基于网络药理学和实验验证探讨陈皮藿香汤治疗腹泻的作用机制  

Mechanism of Chenpi Huoxiangtang Against Diarrhea:Based on Network Pharmacology and Experimental Verification

作　　者：李哲[1] 李娜[2] 崔晓燕 赵蓝青青 于永洲 赵春颖[1] 熊辉 LI Zhe;LI Na;CUI Xiaoyan;ZHAO-LAN Qingqing;YU Yongzhou;ZHAO Chunying;XIONG Hui(Hebei Key Laboratory of Study and Exploitation of Chinese Medicine,Chengde Medical University,Chengde 067000,China;Hebei Key Laboratory of Nerve Injury and Repair,Chengde Medical University,Chengde 067000,China;Hebei Institute of Drug and Medical Device Inspection,Shijiazhuang 050011,China;Institute of Basic Medical Research,Chengde Medical University,Chengde 067000,China)

机构地区：[1]承德医学院河北省中药研究与开发重点实验室,河北承德067000 [2]承德医学院河北省神经损伤与修复重点实验室,河北承德067000 [3]河北省药品医疗器械检验研究院,石家庄050011 [4]承德医学院基础医学研究所,河北承德067000

出　　处：《中国实验方剂学杂志》2022年第23期79-86,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年科学基金项目(82104384);河北省高等学校科学技术研究项目(QN2021008);承德医学院高层次人才科研启动项目(202103);承德医学院中药药效物质基础青年PI科技创新团队;中央引导地方科技发展资金项目(216Z2501G);河北省高校重点学科建设项目(冀教高[2013]4);河北省科技厅“技术创新引导专项-科技工作会商”项目。

摘　　要：目的:基于网络药理学和实验验证探讨陈皮藿香汤治疗腹泻的作用机制。方法:通过中药系统药理学数据库与分析平台(TCMSP)筛选出陈皮藿香汤的活性成分及作用靶点。采用GeneCards及在线人类孟德尔遗传数据库(OMIM)检索腹泻疾病相关靶点,聚焦共同靶标,借助Cytoscape 3.7.1及STRING数据库构建成分-靶点-疾病及蛋白质-蛋白质相互作用(PPI)网络,采用Bioconductor平台和R语言对核心靶标进行基因本体(GO)富集和京都基因与基因组百科全书(KEGG)通路分析。利用AutoDockTools 1.5.6和AutoDockVina 1.1.2软件中进行分子对接。最后通过动物试验进一步验证实验结果。结果:陈皮藿香汤的主要活性成分51个、对应靶点226个,药物疾病共同靶点37个;GO富集分析得1 249个条目(P<0.05)、KEGG通路富集分析筛选出110条信号通路(P<0.05),主要涉及磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)、Janus激酶/信号转导子与转录激活因子(JAK/STAT)、缺氧诱导因子-1(HIF-1)、丝裂原活化蛋白激酶(MAPK)等信号通路。分子对接结果表明橙皮苷与Akt1结合能力最强。动物实验结果显示大鼠陈皮藿香汤给药后可以显著缓解大鼠的腹泻症状,给药第1天腹泻指数降为0.92,与模型组腹泻指数(1.24)差异有明显统计学意义(P<0.05),小肠墨汁推进率70.36%,经统计学分析与模型组(58.20%)差异有显著统计学意义(P<0.01),给药后可改善大鼠的肠蠕动功能。酶联免疫吸附测定法(ELISA)检测结果显示白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)水平表达明显降低(P<0.05),Akt1、表皮细胞生长因子(EGF)、表皮生长因子受体(EGFR)水平表达明显升高(P<0.05)。结论:陈皮藿香汤中的有效成分主要通过Akt1、IL-6、IL-1β、EGF、EGFR等核心靶点调节PI3K/Akt、MAPK等信号通路发挥对腹泻的治疗作用,为后续研究陈皮-藿香药对药理机制,两者间配伍的科学内涵提供了实验依据。Objective:To explore the mechanism of Chenpi Huoxiangtang against diarrhea based on network pharmacology and experimental verification.Method:The active components and targets of Chenpi Huoxiangtang were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),and targets related to diarrhea from GeneCards and Online Mendelian Inheritance in Man(OMIM).Thereby,the common targets were screened out.The component-target-disease network and proteinprotein interaction(PPI)network were constructed with Cytoscape 3.7.1 and STRING.Bioconductor and R language were employed for gene ontology(GO)term enrichment and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment of core targets,and AutoDockTools 1.5.6 and AutoDockVina 1.1.2 for molecular docking.Finally,animal experiment was carried out for verification.Result:A total of 51 active components 226 targets of Chenpi Huoxiangtang and 37 common targets of the medicine and the disease were screened out.The core targets were involved in 1 249 GO terms(P<0.05)and 110 KEGG pathways(P<0.05),mainly the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt),Janus kinase(JAK)/signal transducer and activator of transcription(STAT),hypoxia inducible factor(HIF)-1,and mitogen-activated protein kinase(MAPK)pathways.Molecular docking suggested that hesperidin and Akt1 showed the highest binding affinity.Animal experiment demonstrated that Chenpi Huoxiangtang significantly alleviated the diarrhea symptoms.The diarrhea index on the first day of administration decreased to 0.92,which was statistically different from that(1.24)of the model group(P<0.05).The intestinal ink propulsion rate was 70.36%,higher than that(58.20%)in the model group(P<0.01).Moreover,intestinal peristalsis was improved by the decoction.Enzyme-linked immunosorbent assay(ELISA)indicated the decrease in the levels of interleukin-6(IL-6)and interleukin-1β(IL-1β)(P<0.05)and increase in the levels of Akt1,epidermal growth factor(EGF),and epidermal growth factor r

关 键 词：网络药理学 实验验证 陈皮藿香汤 腹泻 作用机制 

分 类 号：R284.2[医药卫生—中药学] R285[医药卫生—中医学] R289R22R2-031R33