基于UHPLC-QE-MS、网络药理学及实验验证探讨肠安菌泰治疗肝郁脾虚型腹泻型肠易激综合征的作用机制  

Study on the Mechanism of Chang’an Juntai in the Treatment of Liver Depression and Spleen Deficiency Syndrome Type of IBS-D Based on UHPLC-QE-MS,Network Pharmacology and In Vivo Experiments

作　　者：张佳河 祝旺 杨希玲 侯秋科[2] 张文杰 刘凤斌[2,4] ZHANG Jiahe;ZHU Wang;YANG Xiling;HOU Qiuke;ZHANG Wenjie;LIU Fengbin(The First Clinical Medical School of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Department of Spleen and Stomach Disease,The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Zhuhai Hospital of Guangdong Provincial Hospital of Chinese Medicine,Zhuhai 519015 Guangdong,China;Baiyun Hospital,The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510470 Guangdong,China)

机构地区：[1]广州中医药大学第一临床医学院,广东广州510405 [2]广州中医药大学第一附属医院脾胃病科,广东广州510405 [3]广东省中医院珠海医院,广东珠海519015 [4]广州中医药大学第一附属医院白云医院,广东广州510470

出　　处：《中药新药与临床药理》2025年第4期564-575,共12页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81804047,82174303);刘凤斌广东省名中医传承工作室[粤中医办函(2020)1号]。

摘　　要：目的基于超高效液相色谱-四级杆-静电场轨道阱串联质谱(UHPLC-QE-MS)、网络药理学及体内实验验证探讨肠安菌泰治疗肝郁脾虚型腹泻型肠易激综合征(IBS-D)的潜在作用机制。方法(1)采用UHPLC-QE-MS法分析肠安菌泰的化学成分;利用TCMSP数据库检索肠安菌泰9味中药的全部化学成分,并与UHPLC-QEMS鉴定出的化学成分相比对,取交集成分。筛选出肠安菌泰的活性成分并在Swiss Target Prediction数据库中进行作用靶点预测。在GeneCards、OMIM、DisGeNET数据库中检索IBS-D疾病相关基因。对肠安菌泰活性成分作用靶点与IBS-D疾病靶点取交集,筛选出肠安菌泰治疗IBS-D的潜在作用靶点。通过String数据库进行潜在作用靶点的蛋白互作(PPI)网络构建及核心靶点筛选;使用Metascape数据库对核心靶点进行GO功能及KEGG通路富集分析;利用Cytoscape软件构建“活性成分-核心靶点-通路”网络并进行拓扑分析,筛选出关键活性成分、关键靶点和关键通路;选取前3位关键活性成分与前5位关键靶点进行分子对接验证。(2)采用“母婴分离+番泻叶灌胃+醋酸灌肠+束缚应激”多因素造模方法复制肝郁脾虚型IBS-D大鼠模型。将大鼠随机分为空白组、模型组、匹维溴铵组(15.8 mg·kg^(-1))、肠安菌泰组(1.7 g·kg^(-1)),每组6只。灌胃给药,每天1次,共14 d。采用粪便含水率、腹壁撤退反射、糖水偏嗜率等指标对IBS-D动物模型进行评价;采用HE染色法观察结肠组织病理变化;RT-qPCR法检测结肠组织中PI3K、AKT mRNA表达水平;Western Blot法检测结肠组织中p-PI3K、p-AKT蛋白表达水平。结果(1)共筛选出肠安菌泰活性成分11个,获得211个潜在作用靶点和51个核心靶点,核心靶点主要与PI3K/AKT、EGFR等信号通路相关,筛选出美迪紫檀素、小檗碱、黄藤素等关键活性成分以及PIK3CB、PIK3CD、RAF1、PIK3R1、PIK3CA等关键靶点,分子对接结果显示关键活性成分与关�Objective To explore the potential mechanism of Chang’an Juntai(CAJT)in the treatment of liver stagnation and spleen deficiency syndrome type of diarrhea-predominant irritable bowel syndrome(IBS-D)based on ultra-high performance liquid chromatography Q Exactive-mass spectrometry(UHPLC-QE-MS),network pharmacology and in vivo experimental verification.Methods(1)UHPLC-QE-MS was used to analyze the chemical constituents of CAJT.The TCMSP database was used to search all the chemical constituents of 9 Chinese medicinals of CAJT,and compared with the chemical constituents identified by UHPLC-QE-MS,and the intersection components were taken.The active ingredients of CAJT were screened and the targets were predicted in the Swiss Target Prediction database.IBS-D disease-related genes were searched in GeneCards,OMIM,and DisGeNET databases.The potential targets of CAJT in the treatment of IBS-D were screened by intersection of the targets of active components of CAJT and the targets of IBS-D.The protein-protein interaction(PPI)network construction and core target screening of potential targets were carried out through the String database.Metascape database was used to analyze the GO function and KEGG pathway enrichment of core targets.Cytoscape software was used to construct the“active ingredients-core targets-pathways”network and topological analysis was performed to screen out the key active ingredients,key targets and key pathways.The first three key active components and the first five key targets were selected for molecular docking verification.(2)The IBS-D rat model of liver depression and spleen deficiency type was replicated by multi-factor modeling method of“mother-infant separation+Sennae Folium gavage+acetic acid enema+restraint stress”.The rats were randomly divided into blank group,model group,Pinaverium Bromide group(15.8 mg·kg^(-1))and CAJT group(1.7 g·kg^(-1)),with 6 rats in each group.Intragastric administration was given once a day for 14 days.The BS-D animal model was evaluated by fecal moistur

关 键 词：肠安菌泰 腹泻型肠易激综合征 肝郁脾虚型 超高效液相色谱-四级杆-静电场轨道阱串联质谱 网络药理学 分子对接 PI3K/AKT信号通路 实验验证 大鼠 

分 类 号：R285.5[医药卫生—中药学]