基于网络药理学与分子对接探究四藤一仙汤治疗糖尿病神经周围病变的作用机制

Exploring the mechanism of treating diabetes peripheral neuropathy with the Siteng Yixian decoction based on network pharmacology and molecular docking

目的:运用现代网络药理学和分子对接技术对四藤一仙汤治疗糖尿病周围神经病变的潜在有效成分进行筛选,同时推测这些成分的作用靶点,以深入研究其作用机制。方法:通过中药系统药理学数据库(TCMSP)和国家中医药百科全书数据库(ETCM)筛选四藤一仙汤中涉及的五味中药信息及潜在的药理作用和靶点信息,通过在人类基因数据库(GeneCards)中搜索与糖尿病周围神经病变疾病相关的靶点信息,然后将其与四藤一仙汤中的成药靶点信息取交集,得到成药-疾病交集靶点。使用STRING平台构建这些交集靶点的蛋白质-蛋白质相互作用网络,并将蛋白质-蛋白质相互作用网络数据以TSV文件格式导入Cytoscape 3.8.2软件。在Cytoscape 3.8.2软件中,我们将进行关键靶点分析,依据连接度值进行二次建模,进行亮暗颜色分析,从而检测出最关键的核心靶点;此外利用Metascape数据库分析药物-疾病靶点互相作用网络,同时完成对基因本体论(GO)和京都基因与基因组百科全书(KEGG)的信息的富集与分析,从而筛选出四藤一仙汤治疗糖尿病周围神经病变的活性物质成分。最后用Autodock软件和Pymol软件对主要活性成分及核心靶点进行分子对接验证和可视化展示。结果:药物和疾病的潜在靶点分别为249个和1324个,它们之间的交集靶点为108个。经过筛选,我们获得了核心靶点,其中包括信号转导因子和转录激活因子(Signal Transducer and Activator of Transcription,STAT)3、丝氨酸/苏氨酸蛋白激酶1(Akt Serine/Threonine Kinase 1,AKT1)、肿瘤蛋白p53(Tumor Protein p53,TP53)等。GO功能富集分析显示,四藤一仙汤对细胞生物学过程具有多种影响,包括细胞对氮化合物的反应、对无机物的反应、细胞迁移的正向调节、细胞对脂质的反应、蛋白质磷酸化的正调节、细胞对有机环状化合物的反应、细胞群增殖的负调控、细胞分解代谢过程的调节等。同时,KEGG通路富集分析显示,四藤一仙汤可能通过多种通路发挥治疗作用,其中包括癌症的途径、晚期糖基化终末产物(Advanced Glycation End-product,AGE)-糖基化终末产物受体(Receptor for Advanced Glycation End Products,RAGE)信号通路在糖尿病并发症中的作用、脂质和动脉粥样硬化、人类巨细胞病毒感染、丝裂原活化蛋白激酶(Mitogen-Activated Protein Kinase,MAPK)信号通路、化学致癌-活性氧、糖尿病性心肌病、胰岛素抵抗、疟疾、松弛素信号通路等。分子对接结果显示AKT1与β-谷甾醇、TP53与木犀草素有较好的结合活性。结论:四藤一仙汤在治疗糖尿病周围神经病变的过程中,表现出多靶点、多基因、多通路的作用机制。除了减轻疼痛作用外,它还具有一定的减轻血管内皮损害、改善周围神经和预防癌症的效果。这些发现为进一步研究和使用这一经典验方治疗糖尿病周围神经病变提供了新的研究方向。

Objective:Using modern network pharmacology and molecular docking techniques,we conducted a screening of potential effective components of the Siteng Yixian decoction(四藤一仙汤)for the treatment of diabetic peripheral neuropathy Methods:In the Siteng Yixian decoction,five TCM medicine components were subjected to screening using the TCMSP as well as the ETCM.The aim was to identify their potential pharmacological effects and target information.Furthermore,relevant target information for DPN was obtained by searching the GeneCards database.By taking the intersection of these two sets of data,we obtained the drug-disease intersection targets.These intersection targets can be visualized using a Venn diagram to highlight the commonalities and distinctions between the traditional Chinese medicine components and DPN targets.Use the STRING platform to construct the PPI network of the intersection targets,and convert the PPI network TSV file import Cytoscape 3.8.2 software,use Cytoscape 3.8.2 software for key target analysis,carry out secondary modeling according to degree value,and carry out light and dark color analysis,so as to detect the most critical core targets.In addition,the Metascape database can also be used at the same time.The enrichment and analysis of the information of GO and KEGG can detect the active substances of the Siteng Yixian decoction in the treatment of DPN.Finally,molecular docking validation and visualization of the major active compounds and core targets were performed using Autodock software and Pymol software.Results:A total of 249 potential targets for drugs and 1324 potential targets for diseases were obtained,and 108 targets were intersected.The core targets after screening:STAT3,AKT1,TP53,etc..GO functional enrichment analysis obtained cell biological processes including:cellular response to nitrogen compound,response to inorganic substance,positive regulation of cell migration,cellular response to lipid,positive regulation of protein phosphorylation,cellular response to organic cyclic compound,negative regulation of cell population proliferation,regulation of cellular catabolic process,etc..KEGG pathway enrichment analysis obtained:pathways in cancer,AGE-RAGE signaling pathway in diabetic complications,lipid and atherosclerosis,human cytomegalovirus infection,MAPK signaling pathway,chemical carcinogenesis-reactive oxygen species,etc..Molecular docking results demonstrate that AKT1 exhibits favorable binding activity withβ-sitosterol,while TP53 shows strong affinity with piceatannol.Conclusion:Through network pharmacology analysis,the Siteng Yixian decoction can be expressed through multi-target,multi gene,multi pathway during the treatment of diabetic peripheral neuropathy,which in addition to its pain reducing effects,has a certain effect of alleviating vascular endothelial damage,improving peripheral nerves,and preventing cancer.To provide new research directions for the future use of classical experimental methods for the treatment of DPN.