中药新药与临床药理2025，Vol.36Issue(4)：564-575,12.DOI:10.19378/j.issn.1003-9783.2025.04.009
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
摘要
Abstract
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 moisture content,abdominal withdrawal reflex and sucrose preference rate.The pathological changes of colon tissue were observed by HE staining.The mRNA expression levels of PI3K and AKT in colon tissue were detected by RT-qPCR.The protein expression levels of p-PI3K and p-AKT in colon tissues were detected by Western Blot.Results(1)A total of 11 active ingredients of CAJT were screened,211 potential targets and 51 core targets were obtained.The core targets were mainly related to PI3K/AKT,EGFR and other signaling pathways.The key active ingredients such as Medicarpin,Berberine and Palmatine,as well as key targets such as PIK3CB,PIK3CD,RAF1,PIK3R1 and PIK3CA were screened out.Molecular docking results showed that the key active ingredients had good binding activity with the key targets.(2)Compared with the blank group,the fecal moisture content of the rats in the model group was significantly increased(P<0.001),and the abdominal withdrawal reflex and sugar water preference rate were significantly decreased(P<0.01).The mRNA expressions of PI3K and AKT in colon tissue was significantly increased(P<0.001),and the protein expressions of p-PI3K and p-AKT was significantly upregulated(P<0.001).Compared with the model group,the fecal moisture content of the rats in the CAJT group and the pinaverium bromide group was significantly decreased(P<0.001),and the abdominal withdrawal reflex and sucrose preference rate were significantly increased(P<0.01,P<0.001).The mRNA expressions of PI3K and AKT in colon tissue was significantly decreased(P<0.001),and the protein expressions of p-PI3K and p-AKT was significantly down regulated(P<0.001).Conclusion CAJT may act on key targets such as PIK3CB,PIK3CD,RAF1,PIK3R1 and PIK3CA through key active ingredients such as Medicarpin,Berberine,Palmatine.CAJT may be used for the treatment of IBS-D by inhibiting the activation of the PI3K/AKT signaling pathway.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Key words
Chang'an Juntai/IBS-D/UHPLC-QE-MS/liver stagnation and spleen deficiency type/UHPLC-QE-MS/network pharmacology/molecular docking/PI3K/AKT signaling pathway/experimental validation/rats分类
医药卫生引用本文复制引用
张佳河,祝旺,杨希玲,侯秋科,张文杰,刘凤斌..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[J].中药新药与临床药理,2025,36(4):564-575,12.基金项目
国家自然科学基金项目(81804047,82174303) （81804047,82174303）
刘凤斌广东省名中医传承工作室[粤中医办函(2020)1号]. （2020）
