基于网络药理学方法和细胞实验探讨长春布宁治疗肝纤维化的作用机制  

Mechanism of vinburnine in the treatment of liver fibrosis based upon network pharmacology and cellular experiments

作　　者：宛博 单玲玲[1] 田晓雨 隆毅[2] WAN Bo;SHAN Lingling;TIAN Xiaoyu;LONG Yi(Department of Pharmacy,Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Shandong Jinan 250002,China;Traditional Chinese Medicine Data Center,Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Shandong Jinan 250002,China;Department of Clinical Pharmacy,Qilu Hospital of Shandong University,Shandong Jinan 250014,China)

机构地区：[1]山东中医药大学第二附属医院药学部,山东济南250002 [2]山东中医药大学第二附属医院中医药数据中心,山东济南250002 [3]山东大学齐鲁医院临床药学部,山东济南250014

出　　处：《中国医院药学杂志》2024年第22期2557-2562,共6页Chinese Journal of Hospital Pharmacy

基　　金：山东省药品临床综合评价项目(编号:2022YZ008);山东省中医药科技项目(编号:Z-2023055);山东省中医药科技项目(编号:M-2023169);全国名老中医药专家传承工作室建设项目(编号:国中医药人教函[2022]75号)。

摘　　要：目的:利用网络药理学和体外细胞实验验证的方法探讨长春布宁治疗肝纤维化的潜在作用机制。方法:通过检索Super-PRED、Similarityensembleapproach(SEA)、BindingDB、ChEMBL、PharmMapper、SwissTargetPrediction以及BATMAN-TCM数据库得到长春布宁的相关作用靶点,并与检索DisGeNET、OMIM和MalaCards数据库得到的肝纤维化作用靶点取交集,获得长春布宁治疗肝纤维化潜在靶点。利用STRING数据库构建上述靶点间的蛋白质相互作用网络,使用Cytoscape 3.9.0软件进行可视化;再利用Metascape平台对其进行功能和通路富集分析。在体外实验中,利用TGFβ1诱导人肝星状细胞LX-2活化,体外模拟肝纤维化的发生;再给予长春布宁处理,通过双荧光素酶报告基因实验,Western blot,realtime PCR等方法评价其抗肝纤维化活性。结果:通过筛选匹配后获得长春布宁治疗肝纤维化潜在靶点共计140个,其中核心靶点包括BCL2L1、CASP3、MAPK1、STAT3、EGFR、HIF1A、PTGS2、MMP9等;GO、KEGG通路以及Reactome通路分析显示长春布宁可能通过影响细胞迁移、凋亡、炎症反应、创伤反应等生物学过程调控肝纤维化,并且涉及PI3K-AKT、MAPK、HIF等细胞信号通路。体外实验中,长春布宁呈剂量依赖性地抑制LX-2细胞内COL1A1以及TGFB1的启动子活性;Western blot和real-time PCR检测结果表明长春布宁对LX-2细胞中纤维化标志物的表达水平具有显著抑制作用。结论:本研究初步验证长春布宁有望成为治疗肝纤维化的潜在化合物,并探讨其可能的作用机制,为后续研究提供实验基础。OBJECTIVE To explore the potential mechanism of vinbumine intervening liver fibrosis(LF)using network pharmacology and in vitro experimental validation.METHODS The relevant targets of vinburnine were obtained from the data-bases of Super-PRED,Similarity ensemble approach(SEA),BindingDB,ChEMBL,PharmMapper,Swiss Target Prediction and BATMAN-TCM.The targets of LF were acquired from the databases of DisGeNET,OMIM and MalaCards.The intersec-tions of the above two were the potential therapeutic targets of vinbumine for LF.The protein-protein interaction network of these targets was constructed by the database of STRING and visualized by Cytoscape 3.9.0 software.Functional and pathway enrich-ment analyses were performed with the platform of Metascape.Human hepatic stellate cell line,LX-2,was applied in in vitro experiments.TGFβ1 was utilized for inducing the activation of LX-2 cells,which simulates the occurrence of LF.Anti-fibrotic activity of vinburnine was evaluated by dual-luciferase reporter assay,Western blot and real-time polymerase chain reaction(PCR).RESULTS A total of 140 potential therapeutic targets of vinburnine for LF were obtained after screening and matching.Among them,the core targets included BCL2L1,CASP3,MAPK1,STAT3,EGFR,HIF1A,PTGS2 and MMP9,etc.The results of GO/KEGG and Reactome pathway analyses indicated that vinburnine could affect cellular migration,apoptosis,inflam-mation,cellular function,cellular regeneration and cellular regeneration.GO/KEGG and Reactome pathway analyses revealed that vinbumine could regulate LF through affecting cellular migration,apoptosis,inflammatory response and response to wound-ing involved in PI3K-AKT,MAPK and HIF signaling.In LX-2 cells,vinburnine dose-dependently suppressed the promoter activity of COL1A1 and TGFB1.It also exhibited a significant inhibitory effect on the expression of fibrotic markers.CONCLUSION The present study preliminarily demonstrated that vinbumine is a potential therapeutic compound for LF.And elucidating its possible mechanisms shall provid

关 键 词：网络药理学 长春布宁 肝纤维化 肝星状细胞 信号通路 

分 类 号：R96[医药卫生—药理学]