目的 以网络药理学研究结果为基础，对小建中汤中桂枝、甘草、生姜、大枣、白芍、饴糖6种中药开展深入研究，以期为小建中汤治疗脾胃虚寒类消化性溃疡作用机制研究提供新的依据及方向。方法 基于中药系统药理学数据库与分析平台(TCMSP)、中医药综合数据库(TCMID)获取小建中汤6种中草药的相关化学成分，并在Swiss Target Prediction中搜索活性化学成分潜在靶点;通过GeneCards数据库检索消化性溃疡、十二指肠溃疡、胃溃疡靶点和脾胃虚寒型消化性溃疡主要症状靶点;筛选出疾病主要症状、疾病和药物的共同靶点;运用Cytoscape Version 3。7。2软件绘制药物-化合物-靶点网络;利用STRING数据库绘制构建蛋白质相互作用(PPI)网络分析共同靶点之间的相互作用，找出可能的关键基因;利用R软件进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析，分析治疗的作用机制;运用AutoDock及Pymol对关键靶点与相应活性成分进行分子对接验证，实验验证小建中汤对靶点蛋白、白细胞介素(IL)-6表达的影响。结果 分析得到小建中汤具有128个活性化学成分作用于75个脾胃虚寒型消化性溃疡靶点，其中TP53、血管内皮生长因子(VEGF)、IL-6、胱天蛋白酶3(CASP3)、丝氨酸/苏氨酸蛋白激酶1(AKT1)、信号转导因子和转录激活因子3(STAT3)、丝裂原活化蛋白激酶3(MAPK3)、白蛋白(ALB)、表皮生长因子受体(EGFR)与非受体酪氨酸激酶(SRC)为核心靶点;而小建中汤治疗脾胃虚寒型消化性溃疡的核心活性成分是儿茶素、羟黄酮、芍药内苷C、拟雌内酯、草果甲素等;靶点基因主要富集于脂筏、膜微结构域、膜区、细胞小凹等细胞区域，作用于跨膜受体蛋白酪氨酸激酶活性、蛋白酪氨酸激酶活性、跨膜受体蛋白激酶活性、磷酸酶结合等分子功能，重要的是具有调节平滑肌细胞增殖和氧化应激反应。生物学通路则主要集中于磷脂酰肌醇3-激酶(PI3K)-Akt信号通路、缺氧诱导因子(HIF)-1信号通路、C型凝集素受体信号通路、胃癌等通路。分子对接结果显示，筛选的主要核心成分与其对应靶蛋白具有较好的结合活性。结论 小建中汤通过网络药理学预测的作用机制多成分、多靶点、多通路治疗脾胃虚寒型消化性溃疡;实验验证小建中汤可下调靶点蛋白IL-6的表达，抑制肠上皮损伤。
Network pharmacology research and validation of the mechanism of Xiaojianzhong Tang for the treat-ment of spleen-stomach deficiency-cold type peptic ulcers
Objective To provide new evidence and direction for the mechanism of Xiaojianzhong Tang in treating spleen-stomach deficiency-cold type peptic ulcers by investigating the six traditional Chinese medicines in Xiaojianzhong Tang,including Guizhi,Licorice,Ginger,Jujube,White Peony,and Maltose based on the re-search results of network pharmacology.Methods The relevant chemical components of the six herbs in Xiao-jianzhong Tang were retrieved using TCMSP data and TCMID database.Subsequently,we employed Swiss Target Prediction to identify potential targets of the active chemical constituents.Then we further conducted searches in the Gene Cards database to retrieve targets related to peptic ulcer,duodenal ulcer,gastric ulcer,as well as targets associated with the main symptoms of spleen-stomach deficiency-cold type peptic ulcers.Finally,the common tar-gets of the disease and the drugs were screened out.The drug-compound-target network was drawn using Cy-toscape Version 3.7.2 software.We constructed a PPI network using the STRING database to analyze the interac-tions among these common targets,and identify possible key genes.GO and KEGG enrichment analyses were per-formed using R software to preliminarily investigate the therapeutic mechanisms of action.The molecular docking validation of key targets with corresponding active ingredients was conducted using AutoDock and Pymol.Further-more,experimental validation was conducted to investigate the impact of Xiaojianzhong Tang on the expression of the target protein IL-6.Results The analysis indicated that Xiaojianzhong Tang contained 128 active chemical constituents targeting 75 gastric ulcer-related targets of the spleen-stomach deficiency-cold type.Among them,TP53,VEGF,IL-6,CASP3,AKT1,STAT3,MAPK3,ALB,EGFR,and SRC were the core targets.The core active ingredients of Xiaojianzhong Tang in treating spleen-stomach deficiency-cold type peptic ulcers were Catechin,(+)-Catechin,Phenol,Paeonilactone C,Coumestrol,and Melilotocarpan A.The target genes were mainly enriched in cell regions such as lipid rafts,membrane microdomains,membrane regions,and cell small pits,acting on molecu-lar functions such as transmembrane receptor protein tyrosine kinase activity,protein tyrosine kinase activity,transmembrane receptor protein kinase activity,and phosphatase binding.Importantly,these target genes played a role in regulating smooth muscle cell proliferation and oxidative stress response.The biological pathways mainly focused on PI3K Akt signaling pathway,HIF-1 signaling pathway,C-type lectin receptor signaling pathway,and gastric cancer-related signaling pathway.The molecular docking results indicated that the selected core compo-nents exhibited favorable binding activity with their corresponding target proteins.Conclusion Xiaojianzhong Tang can treat spleen-stomach deficiency-cold type gastric ulcers through a mechanism predicted by network pharmacology involving multiple components,targets,and pathways.Xiaojianzhong Tang can downregulate the ex-pression of the target protein IL-6 and inhibit intestinal epithelial.
万方数据
维普
