广西医学

目的基于网络药理学和分子对接技术探究黄芪-地黄药对治疗糖尿病的作用机制。方法通过中药系统药理学数据库分析平台及中医药整合药理学研究平台分别筛选得到黄芪、地黄的活性化学成分，再利用SwissTargetPrediction数据库获取活性化学成分的作用靶点。通过GeneCard数据库、DisGeNET数据库检索糖尿病相关靶点。将上述靶点取交集，利用STRING平台和 Cytoscape 软件构建交集靶点的蛋白-蛋白相互作用（PPI）网络以筛选出核心靶点。运用Cytoscape软件构建药物-成分-靶点-疾病网络以筛选核心活性化学成分。采用Metascape软件对交集靶点进行功能富集分析和通路富集分析。采用AutoDock Tools 1.5.7软件对核心活性化学成分与核心靶点进行分子对接分析。结果共筛选到12个地黄活性化学成分和13个黄芪活性化学成分，活性化学成分的作用靶点582个，糖尿病相关靶点1 646个，交集靶点176个。黄芪-地黄药对治疗糖尿病的核心靶点为丝氨酸/苏氨酸蛋白激酶（AKT）1、肿瘤坏死因子（TNF）、GAPDH、表皮生长因子受体（EGFR）、原癌基因酪氨酸蛋白激酶（SRC），核心活性化学成分为3,9-di-O-美迪紫檀素、华良姜素、京尼平苷、异鼠李素、山柰酚、槲皮素。交集靶点涉及对激素的反应、对无机物的反应、细胞对含氮化合物的反应等生物过程，涉及膜筏、膜微区、细胞质的核周区域等细胞组分，涉及以醇基为受体的磷酸转移酶活性、蛋白激酶活性、激酶活性等分子功能，以及涉及磷脂酰肌醇3-激酶(PI3K)/AKT信号通路、胰岛素信号通路、胰岛素抵抗、糖尿病并发症中的晚期糖基化终末产物(AGE)-糖基化终末产物受体(RAGE)信号通路、TNF信号通路、低氧诱导因子1（HIF-1）信号通路等信号通路。分子对接结果显示，所有核心活性化学成分与核心靶点均结合良好。结论黄芪-地黄药对的3,9-di-O-美迪紫檀素、华良姜素、京尼平苷、异鼠李素、山柰酚、槲皮素等活性化学成分可能通过作用于AKT1、TNF、GAPDH、EGFR和SRC等靶点，调控PI3K/AKT信号通路、胰岛素信号通路、胰岛素抵抗、糖尿病并发症中的AGE-RAGE信号通路、HIF-1信号通路、TNF信号通路等，从而发挥治疗糖尿病的作用。

ObjectiveTo explore the mechanism of Hedysarum multijugum-Rehmanniae radix praeparata drug pair for the treatment of diabetes mellitus based on network pharmacology and molecular docking technique. MethodsThe active chemical components of Hedysarum multijugum and Rehmanniae radix praeparata were screened by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine, respectively, and then the effect targets of active chemical components were obtained through the SwissTargetPrediction database. Targets related to diabetes mellitus were retrieved from the GeneCards and DisGeNET databases. The intersection of aforementioned targets was obtained, and the protein-protein interaction (PPI) network of intersection targets was established by using the STRING platform and Cytoscape software. The Cytoscape software was used to establish drugs-components-targets-diseases network for screening core active chemical components. The functional enrichment analysis and pathway enrichment analysis were performed on intersection targets by employing the Metascape software. The AutoDock Tools 1.5.7 software was used to perform molecular docking analysis on the core active chemical components and the core targets. ResultsA total of 12 active chemical components of Rehmanniae radix praeparata and 13 active chemical components of Hedysarum multijugum were screened; moreover, there were 582 effect targets of active chemical components, 1646 targets related to diabetes mellitus, and 176 intersection targets. The core targets of Hedysarum multijugum-Rehmanniae radix praeparata drug pair for the treatment of diabetes mellitus were serine/threonine protein kinase (AKT)1, tumor necrosis factor (TNF), GAPDH, epidermal growth factor receptor (EGFR), proto-oncogene tyrosine-protein kinase (SRC), and the core active chemical components were 3,9-di-O-methylnissolin, jaranol, geniposide, isorhamnetin, kaempferol, and quercetin. Biological processes involved in the intersection targets were response to hormone, response to inorganic compound, and cell responses to nitrogenous complexes, etc.,cellular compositions in the intersection targets were membrane raft, membrane microdomain, and perinuclear region of cytoplasm, etc.,molecular functions in the intersection targets were phosphotransferase activity with alcohol group as receptor, protein kinase activity, kinase activity, etc., and signaling pathways in the intersection targets were phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway, insulin signaling pathway, insulin resistance, signaling pathway of advanced glycation end product (AGE)-receptor for AGE (RAGE) in diabetic complications, TNF signaling pathway, hypoxia inducible factor 1 (HIF-1) signaling pathway, etc. The results of molecular docking revealed that all core active chemical components were in favorable binding with the core targets. ConclusionThe active chemical components of Hedysarum multijugum-Rehmanniae radix praeparata drug pair in terms of 3,9-di-O-methylnissolin, jaranol, geniposide, isorhamnetin, kaempferol, and quercetin, etc., may regulate PI3K/AKT signaling pathway, insulin signaling pathway, insulin resistance, AGE-RAGE signaling pathway in diabetic complications, HIF-1 signaling pathway, and TNF signaling pathway, etc.,so as to exert therapeutic effects on diabetes mellitus through acting on AKT1, TNF, GAPDH, EGFR, SRC, and other targets.