基于网络药理学霍山石斛治疗肝损伤作用机制探讨及体内实验验证  

Mechanism of Dendrobium huoshanense in treatment of liver injury:Based on network pharmacology and in vivo experimental

作　　者：林彩霞 陈涛 王成 贾婉秋 廖茂梁 韩邦兴[2,4] 顾欣雨 袁秀珍 刘东 苏婷[2,4] LIN Caixia;CHEN Tao;WANG Cheng;JIA Wanqiu;LIAO Maoliang;HAN Bangxing;GU Xinyu;YUAN Xiuzhen;LIU Dong;SU Ting(School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012,China;College of Biological and Pharmaceutical Engineering,West Anhui University,Lu'an 237012,China;Huo Shan RainiGrace Biological Technology Development Co.,Ltd.,Lu'an 237012,China;Anhui Engineering Research Center for Eco-agriculture of Traditional Chinese Medicine,Lu'an 237012,China;Anhui Modern Chinese Medicine Industry Common Technology Research Center,Lu'an 237012,China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230012 [2]皖西学院生物与制药工程学院,安徽六安237012 [3]霍山县天下泽雨生物科技发展有限公司,安徽六安237012 [4]安徽省中药生态农业工程研究中心,安徽六安237012 [5]安徽省现代中药产业共性技术研究中心,安徽六安237012

出　　处：《药物评价研究》2024年第7期1502-1512,共11页Drug Evaluation Research

基　　金：国家十四五重点研发计划项目(2023YFC3503804);安徽省科技重大专项(202103b06020004);安徽省大别山中医药研究院开放课题重点科技攻关项目(TCMADM-2023-02,TCMADM-2023-08);安徽省中药生态农业工程研究中心开放课题(WXZR202316);皖西学院高层次人才基金项目(WGKQ2021083,WGKQ2022074);国家级、安徽省大学生创新创业训练项目(202310376037,s202310376123);安徽省中药资源保护与持续利用工程实验室开放课题(TCMRPSU-2022-01)。

摘　　要：目的基于网络药理学、分子对接及体内实验方法探讨霍山石斛对肝损伤的治疗作用及其机制。方法运用中国学术期刊全文数据库(CNKI)、PubChem等数据库获取霍山石斛成分,通过GeneCards数据库筛选肝损伤相关基因,将成分靶点与疾病基因交集后构建蛋白质-蛋白质相互作用(PPI)网络图,进行基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)通路富集分析,Cytoscape构建“药物-化合物-靶点”网络图,利用AutoDock Tools对关键活性成分和核心靶点进行分子对接。构建乙醇诱导的肝损伤小鼠模型,通过观察肝脏病理学改变,试剂盒法测定血清中天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)及白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平,通过实时荧光定量逆转录PCR(qRT-PCR)检测酒精性肝损伤小鼠肝脏中相关基因的表达,考察霍山石斛对酒精性肝损伤小鼠的影响。结果筛选出霍山石斛中33个活性成分,与肝损伤共有靶点224个,包含AKT1、EGFR、SRC等,涉及癌症途径、PI3K/Akt通路等。体内实验结果显示,霍山石斛水提物能够缓解小鼠酒精性肝损伤,并极显著降低AST、ALT和TNF-α、IL-6水平(P<0.01),极显著下调IκB激酶(IKK)、核因子-κB(NF-κB)、蛋白激酶B(Akt)mRNA表达水平(P<0.01)。结论网络药理学分析及动物实验结果,初步证明霍山石斛水提物可以通过下调Akt、IKK、NF-κB mRNA表达水平,影响NF-κB信号通路上游,参与酒精性肝损伤通路中炎症反应的调控,提示霍山石斛具有改善酒精性肝病的潜在作用,为揭示霍山石斛治疗肝损伤作用机制提供参考。Objective To explore the efficacy and mechanism of action of Dendrobium huoshanense in treating liver injury using network pharmacology,molecular docking,and in vivo experiments.Methods China National Knowledge Infrastructure(CNKI),PubChem,and other databases were utilized to identify the components of D.huoshanense.Treatment-related genes for liver injury were selected through the GeneCards databases.After intersecting the component targets with disease genes,a protein-protein interaction(PPI)network was constructed.Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis was performed.A"Drug-Compound-Target"network was established using Cytoscape,and key active components and core targets were docked using AutoDock Tools.An ethanol-induced liver injury mouse model was developed.Liver pathology sections were observed,and levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in serum were measured.The expression of related genes in the liver of alcoholic liver injury mice treated with an aqueous extract of D.huoshanense was detected through realtime fluorescence quantitative reverse transcription PCR(qRT-PCR)to assess the impact on alcoholic liver injury mice mice.Results A total of 33 active components were identified in D.huoshanense,correlating with 224 common targets for liver injury,including Akt,EGFR,SRC,and others,involving pathways like cancer and the PI3K/Akt pathway.In vivo experiments showed that the aqueous extract of D.huoshanense could alleviate liver injury in alcoholic liver injury mice.It significantly reduced levels of ALT,AST,TNF-α,and IL-6(P<0.01)and markedly downregulated the expression of IκB kinase(IKK),nuclear factor-κB(NF-κB)and protein kinase B(Akt)(P<0.01).Conclusion Network pharmacology analysis and animal experimental results preliminary demonstrate that the aqueous extract of D.huoshanense can modulate the inflammatory response in the alcohol liver di

关 键 词：霍山石斛 肝损伤 网络药理学 IκB激酶(IKK) PI3K/Akt/NF-κB信号通路 

分 类 号：R285.5[医药卫生—中药学]