摘要
目的运用网络药理学和体外实验系统探讨老鹳草治疗溃疡性结肠炎的作用机制。方法在人类基因综合分析数据库和在线人类孟德尔遗传数据库中检索和筛选了老鹳草的活性成分和相应的溃疡性结肠炎相关靶点,取交集得到重叠靶点后进一步构建基因的蛋白质互作网络,通过拓扑分析筛选出核心靶点。采用R语言进行基因本体和京都基因与基因组百科全书富集物分析。采用佛波酯将体外培养的人髓系白血病单核细胞系THP-1细胞诱导分化为M0巨噬细胞,分别加入不同浓度的老鹳草水煎液预处理,最后加入脂多糖进一步诱导为M1巨噬细胞,流式细胞术检测老鹳草水煎液对M1巨噬细胞极化的影响,酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)及实时逆转录聚合酶链反应(real-time polymerase chain reaction,qRT-PCR)检测细胞上清液中IL-1β、IL-6和IL-8的含量。结果老鹳草-溃疡性结肠炎靶点网络有7个活性成分和10个核心靶点,IL-6、IL-8和IL-1β是最重要的核心靶点。富集分析表明,IL-6、IL-1β、IL-8富集在肿瘤坏死因子信号通路。体外实验结果表明,老鹳草能显著下调IL-6、IL-8和IL-1β的表达,差异有统计学意义(P<0.01);流式细胞术结果表明老鹳草能显著抑制M1巨噬细胞极化,差异有统计学意义(P<0.01)。结论本研究提示老鹳草通过抑制M1巨噬细胞极化及通过肿瘤坏死信号通路下调促炎症因子的释放来改善溃疡性结肠炎的炎症反应。
Objective To explore the therapeutic role of Geranium wilfordii and its potential mechanism in the treatment of ulcerative colitis based on network pharmacology and in vivo experiments.Methods Chemical constituents of Geranium wilfordii were searched from TCMSP data,the gene names of target sites were extracted from UniProt database,and the disease targets of ulcerative colitis were obtained from the Online Mendelian Inheritance in Man(OMIM)database and the human diseases database.And then the overlapping genes between Geranium wilfordii potential targets and ulcerative colitis targets were used to build up the protein-protein interaction(PPI)network for analyzing their interactions and finding out the big hub genes in this network.Finally,Gene Ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses of overlapping gene targets were performed in the STRING database and visualized in R software.Subsequent in vitro experimentation was carried out in THP-1 macrophages induced with bacterial lipopolysaccharides.THP-1 macrophages were indnced to differentiate into M0 macrophages,and then treated with different concentrations of Geranium wilfordii,and finally cultured with bacterial lipopolysaccharides to induce M1 macrophages.Then flow cytometric analysis was conducted to detect M1 macrophages,and enzyme-linked immunosorbent assay(ELISA)and quantitative real time-polymerase chain reaction(qRT-PCR)were used to detect the levels of IL-6,IL-8 and IL-1β.Results According to the topological parameters of PPI network and the compound-target-pathway network,10 core targets and 7 bioactive compounds were identified.The enrichment analysis of KEGG pathways demonstrated that these core targets were mainly enriched in the tumor necrosis factor(TNF)signaling pathway.In vitro,Geranium wilfordii downregulated the expression and the secretion of IL-6,IL-8 and IL-1β(P<0.01).The flow cytometry results indicated that Geranium wilfordii significantly inhibited macrophage polarization(P<0.01).Conclusion
作者
付冲
吴超
张焰平
FU Chong;WU Chao;ZHANG Yanping(Department of Gastroenterology,Anqing Municipal Hospital,Anqing 246000,China;Department of Pathophysiology,Wannan Medical College)
出处
《山西医科大学学报》
CAS
2022年第10期1276-1285,共10页
Journal of Shanxi Medical University
基金
安徽省重点研究与开发计划项目(S202104j07020117)
安庆市立医院院级科研项目(2022aqykj19)。
关键词
网络药理学
体外实验
老鹳草
溃疡性结肠炎
network pharmacology
vitro experiment
Geranium wilfordii
ulcerative colitis
