GEO芯片联合网络药理学探究六味地黄丸治疗系统性红斑狼疮作用机制及核心靶点分析

Mechanism and Core Target Analysis of Liuwei Dihuang Pills in Treatment of Systemic Lupus Erythematosus Based on GEO Chip and Network Pharmacology

目的运用网络药理学及生物信息学探究六味地黄丸治疗系统性红斑狼疮(SLE)的作用机制,并对核心靶点基因进行深入分析。方法利用TCMSP、SwissTargetPrediction、本草组鉴(HERB)数据库检索六味地黄丸的主要活性成分及其相关靶点,通过R软件limma包对GEO数据库SLE数据集GSE49454进行差异分析。利用R软件VennDiagram包进行交集靶点分析,clusterProfiler包对交集靶点基因进行GO和KEGG通路富集分析。采用Cytoscape3.8.2软件进行活性成分-靶点-通路网络构建与分析。通过STRING平台构建蛋白相互作用网络并利用Cytoscape3.8.2软件筛选核心靶点基因。利用PyMOL软件对核心活性成分与核心靶点基因进行分子对接验证;通过临床数据与TRRUST数据库进行表达量、转录因子与免疫浸润分析。结果共筛选出有效活性成分69个,相关靶点813个,基因芯片差异基因703个。KEGG通路富集分析显示,主要涉及NOD样受体等信号通路。筛选出5个核心成分(槲皮素、山柰酚、海风藤酮、玉兰脂素B、羟基芫花素)及5个核心靶点基因(TLR4、CCL2、PTPRC、CXCL10、MPO);分子对接显示,核心成分与核心靶点之间均具有稳定的结合能力。探寻到3个相关转录因子(SPI1、STAT1、NFKB1);免疫浸润分析表明,核心靶点基因与T细胞、中性粒细胞和NK细胞等关系密切。结论六味地黄丸可能从抗炎、调节免疫细胞功能等方面发挥对SLE的治疗作用。

Objective To investigate the potential mechanism of Liuwei Dihuang Pills in the treatment of systemic lupus erythematosus(SLE)through network pharmacology and bioinformatics;To conduct in-depth analysis of the core target genes.Methods The active components of Liuwei Dihuang Pills and their associated targets were retrieved through the TCMSP,SwissTargetPrediction and HERB databases.The R software VennDiagram package was used to perform intersection target analysis,and the clusterProfiler package performs GO and KEGG pathway enrichment analysis on intersecting target genes.Cytoscape 3.8.2 software was used to construct and analyze the active components-target-pathway network.Protein-protein interaction network was constructed through STRING platform,and the core target genes were screened by Cytoscape 3.8.2 software.Molecular docking verification of core active ingredients and core target genes was conducted using PyMOL software;expression levels,transcription factors and immune infiltration were analyzed from clinical data and the TRRUST database.Results A total of 69 effective active components,813 related targets,and 703 gene chip differential genes were screened out.KEGG pathway analysis showed that signaling pathways such as NOD-like receptors were mainly involved.Five core components(quercetin,kaempferol,hayfeverone,yuccinin B and hydroxyturianin)and five core target genes(TLR4,CCL2,PTPRC,CXCL10,MPO)were subsequently screened.Molecular docking showed that all core components have stable binding ability to core targets.Three related transcription factors were probed:SPI1,STAT1 and NFKB1.Immune infiltration indicated that core target genes were closely related to T cells,neutrophils and NK cells,etc.Conclusion Liuwei Dihuang Pills may exert its therapeutic effect on SLE from many aspects,such as anti inflammatory and regulating immune cell function.