临床肝胆病杂志2025，Vol.41Issue(12)：2586-2596,11.DOI:10.12449/JCH251221
基于网络药理学探讨柴胡皂苷e联合吉非替尼治疗胆管癌的作用机制
Mechanism of action of saikosaponin e combined with gefitinib in treatment of cholangiocarcinoma:A study based on network pharmacology
摘要
Abstract
Objective To investigate the pharmacological effect and molecular mechanism of saikosaponin e combined with gefitinib in the treatment of cholangiocarcinoma based on network pharmacology and cell experiment.Methods SwissTargetPrediction was used to obtain the drug action targets of saikosaponin a,b1,c,d,e,f,g,and h,and GeneCards was used to obtain the targets of cholangiocarcinoma.The intersecting targets of these two groups of targets were imported into STRING to construct a protein-protein interaction network.WEB-based GEne SeT AnaLysis Toolkit was used to perform gene ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis and construct a target-pathway network,and Pathview R package was used to label the targets covered by the key pathways.CytoScape 3.7.2 was used to construct a drug-disease-target network,and molecular docking was performed between effective compounds and key targets.Human cholangiocarcinoma RBE cells were randomly divided into control group,saikosaponin e group,gefitinib group,and saikosaponin e+gefitinib group.MTT assay,EdU,scratch assay,and the fluorescence probe method were used to measure the proliferation and migration of RBE cells and the production of reactive oxygen species(ROS)and malondialdehyde(MDA),and Western blotting was used to measure the protein expression levels of phosphatidylinositol 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(AKT),and phosphorylated AKT(p-AKT).Logistic regression was used to calculate half-maximal inhibitory concentration(IC50);A one-way analysis of variance and repeated measures analysis of variance were used for comparison of continuous data between multiple groups,and the Tukey test was used for further comparison between two groups;the least significant difference t-test was used for comparison of simple effect.Results A total of 34 saikosaponin targets and 1 815 cholangiocarcinoma targets were obtained,resulting in 12 intersecting targets.The topological analysis showed that saikosaponins e and c had a stronger efficacy against cholangiocarcinoma,with the key targets of STAT3,IL-2,and PPP2CA,and saikosaponin e and c could respectively dock with the corresponding amino acid sites of STAT3,IL-2,and PPP2CA by forming hydrogen bonds.The GO functional enrichment analysis obtained 13 biological processes,16 cellular components,and 13 molecular functions,and the KEGG pathway enrichment analysis obtained 9 pathways,among which PI3K-AKT and EGFR tyrosine kinase inhibitor resistance were the key signaling pathways,covering multiple targets including STAT3,IL-2,and PPP2CA.Saikosaponin e and gefitinib had an IC50 of 16.89 µmol/L and 27.49 µmol/L,respectively,on human cholangiocarcinoma RBE cells at 24 hours of treatment,and compared with the control group,saikosaponin e treatment for 24 hours significantly reduced the proliferation rate(53.46%±6.42%vs 100.00%±6.00%,P<0.000 1)and migration rate(12.06%±1.76%vs 16.01%±1.89%,P<0.05)of human cholangiocarcinoma RBE cells,increased the production of ROS and MDA(both P<0.05),and downregulated the protein expression of p-AKT(P<0.05),while its combination with gefitinib had a significantly greater effect(all P<0.05).Conclusion This study shows that saikosaponin e and gefitinib exert a therapeutic effect on cholangiocarcinoma by inhibiting the PI3K-AKT signaling pathway,which provides theoretical support and a scientific basis for further research and clinical application of saikosaponins.关键词
柴胡皂苷/吉非替尼/胆管肿瘤/药理学Key words
Saikosaponins/Gefitinib/Bile Duct Neoplasms/Pharmacology引用本文复制引用
李姗,赵方言,程新月,刘湘花,杨艺,闵远骞,李天星,禄保平..基于网络药理学探讨柴胡皂苷e联合吉非替尼治疗胆管癌的作用机制[J].临床肝胆病杂志,2025,41(12):2586-2596,11.基金项目
河南省高等学校青年骨干教师培养计划(2021GGJS084) （2021GGJS084）
河南省高等学校重点科研项目(21B310003) （21B310003）
河南省中医药科学研究专项课题(2023ZY2110) Training Program for Young Scholars in Universities of Henan Province(2021GGJS084) （2023ZY2110）
Important Scientific Research Project of Universities in Henan Province(21B310003) （21B310003）
Special Scientific Research Project of Chinese Medicine in Henan Province(2023ZY2110) （2023ZY2110）
