基于网络药理学和实验验证探...炎小鼠的治疗作用及作用机制_张永康.pdf

中草药 中草药 2023 年年 3 月月 第第 54 卷卷 第第 5 期期 Chinese Traditional and Herbal Drugs 2023 March Vol.54 No.5 1461 基于网络药理学和实验验证探讨荆防颗粒对自身免疫性肝炎小鼠的治疗作用及作用机制 张永康1,2，孙成宏2,3，王西双2,3，姚景春2,3，张贵民1,2*1.山东中医药大学药学院，山东 济南 250300 2.鲁南制药集团股份有限公司 中药制药共性技术国家重点实验室，山东 临沂 276006 3.鲁南制药集团股份有限公司 临沂市天然药物免疫药理毒理重点实验室，山东 临沂 276006 摘 要：目的 基于网络药理学及动物实验探讨荆防颗粒对自身免疫性肝炎（autoimmune hepatitis，AIH）小鼠的治疗作用及作用机制。方法 通过 TCMSP 数据库筛选荆防颗粒的活性成分及其对应的靶点，通过检索 Omim、Drugbank 和 GeneCards 数据库收集 AIH 相关的靶点，进而得到荆防颗粒治疗 AIH 的关键靶点；将获得的关键靶点导入 STRING 数据库进行分析，并构建蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络，通过 Cytoscape 软件可视化；通过 Metascape 数据库对关键靶点进行基因本体（gene ontology，GO）功能及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析。通过建立刀豆蛋白 A 诱导的 AIH 小鼠模型探讨荆防颗粒治疗 AIH 的作用机制。结果 共筛选得到荆防颗粒中159 个潜在活性成分和 269 个相关的靶点，与 343 个 AIH 相关靶点取交集，获得 25 个交集靶点。PPI 网络分析显示，白细胞介素-6（interleukin-6，IL-6）、肿瘤坏死因子-（tumor necrosis factor-，TNF-）、IL-1、信号传导和转录激活蛋白 3（signal transducer and activator of transcription 3，STAT3）和 IL-8/CXC 型趋化因子配体 8（CXC chemokine ligand 8，CXCL8）等靶点可能为荆防颗粒治疗 AIH 的关键靶点；KEGG 通路富集分析得到炎症途径和凋亡相关途径信号通路。体内实验结果显示，与模型组比较，荆防颗粒显著减轻了刀豆蛋白 A 诱导的肝炎，表现为小鼠存活率增加、肝细胞坏死减少、血清中丙氨酸氨基转移酶（alanine aminotransferase，ALT）和天冬氨酸氨基转移酶（aspartate aminotransferase，AST）活性降低（P0.05、0.01）；荆防颗粒还通过抑制 IL-6/STAT3、NOD 样受体热蛋白结构域相关蛋白 3（NOD-like receptor thermal protein domain associated protein 3，NLRP3）/IL-1 和 TNF-/核因子-B（nuclear factor-B，NF-B）通路进而调节多种细胞因子（IL-6、IL-1、TNF-、CXCL-8）的产生（P0.05、0.01），从而发挥抗炎、抗凋亡作用。结论 荆防颗粒通过多成分、多靶点发挥治疗 AIH 的作用。关键词：荆防颗粒；自身免疫性肝炎；网络药理学；炎症因子；IL-6/STAT3 信号通路；NLRP3/IL-1 信号通路；TNF-/NF-B 信号通路 中图分类号：R285.5 文献标志码：A 文章编号：0253-2670(2023)05-1461-10 DOI:10.7501/j.issn.0253-2670.2023.05.012 Therapeutic effect and mechanism of Jingfang Granules on autoimmune hepatitis mice based on network pharmacology and experimental verification ZHANG Yong-kang1,2,SUN Cheng-hong2,3,WANG Xi-shuang2,3,YAO Jing-chun2,3,ZHANG Gui-min1,2 1.College of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250300,China 2.State Key Laboratory of Generic Manufacture Technology of Chinese Traditional Medicine,Lunan Pharmaceutical Group Co.,Ltd.,Linyi 276006,China 3.Linyi Key Laboratory for Immunopharmacology and Immunotoxicology of Natural Medicine,Lunan Pharmaceutical Group Co.Ltd.,Linyi 276006,China Abstract:Objective To explore the therapeutic effect and mechanism of Jingfang Granules(荆防颗粒)on autoimmune hepatitis(AIH)mice based on network pharmacology and animal experiments.Methods The active components of Jingfang Granules and corresponding targets were screened by TCMSP database,and targets related to AIH were collected by Omim,Drugbank and 收稿日期：2022-12-26 基金项目：国家“重大新药创制”科技重大专项（2021CXGC010508）作者简介：张永康（1997），男，硕士，主要从事中药药理学研究。E-mail:*通信作者：张贵民，研究员，主要从事创新药物研发。E-mail: 1462 中草药 中草药 2023 年年 3 月月 第第 54 卷卷 第第 5 期期 Chinese Traditional and Herbal Drugs 2023 March Vol.54 No.5 GeneCards databases,and then the key targets of Jingfang Granules in treating AIH were obtained.The obtained key targets are imported into STRING database for analysis,and protein-protein interaction(PPI)network was constructed and visualized by Cytoscape software.Gene ontology(GO)function and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis of key targets were analyzed by Metascape database.AIH mice model induced by concanavalin A was established to explore the mechanism of Jingfang Granules in treating AIH.Results A total of 159 potential active components and 269 related targets in Jingfang Granules were screened,and 25 intersecting targets were obtained by intersecting with 343 AIH related targets.PPI network analysis showed that interleukin-6(IL-6),tumor necrosis factor-(TNF-),IL-1,signal transducer and activator of transcription 3(STAT3)and IL-8/CXC chemokine ligand 8(CXCL8)may be the key targets of Jingfang Granules in treating AIH.KEGG pathway enrichment analysis obtained inflammatory pathway and apoptosis-related pathway signal pathway.The experimental results in vivo showed that compared with model group,Jingfang Granules significantly reduced the hepatitis induced by concanavalin A,which showed that survival rate of mice was increased,necrosis of hepatocytes was decreased,activities of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum were decreased(P 0.05,0.01).Jingfang Granules regulated the production of multiple cytokines including IL-6,IL-1,TNF-and CXCL-8 by inhibiting IL-6/STAT3,NOD-like receptor thermal protein domain associated protein 3(NLRP3)/IL-1 and TNF-/nuclear factor-B(NF-B)pathways(P 0.05,0.01),thereby playing an anti-inflammatory and anti-apoptosis role.Conclusion Jingfang Granules plays a role in treating AIH through multi-components and multi-targets.Key words:Jingfang Granules;autoimmune hepatitis;network pharmacology;inflammatory factors;IL-6/STAT3 signal pathway;NLRP3/IL-1 signal pathway;TNF-/NF-B signal pathway自身免疫性肝炎（autoimmune hepatitis，AIH）是一种以肝实质损害为特征的炎症性疾病，在亚太地区的发病率约为 0.005%0.025%，且呈逐年上升趋势1，如果不及时治疗，则会迅速发展为肝硬化和肝衰竭，严重危害人类健康。AIH 的发病原因较为复杂，临床上多以糖皮质类激素如地塞米松磷酸钠注射液等进行治疗，但极易引起机体诸多不良反应2。因