基于网络药理学、分子对接及实验探讨茵陈蒿汤调节阻塞性黄疸氧化应激的作用机制  

Based on network pharmacology,molecular docking,and experiments to explore the mechanism of Yinchenhao Decoction in regulating oxidative stress of obstructive jaundice

作　　者：陈帅 刘军舰 尚海涛 张井虹 李忠廉 CHEN Shuai;LIU Jun-jian;SHANG Hai-tao;ZHANG Jing-hong;LI Zhong-lian(Graduate School of Tianjin Medical University,Tianjin 300070,China;Department of The Second Hepatobiliary and Pancreatic Surgery,Tianjin Hospital of Integrated Traditional Chinese and Western Medicine,Tianjin 300100,China)

机构地区：[1]天津医科大学研究生院,天津300070 [2]天津市中西医结合医院肝胆胰第二外科,天津300100

出　　处：《天津医科大学学报》2021年第6期595-602,共8页Journal of Tianjin Medical University

基　　金：国家自然科学基金面上项目(81273952);天津市中医药重点领域科技项目(2019003)。

摘　　要：目的:运用网络药理学探讨茵陈蒿汤调节阻塞性黄疸氧化应激的作用机制,并用分子对接及免疫组化进行验证。方法:首先利用中药系统药理学数据库和分析平台(TCMSP)和UniProt数据库检索并筛选茵陈蒿汤有效成分及作用靶点;通过Gene Cards数据库检索阻塞性黄疸的作用靶点;利用R语言将疾病与药物的有效靶点进行分析,确定交集靶点;通过STRING数据库和Cytoscape软件构建蛋白互作网络;利用R语言进行基因本体论(GO)生物学过程富集分析和京都基因与基因组百科全书(KEGG)通路富集分析;利用Autodock_vina软件对内皮型一氧化氮合成酶(eNOS)、诱导型一氧化氮合成酶(iNOS)和茵陈蒿汤主要化学成分进行分子对接验证;最后通过免疫组织化学方法(IHC)验证茵陈蒿汤对阻塞性黄疸大鼠肝组织中eNOS和iNOS的影响。结果:研究共筛选获得茵陈蒿汤中29个有效成分及177个作用靶点,阻塞性黄疸的2183个疾病靶点,茵陈蒿汤和阻塞性黄疸的交集靶点123个;经GO和KEGG富集分析得到133个GO生物过程和127个相关信号通路,主要涉及Hepatitis B、流体剪应力与动脉粥样硬化、丝裂原活化蛋白激酶(MAPK)、磷脂酰肌醇3激酶(PI3K)-蛋白激酶B(Akt)、肝细胞癌、低氧诱导因子(HIF)-1等信号通路;拓扑属性分析发现槲皮素、山奈酚、异鼠李素等主要成分和Akt1、TP53、白细胞介素(IL)6、血管内皮生长因子(VEGF)A、eNOS和iNOS等关键靶点。分子对接验证表明,eNOS和iNOS蛋白与槲皮素、山奈酚、异鼠李素有较好的亲和能力;IHC实验证明与假手术组相比,模型组肝组织中eNOS表达水平降低,而iNOS表达水平升高(P<0.05);与模型组相比,茵陈蒿汤组肝组织中eNOS表达水平升高,iNOS表达水平降低(P<0.01)。结论:茵陈蒿汤对阻塞性黄疸的作用具有多成分、多靶点、多通路的特点,茵陈蒿汤对阻塞性黄疸氧化应激的调节可能与改变eNOS、iNOS蛋白的表达有Objective:To explore the mechanism of Yinchenhao Decoction in regulating oxidative stress of obstructive jaundice by network pharmacology,and to verify it by molecular docking and immunohistochemistry.Methods:TCMSP and UniProt database were used to search and screen the effective components and targets of Yinchenhao Decoction.Gene Cards database was used to search the targets of obstructive jaundice.R language was used to analyze the effective targets of diseases and drugs to determine the intersection targets.STRING database and Cytoscape software were used to construct Protein Interaction network.R language was used to analyze the biological process of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.Autodock_Vina software was used to verify the molecular docking of endothelial nitric oxide synthase(eNOS),inducible nitric oxide synthase(iNOS),and main chemical components in Yinchenhao Decoction.Finally,the effect of Yinchenhao Decoction on eNOS and iNOS in liver tissue of rats with obstructive jaundice was verified by immunohistochemistry(IHC).Results:A total of 29 active components and 177 targets in Yinchenhao Decoction,2183 disease targets in obstructive jaundice and 123 intersection targets in Yinchenhao Decoction and obstructive jaundice were screened.Through GO and KEGG enrichment analysis,133 GO biological processes and 127 related signaling pathways were obtained,mainly involving the signaling pathways of hepatotis B,fluid shear stress and atherosclerosis,MAPK,PI3K-Akt,hepatocellular carcinoma,HIF-1 and so on.The main components of the content of Quercetin,Kaempferol and Isorhamnetin and the key targets of Akt1,TP53,IL6,VEGFA,eNOS and iNOS were found by Topological Analysis.Molecular docking showed that eNOS and iNOS proteins had good affinity with Quercetin,Kaempferol and Isorhamnetin.IHC experiment showed that compared with sham operation group,the expression level of eNOS in model group was lower(P<0.05);compared with model group,the expression level of eN

关 键 词：茵陈蒿汤 阻塞性黄疸 网络药理学 分子对接 ENOS INOS 

分 类 号：R285[医药卫生—中药学]