基于生物信息学及实验验证探讨麦冬皂苷D治疗特发性肺纤维化的作用机制  

Integrated Bioinformatics and Experimental Validation to Explore the Mechanism of Ophiopogonin D for the Treatment of Idiopathic Pulmonary Fibrosis

作　　者：李仲普[1] 黄乐 刘雨[1] 胡学军[1] 邓秀娟[1] 管聘 LI Zhongpu;HUANG Le;LIU Yu;HU Xuejun;DENG Xiujuan;GUAN Pin(The Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine,Changsha 410006 Hunan,China)

机构地区：[1]湖南省中医药研究院附属医院,湖南长沙410006

出　　处：《中药新药与临床药理》2025年第2期219-229,共11页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(82004306);湖南省中医药科研计划项目(B2023017);湖南省自然科学基金项目(2024JJ5237)。

摘　　要：目的整合生物信息学、网络药理学及分子对接技术预测麦冬皂苷D(OP-D)治疗特发性肺纤维化(IPF)的作用机制,并进行细胞实验验证。方法(1)利用R软件的Limma包分析GEO数据库中IPF差异表达基因数据,筛选IPF疾病靶点;采用PharmMapper和SwissTargetPrediction数据库预测OP-D作用靶点;对筛选得到的OP-D作用靶点与IPF疾病靶点取交集,得到OP-D治疗IPF的潜在作用靶点。对潜在作用靶点进行GO功能及KEGG通路富集分析;采用STRING数据库和Cytoscape软件构建潜在作用靶点的蛋白互作(PPI)网络,并筛选OP-D抗IPF的核心靶点。使用Autodock Vina软件对OP-D与核心靶点进行分子对接验证。(2)采用TGF-β(10μg·L^(-1))诱导人胚肺成纤维细胞(HFL^(-1)),并用不同浓度OP-D进行干预。采用CCK-8法、EdU法检测细胞增殖情况;Transwell检测细胞迁移能力;TUNEL染色法检测细胞凋亡情况;Western Blot法检测细胞中相关蛋白的表达水平。结果(1)筛选得到IPF疾病靶点2040个,OP-D作用靶点408个,取交集得到30个OP-D治疗IPF的潜在作用靶点。OP-D抗IPF的生物过程主要在胶原蛋白分解代谢过程、胶原蛋白代谢过程、细胞外基质分解和对异生物刺激的反应方面;主要涉及通路包括肥厚型心肌病信号通路、类风湿性关节炎信号通路、松弛素信号通路、PPAR信号通路和IL^(-1)7信号通路等。进一步筛选得到IGF1、MMP1、MMP2、MMP3、MMP7、ACE、CCL5等OP-D抗IPF的核心靶点,分子对接显示OP-D与核心靶点均有较好的结合活性。(2)OP-D能浓度依赖性抑制HFL^(-1)细胞的活力,选择1、2、4μmol·L^(-1)OP-D进行后续实验。与正常对照组比较,TGF-β组细胞的EdU阳性率及迁移细胞数显著升高(P<0.01);TUNEL阳性率及Bax蛋白表达水平显著下降(P<0.01),Bcl-2、FN-1、α-SMA、CollagenⅠ、IGF1、MMP1、MMP2、MMP3、MMP7蛋白表达水平显著升高(P<0.05,P<0.01)。与TGF-β组比较,1、2、4μmol·L^(-1)OP-D组的细胞EdU阳性�Objective To integrate bioinformatics,network pharmacology and molecular docking techniques to predict the mechanism of ophiopogonin D(OP-D)in the treatment of idiopathic pulmonary fibrosis(IPF)and to verify it by cell experiments.Methods(1)The Limma package for R software was used to analyze the IPF gene expression data in the GEO database and to screen the IPF disease targets.PharmMapper and SwissTargetPrediction databases were used to predict potential OP-D targets,and targets of OP-D for the treatment of IPF were screened by taking the intersection of OP-D targets and IPF targets.Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were performed on the target.The STRING database and Cytoscape software were used to construct the protein-protein interaction(PPI)network of potential targets,and the core targets of OP-D for anti-IPF were screened.Autodock Vina software was used to perform molecular docking between OP-D and core targets.(2)TGF-β(10μg·L^(-1))was used to induce human embryonic lung fibroblasts,which were treated with OP-D at different concentration.CCK-8 and EdU methods were applied to detect cell proliferation.Transwell system was employed to determine cell migration.TUNEL staining was used to measure cell apoptosis.Western Blot method was applied to detect the expression levels of related proteins in cells.Results(1)A total of 2040 IPF disease targets and 408 OP-D targets were screened.The intersection of the two were taken to obtain 30 potential targets of OP-D for the treatment of IPF.GO and KEGG analysis showed that the main biological processes of OP-D for the treatment of IPF involved in collagen catabolism,collagen metabolism,extracellular matrix decomposition,and the stimulus of xenobiotics,and the enrichment pathways were mainly involved in hypertrophic cardiomyopathy signaling pathway,rheumatoid arthritis signaling pathway,relaxin signaling pathway,PPAR signaling pathway and IL^(-1)7 signaling pathway.The core targets of OP-D for anti-

关 键 词：麦冬皂苷D 特发性肺纤维化 生物信息学 网络药理学 分子对接 实验验证 人胚肺成纤维细胞 

分 类 号：R285.5[医药卫生—中药学] R857.3[医药卫生—中医学]