基于用药网络、靶点网络和分子对接技术的中药治疗气阴两虚兼血瘀型糖尿病肾病临床用药机制探究

Clinical Medication Mechanism of Traditional Chinese Medicine in Treatment of Diabetic Nephropathy with Qi and Yin Deficiency and Blood Stasis Based on Medication Network,Target Network,and Molecular Docking Technology

目的:通过整合用药网络及靶点网络分析中药治疗糖尿病肾病气阴两虚兼血瘀证的组方规律及其具体药理机制,为临床治疗提供科学依据并为新药研发提供参考。方法:以“糖尿病肾病”“气阴两虚”“血瘀”为主题词,在中国知网、维普资讯中文期刊服务平台、万方医学数据库的相关文献中提取符合纳入、排除标准的方药并建立方药数据库,运用古今医案云平台进行数据统计并进行用药网络相关分析,筛选出核心药对及核心处方;通过中药系统药理学数据库与分析平台(TCMSP)和GeneCards、OMIM和TTD数据库筛选出核心处方药物的有效化学成分及其相关靶点、糖尿病肾病的靶点,并取两者的交集靶点。利用Cytoscape 3.9.1软件及相关插件构建药物-有效活性成分-交集靶点-疾病网络及交集靶点蛋白质-蛋白质相互作用(PPI)网络,并进行聚类分析、基因本体(GO)功能富集分析、京都基因与基因组百科全书(KEGG)通路富集分析。结果:最终纳入229篇文献,筛选出172首方剂,涉及379味中药。治疗糖尿病肾病气阴两虚兼血瘀证方药中的高频药物药性多平、寒、微寒、温,药味多辛、甘、苦。常用配伍是丹参与黄芪、泽泻与茯苓、当归与黄芪。用药网络分析得到气阴两虚兼血瘀型糖尿病肾病的核心处方为六味地黄汤合当归补血汤加丹参、金樱子、芡实。核心处方的关键成分为槲皮素、木犀草素、山柰酚、泽泻醇B、芒柄花素等,核心靶点为细胞骨架肌动蛋白(ACTB)、Janus激酶2(JAK2)、细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子1(VCAM1)、白细胞介素-4(IL-4)等关键蛋白靶点,可能参与晚期糖基化终产物(AGEs)-晚期糖基化终产物受体(RAGE)信号通路、JAK2-信号传导及转录激活蛋白(STAT)信号通路等。分子对接结果表明,网络药理学所预测的核心靶点中的JAK2、ICAM-1、VCAM1、IL-4与关键活性成分稳定结合。结论:中医药治疗气阴两虚兼血瘀型糖尿病肾病的核心处方可通过多个活性成分,作用于多靶点、多通路发挥疗效,可为临床治疗糖尿病肾病药物的应用及研发提供参考。

Objective:To analyze the prescription rules and specific pharmacological mechanisms of traditional Chinese medicine(TCM)in the treatment of diabetic nephropathy(DN)with Qi and Yin deficiency and blood stasis by integrating the medication network and target network,so as to provide a scientific basis for clinical treatment and reference for new drug research and development.Methods:"Diabetic nephropathy""Qi and Yin deficiency",and"blood stasis"were used as the theme words to extract prescriptions that met the criteria from the relevant literature in the databases of China National Knowledge Infrastructure(CNKI),China Science and Technology Journal Database,and Wanfang Medicine,and a prescription database was established.The ancient and modern medical case cloud platform was utilized to conduct data statistics and correlation analyses of the medication network to screen out core pairs and core prescriptions.The active chemical components and related targets of the core prescriptions and the targets of DN were screened by TCMSP,Genecards,OMIM,and TTD databases,and the intersecting targets were taken.Cytoscape 3.9.1 software and related plug-ins were used to construct the drug-active component-intersecting target-disease network and protein-protein interaction(PPI)network of intersecting targets.Cluster analysis,gene ontology(GO)function enrichment analysis,and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were performed.Results:Finally,229 pieces of literature were included,and 172 prescriptions involving 379 TCM.Among the high-frequency drugs in the treatment of DN with Qi and Yin deficiency and blood stasis,the drugs were mostly plain,cold,slightly cold,and warm,and the tastes were mostly pungent,sweet,and bitter.The commonly used combinations were Salviae Miltiorrhiza Radix and Astragali Radix,Alismatis Rhizoma and Poria,and Angelicae Sinensis Radix and Astragali Radix.The core prescription in the treatment of DN with Qi and Yin deficiency and blood stasis was obtained from the analysis of the medication network as follows:Liuwei Dihuang Decoction combined with Danggui Buxue Decoction+Radix Salviae Miltiorrhiza+Rosae Laevigatae Fructus+Euryales Semen.The key components of the core prescription were quercetin,kaempferol,luteolin,alisol B,and formononetin,and the core targets were cytoskeletal actin(ACTB),Janus kinase(JAK2),intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM1),interleukin 4(IL-4),and other key protein targets,which may be involved in AGEs-RAGE signaling pathway,JAK-STAT signaling pathway,and so on.The molecular docking results showed that JAK2,ICAM-1,VCAM1,and IL-4 among the core targets predicted by network pharmacology were stably bound to the key active components.Conclusion:The core prescription of TCM for the treatment of DN with Qi and Yin deficiency and blood stasis can exert therapeutic effects through multiple active components,acting on multiple targets and pathways,which can provide a theoretical basis for the application and development of drugs for the clinical treatment of DN.