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 性成分与关键靶点均有较好的结合活性。（2）与空白组比较，模型组大鼠的粪便含水率显著升高（P＜0.001），腹 壁撤退反射及糖水偏嗜率显著降低（P＜0.01）；结肠组织 PI3K、AKT mRNA 表达水平显著升高（P＜0.001）， p-PI3K、p-AKT 蛋白表达显著上调（P＜0.001）。与模型组比较，肠安菌泰组和匹维溴铵组大鼠的粪便含水率 显著降低（P＜0.001），腹壁撤退反射及糖水偏嗜率显著升高（P＜0.01，P＜0.001）；结肠组织 PI3K、AKT mRNA 表达水平显著降低（P＜0.001），p-PI3K、p-AKT 蛋白表达显著下调（P＜0.001）。结论 肠安菌泰可能通 过美迪紫檀素、小檗碱、黄藤素等关键活性成分，作用于 PIK3CB、PIK3CD、RAF1、PIK3R1、PIK3CA 等关 键靶点，调控 PI3K/AKT 等关键信号通路，发挥治疗 IBS-D 的作用。

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.

R285.5

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］。