基于网络药理学及实验验证探讨半枝莲-党参药对治疗膀胱癌的作用机制

Mechanism of action of Scutellaria barbata combined with Codonopsis pilosula in the treatment of Bladder carcinoma based on network pharmacology and experimental validation

目的通过网络药理学研究半枝莲-党参药对治疗膀胱癌的作用机制。方法查询中药系统药理学数据库分析平台(TCMSP)筛选半枝莲-党参药对的药物成分,使用Swiss Target Prediction预测药物成分的作用靶点;运用GeneCards获取膀胱癌的疾病靶点,利用venny 2.1获取交集靶点。使用STRING进行蛋白-蛋白相互作用分析并使用Cytoscape构建"药物-靶点-通路"网络,利用Metascape进行基因本体(GO)和京都基因与基因组百科全书(KEGG)分析。将裸鼠随机分为模型组和治疗组,建立膀胱癌小鼠模型。模型组小鼠灌胃等剂量溶剂,治疗组建模后第8天灌胃半枝莲-党参药对0.2 ml,剂量为342.86 mg/kg,2次/d。建模后第28天,测量裸鼠瘤体大小,并采用酶联免疫吸附试验法检测前列腺素G/H合成酶2(PTGS2)、PTGS1、核受体辅活化子2(NCOA2)、维甲酸X受体α(RXRA)、孕酮受体(PGR)、丝裂原活化蛋白激酶1(MAPK1)、网状内皮增生病毒癌基因同源物A(RELA)、蛋白激酶B1(Akt1)水平。结果半枝莲-党参药对的45个药物成分通过多条通路直接作用于187个疾病靶点治疗膀胱癌,主要核心成分为槲皮素、木犀草素、汉黄芩素、7-甲氧基-2-甲基异黄酮、黄芩素、β-谷甾醇和豆甾醇等,关键靶点为PTGS2、PTGS1、NCOA2、RXRA、PGR、MAPK1、RELA和Akt1等。GO富集分析显示,生物过程主要涉及对激素的反应、细胞对脂质的反应、对外来刺激的反应、对细菌分子的反应等,细胞组分主要涉及转录调控复合体、膜筏、膜微区、RNA聚合酶Ⅱ转录调控复合物等,分子功能主要涉及转录因子结合、DNA结合转录因子结合、RNA聚合酶Ⅱ特异性DNA结合转录因子结合、核受体活性、配体激活的转录因子活性等。KEGG通路富集分析显示半枝莲-党参药对治疗膀胱癌的主要信号通路为晚期糖基化终产物及其受体(AGE-RAGE)、白细胞介素-17(IL-17)、磷脂酰肌醇-3-激酶-蛋白激酶B(PI3K-Akt)、肿瘤坏死因子(TNF)、MAPK、缺氧诱导因子-1(HIF-1)、细胞凋亡、p53、Toll样受体等。动物实验验证显示,半枝莲-党参药对能够明显改善裸鼠的瘤体大小,同时也能改善PTGS2、PTGS1、NCOA2、RXRA、PGR、MAPK1、RELA、Akt1表达水平。结论半枝莲-党参药对主要通过调节AGE-RAGE、IL-17、PI3K-Akt、TNF、MAPK、HIF-1、细胞凋亡、p53、Toll样受体等信号通路的PTGS2、PTGS1、NCOA2、RXRA、PGR、MAPK1、RELA、Akt1等靶点治疗膀胱癌。

ObjectiveTo investigate the mechanism of bladder cancer treatment by using Scutellaria barbata and Codonopsis pilosula drug pair through network pharmacology.MethodsThe drug composition of the drug pair was screened using TCMSP,and their action targets were predicted using Swiss Target Prediction.GeneCards was used to obtain disease targets of bladder cancer,and venny 2.1 was used to obtain intersection targets.PPI analysis was performed using STRING,and a network diagram was constructed using Cytoscape.GO and KEGG analysis were conducted using Metascape.A drug-target-pathway network map was constructed using Cytoscape software.Nude mice were randomly divided into a model group and a treatment group to establish a bladder cancer mouse model.On the 8th day after model formation,the mice in the model group were administered intragastrically with a dose of 342.86 mg/kg,0.2 ml,twice/day.On the 28th day after modeling,the tumor size of nude mice was measured.Prostaglandin G/H Synthetase 2(PTGS2),PTGS1,Nuclear Receptor Coactivator 2(NCOA2),Retinoic Acid X Receptorα(RXRA),Progesterone Receptor(PGR),Mitogen-Activated Protein Kinase 1(MAPK1),Reticuloendothelial Proliferation virus oncogene homology A(RELA),and Akt1 levels were detected by enzyme-linked immunosorbent assay.ResultsThe results show that 45 active components of the drug pair directly acted on 187 disease targets through multiple pathways to treat bladder cancer,in which Quercetin,luteolin,wogonin,7-Methoxy-2-methyl isoflavone,baicalein,beta-sitosterol,Stigmastero,and other core ingredients,as well as PTGS2,PTGS1,NCOA2,RXRA,PGR,MAPK1,RELA,and Akt1 are critical targets.The results of gene function annotation analysis show that the biological processes most likely related to crossover genes mainly involved responses to hormones,cell responses to lipids,responses to foreign stimuli,and responses to bacterial molecules.The cell components mainly involves transcription regulatory complexes,membrane rafts,membrane microregions,and RNA polymeraseⅡtranscriptional regulatory complexes,etc.The molecular functions mainly involve transcription factor binding,DNA-binding transcription factor binding,RNA polymeraseⅡspecific DNA-binding transcription factor binding,nuclear receptor activity,ligand-activated transcription factor activity,etc.The results of pathway enrichment analysis suggests that the main signaling pathways are AGE-RAGE,IL-17,PI3K-Akt,TNF,MAPK,HIF-1,apoptosis,p53,toll-like receptor,etc.Animal experiments show that the Scutellaria barbata and Codonopsis pilosula drug pair can significantly improve tumor size and also improve the expression levels of PTGS2,PTGS1,NCOA2,RXRA,PGR,MAPK1,RELA,and Akt1.ConclusionsThe Scutellaria barbata and Codonopsis pilosula drug pair can regulate PTGS2,PTGS1,NCOA2,RXRA,PGR,MAPK1,RELA,and Akt1 and other diseases mainly through the regulation of AGE-RAGE,IL-17,PI3K-Akt,TNF,MAPK,HIF-1,apoptosis,p53,toll-like receptor,and other signaling pathways.Targeting enzyme activity and cell apoptosis can treat bladder cancer by regulating these biological processes.