中药新药与临床药理2025，Vol.36Issue(9)：1492-1506,15.DOI:10.19378/j.issn.1003-9783.2025.09.009
基于网络药理学及实验验证探讨健脾清热活血方治疗胃癌前病变的作用机制
Exploring the Mechanism of Jianpi Qingre Huoxue Formula in Treating Precancerous Lesions of Gastric Cancer Through Network Pharmacology and Experimental Validation
摘要
Abstract
Objective To investigate the potential mechanism of Jianpi Qingre Huoxue Formula(JQHF)in treating precancerous lesions of gastric cancer(PLGC)using network pharmacology and experimental validation.Methods(1)Active components of JQHF and their corresponding targets were screened through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Herb database,and related literature.PLGC-related disease targets were identified from GeneCards and OMIM databases.The intersection targets between JQHF and PLGC were obtained to identify potential therapeutic targets.A protein-protein interaction(PPI)network was constructed using the STRING database,and a"disease-drug-active component-target"network was established using Cytoscape software to screen core active components and potential targets of JQHF for PLGC treatment.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed on potential targets using Metascape software,and molecular docking was used to validate the binding activity between core active components and potential targets.(2)A malignant transformation cell(MC)model of PLGC was established by inducing GES-1 cells with 1-methyl-3-nitro-1-nitrosoguanidine(MNNG).Blank serum and drug-containing serum of JQHF were prepared from SD rats,and the optimal intervention concentration and time of JQHF drug-containing serum were determined using the CCK-8 assay.Subsequently,GES-1 cells in the logarithmic growth phase were randomly divided into a blank group(20%blank serum),model group(20%blank serum),JQHF low-dose group(10%blank serum+10%JQHF drug-containing serum),and JQHF high-dose group(20%JQHF drug-containing serum).Cell proliferation,migration,and invasion abilities were evaluated using colony formation,scratch,and Transwell assays.The mRNA expression levels of E-cadherin,N-cadherin,Vimentin,caudal-type homeobox transcription factor 2(CDX2),sex-determining region Y-box protein 2(SOX2),tumor protein P53(TP53),serine/threonine-protein kinase 1(AKT1),signal transducer and activator of transcription 3(STAT3),sarcoma viral oncogene homolog(SRC),and heat shock protein 90 α family class A member 1(HSP90AA1)were detected by qRT-PCR.The protein expression levels of E-cadherin,N-cadherin,Vimentin,CDX2,and SOX2 were measured by Western Blot.Results(1)A total of 94 active components of JQHF and 747 corresponding targets were identified,along with 2 074 PLGC-related disease targets.The intersection targets numbered 329.Core active components of JQHF for PLGC treatment included quercetin,12-senecioyl-2E,8E,10E-atractylentriol,apigenin,luteolin,and ursolic acid,while core potential targets included TP53,AKT1,STAT3,SRC,and HSP90AA1.The therapeutic mechanisms primarily involved the PI3K-AKT,TNF,and IL-17 signaling pathways.Molecular docking confirmed strong binding activity between core active components(quercetin,12-senecioyl-2E,8E,10E-atractylentriol,apigenin,luteolin,ursolic acid)and core potential targets(TP53,AKT1,STAT3,SRC,HSP90AA1).(2)The CCK-8 assay determined the optimal intervention concentrations as 10%(low dose)and 20%(high dose)JQHF drug-containing serum,with an intervention time of 48 hours.Compared with the blank group,the model group exhibited significantly increased colony formation,migration area,migrated cell count,and invaded cell count(P＜0.01).In contrast,the JQHF low-and high-dose groups showed significant reductions in these parameters(P＜0.05,P＜0.01).Compared with the blank group,the model group displayed significantly decreased mRNA and protein expression levels of E-cadherin,SOX2,and TP53 mRNA(P＜0.01),while the mRNA and protein expression levels of N-cadherin,Vimentin,CDX2,and mRNA expression of AKT1,STAT3,SRC,and HSP90AA1were elevated(P＜0.01).Compared with the model group,the JQHF low-and high-dose groups exhibited increased mRNA and protein expression levels of E-cadherin,and SOX2 and mRNA expression of TP53(P＜0.05,P＜0.01),while the mRNA expression levels of AKT1,STAT3,SRC,mRNA and protein of N-cadherin,Vimentin,and CDX2 were significantly reduced(P＜0.05,P＜0.01).The high-dose JQHF group also showed decreased HSP90AA1 mRNA expression(P＜0.05),while the low-dose group exhibited a non-significant downward trend,with no statistically significant difference(P＞0.05).Conclusion JQHF treats PLGC through multiple components,targets,and pathways.Core active components such as quercetin,12-senecioyl-2E,8E,10E-atractylentriol,apigenin,luteolin,and ursolic acid may regulate core targets(TP53,AKT1,STAT3,SRC,HSP90AA1)and modulate the PI3K-AKT,TNF,and IL-17 signaling pathways to exert therapeutic effects.Cellular experiments confirmed that JQHF drug-containing serum can reverse malignant biological behaviors of MC cells and delay PLGC progression by targeting TP53,AKT1,STAT3,SRC,and HSP90AA1.关键词
胃癌前病变/健脾清热活血方/网络药理学/分子对接/实验验证/恶性转化细胞Key words
precancerous lesions of gastric cancer/Jianpi Qingre Huoxue Formula/network pharmacology/molecular docking/experimental validation/malignant transformation cell分类
医药卫生引用本文复制引用
麦紫涵,谭浩杰,梁水先,陈彦彤,杨泽虹,李培武..基于网络药理学及实验验证探讨健脾清热活血方治疗胃癌前病变的作用机制[J].中药新药与临床药理,2025,36(9):1492-1506,15.基金项目
国家自然科学基金项目(82474404) （82474404）
广东省自然科学基金面上项目(2023A1515011019). （2023A1515011019）
