中药新药与临床药理2025，Vol.36Issue(2)：219-229,11.DOI:10.19378/j.issn.1003-9783.2025.02.008
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
摘要
Abstract
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(10 μg·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 2 040 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-17 signaling pathway.The core targets of OP-D for anti-IPF were IGF-1,MMP1,MMP2,MMP3,MMP7,ACE and CCL5.Molecular docking indicated that OP-D had good binding activity with core targets.(2)OP-D can concentration-dependently inhibit the viability of HFL-1 cells,and 1,2,and 4 μmol·L-1 OP-D were selected for subsequent experiments.Compared with the normal control group,the EdU positive rate and number of cell migration in TGF-β group were significantly increased(P<0.01).TUNEL positive rate and Bax protein expression level were significantly decreased(P<0.01).The protein expression levels of Bcl-2,FN-1,α-SMA,CollagenⅠ,IGF1,MMP1,MMP2,MMP3 and MMP7 were significantly increased(P<0.05,P<0.01).Compared with TGF-β group,the EdU positive rate and number of cell migration in OP-D group at different concentration were significantly decreased(P<0.05,P<0.01).TUNEL positive rate and Bax protein expression level were significantly increased(P<0.05,P<0.01).The protein expression levels of Bcl-2,IGF1,MMP1,MMP2,MMP3 and MMP7 were significantly decreased(P<0.05,P<0.01).The protein expression levels of FN-1 and Collagen Ⅰ in 4 μmol·L-1 OP-D group were significantly decreased(P<0.05,P<0.01),while α-SMA protein expression was significantly decreased in 2 and 4 μmol·L-1 OP-D groups(P<0.05,P<0.01).Conclusion OP-D may play an anti-IPF role by inhibiting IGF1 expression and down-regulating the level of MMPs,thereby inhibiting the proliferation,migration,apoptosis resistance and fibrosis of HFL-1 cells.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Key words
Ophiopogonin D/idiopathic pulmonary fibrosis/bioinformatics/network pharmacology/molecular docking/experimental validation/human embryonic lung fibroblasts分类
医药卫生引用本文复制引用
李仲普,黄乐,刘雨,胡学军,邓秀娟,管聘..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[J].中药新药与临床药理,2025,36(2):219-229,11.基金项目
国家自然科学基金项目(82004306) （82004306）
湖南省中医药科研计划项目(B2023017) （B2023017）
湖南省自然科学基金项目(2024JJ5237) （2024JJ5237）
