysis platform(TCNSP),and the targets of cerebrovascular disease were collected through the online human Mendelian genetics(OMIM)and GeneCards databases.Venny 2.1.0 was used to make drug and disease target,and the intersection target was derived.The PPI network diagram of protein-protein interaction was constructed using STRING database,and key targets were screened by CytoNCA,a plug-in in Cytoscape 3.8.2 software.The intersection targets were introduced into the David database,and gene ontology(GO)analysis and KEGG enrichment analysis were carried out using the Weishengxin program.Moreover,the drug active ingredient target network and drug active ingredient target pathway network of Buyang Huanwu Decoction were constructed using Cytoscape 3.8.2.Finally,the molecular docking was verified by AutoDock vinna software.Result There were 80 active ingredients in Buyang Huanwu Decoction,corresponding to 241 potential targets,and the network diagram showed that quercetin,β-sitosterol,kaempferol,stigmasterol and baicalein were the key ingredients in Buyang Huanwu Decoction for CSVD treatment.AKT1(serine/threonine protein kinase 1),ALB(albumin),TNF(tumor necrosis factor),TP53(nucleophosphorylated protein),VEGFA(vascular endothelial growth factor A)were the key targets for the treatment of CSVD.GO enrichment analysis may involve negative regulation of gene expression and the response of calluses to hypoxia.The cell group mainly involved synapses and RNA polymerase II transcription factor complexes.Molecular functions included neurotransmitter gated ion channel activity involved in postsynaptic membrane potential regulation,steroid hormone receptor activity,etc.KEGG enrichment analysis showed that it is mainly related to signaling pathways such as protein polysaccharides and IL-17 in non-small cell lung cancer and cancer.The molecular docking results indicated that the core target and main active ingredients in Buyang Huanwu Decoction have stable binding activity.Conclusion This study preliminarily revealed that Buyang Huanwu Decoction can treat CSVD through the mechanism of multi-target,multi-component and multi-pathway,providing a reliable basis for the clinical use of Wuyang Huanwu Decoction in the treatment of cerebral small vascular disease.