摘要
目的:基于生物信息学和体外实验探究粉防己碱治疗早期矽肺发展的关键通路。方法:通过文献挖掘收集矽肺患者的差异表达基因;利用高通量基因表达数据库(GEO)收集二氧化硅滴注小鼠的差异表达基因;借助在线人类孟德尔遗传数据库(OMIM)、GeneCards、比较毒物基因组学数据库(CTD)获取矽肺相关的疾病靶点;分别将差异表达基因和疾病靶点通过R语言及Metascape平台进行基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)富集分析;使用薛定谔(Schrödinger)及Pymol软件进行分子对接及修饰;制备二氧化硅刺激的巨噬细胞和上皮细胞模型,通过PCR技术和蛋白免疫印迹(WB)验证分析结果。结果:筛选出矽肺病人差异表达基因2065个,滴注二氧化硅大鼠差异表达基因2291个,矽肺相关疾病靶点803个。GO富集分析主要涉及G蛋白偶联受体结合、调节炎症反应、参与免疫反应等。KEGG通路富集分析主要包括细胞外基质-受体相互作用(ECM-receptor interaction)、TNF信号通路、IL-17信号通路等。不同来源的差异基因和疾病靶点同时筛选出IL-17信号通路。分子对接结果表明矽肺药物粉防己碱与IL-17信号通路中的RAF/MEK/ERK通路有良好的结合效果。细胞实验表明粉防己碱通过调控RAF/MEK/ERK通路降低巨噬细胞中TNF-α、TGF-β等炎症因子的表达,同时抑制上皮细胞中上皮间质转化及炎症因子表达。结论:粉防己碱通过RAF/MEK/ERK通路调控炎症反应和上皮间质转化(EMT)从而影响早期矽肺进展。
Objective:Exploring the key pathways affecting the development of early silicosis based on bioinformatics and in vitro experiments.Method:Collecting differentially expressed genes in silicosis patients through literature mining;Collecting differentially expressed genes in silicon dioxide infusion mice by using a high-throughput gene expression database(GEO);Obtaining disease targets related to silicosis by means of online human Mendelian genetic database(OMIM),GeneCards and comparative toxicgenomics database(CTD);differentially expressed genes and disease targets were subjected to gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genome(KEGG)enrichment analysis via R-package and Metascape platforms,respectively.The Schrödinger and Pymol software were used for molecular docking and modification.Silicon dioxide-stimulated macrophages and epithelial cells were modeled and analyzed by PCR and western blot(WB).Result:2065 differentially expressed genes in silicosis patients,2291 differentially expressed genes in rat infused with silicon dioxide,and 803 targets for silicosis-related diseases were screened out.GO enrichment analysis mainly involves G protein-coupled receptor binding,the regulation of inflammatory response,and participation in immune response.The enrichment analysis of KEGG pathway mainly included ECM-receptor interaction,TNF signaling pathway,and IL-17 signaling pathway.IL-17 signaling pathway was screened out from different genes and disease targets,indicating that IL-17 signaling pathway might be the key pathway for the development of silicosis.Molecular docking results showed that the silicosis drug tetrandrine had good binding effect with the RAF/MEK/ERK pathway in the IL-17 signaling pathway.Cellular experiments showed that tetrandrine reduced the expression of inflammatory factors such as IL-6 and TGF-βin macrophages by regulating the RAF/MEK/ERKpathway,and inhibited the epithelial-mesenchymal transition and expression of inflammatory factors in epithelial cells.Conclusion
作者
梁超
周家伟
刘亚锋
郭健强
王清森
苏忆欣
邢应如
胡春晓
谢军
吴静
胡东
LIANG Chao;ZHOU Jia-wei;LIU Ya-feng;GUO Jian-qiang;WANG Qing-sen;SU Yi-xin;XING Ying-ru;HU Chun-xiao;XIE Jun;WU Jing;HU Dong(School of Medicine,Anhui University of Science and Technology,Huainan 232001,China;Anhui Occupational Health and Safety Engineering Laboratory,Huainan 232001,China;Key Laboratory of Advanced Industrial Dust Purification and Occupational Health and Safety of Education Department,Huainan 232001,China;Department of Clinical Laboratory,Gengjiu Hospital of Anhui Zhongke,Hefei 232001,China;Affiliated Cancer Hospital of Anhui University of Science and Technology/Huainan Oriental Hospital Group Cancer Hospital,Huainan 232001,China)
出处
《海南医学院学报》
CAS
2022年第23期1792-1802,共11页
Journal of Hainan Medical University
基金
国家自然科学基金项目(81971483)
安徽省高校协同创新项目(GXXT-2020-058)
安徽理工大学大学生创新创业项目(2021CX2125,2021CX2126,2021CX2124)。
关键词
矽肺
粉防己碱
生物信息学
分子对接
信号通路
Silicosis
Tetrandrine
Bioinformatics
Molecular docking
signal pathway
