盐酸丁卡因诱导的巨噬细胞转录组变化及可变剪接事件分析  

Analysis of Transcriptome Changes and Alternative Splicing Events Induced by Tetracaine Hydrochloride in Macrophages

作　　者：秦正山 张燃 王唯一 赵鑫 易鹏 冯建国 QIN Zhengshan;ZHANG Ran;WANG Weiyi;ZHAO Xin;YI Peng;FENG Jianguo(Department of Anesthesiology,The Affiliated Hospital,Southwest Medical University,Luzhou 646000,China;Anesthesiology and Critical Care Medicine Key Laboratory of Luzhou,The Affiliated Hospital,Southwest Medical University,Luzhou 646000,China;Department of Anesthesiology,Suining Central Hospital,Suining 629099,China)

机构地区：[1]西南医科大学附属医院麻醉科,泸州646000 [2]西南医科大学附属医院麻醉与重症医学泸州市重点实验室,泸州646000 [3]遂宁市中心医院麻醉科,遂宁629099

出　　处：《西南医科大学学报》2025年第2期177-183,共7页Journal of Southwest Medical University

基　　金：泸州市人民政府-西南医科大学科技战略合作项目(2021LZXNYD-D08);四川省大学生创新创业训练计划项目(S202310632238)。

摘　　要：目的探究盐酸丁卡因对巨噬细胞基因表达谱及其对RNA可变剪接(alternative splicing,AS)的影响。方法采用转录组测序技术对盐酸丁卡因(200μM)处理24 h的小鼠巨噬细胞(RAW 264.7)进行分析。筛选差异表达变化超过2倍的m RNA,并通过生物信息学工具进行基因本体(gene ontology,GO)功能分类、京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集、反应途径分析及差异AS事件分析。结果经丁卡因处理RAW 264.7后,共识别出差异表达基因52119条,包括1785条上调基因和1454条下调基因。GO分析显示主要富集在DNA复制中的DNA解旋、T细胞趋化、DNA复制起始和DNA双链解旋等生物过程。KEGG通路分析揭示主要富集通路包括核糖体、新冠肺炎、氧化磷酸化、细胞周期、p53信号通路和动物线粒体自噬等。Reactome富集到无义介导的mRNA降解、信号识别颗粒(signal recognition particle,SRP)依赖的共翻译蛋白质靶向膜、rRNA加工等途径。基因集富集分析(gene set enrichment analysis,GSEA)显示免疫炎症反应和p53通路等被激活,而细胞周期通路被抑制。差异AS分析指出丁卡因处理显著影响巨噬细胞的AS事件。结论局麻药丁卡因显著影响了巨噬细胞的基因表达谱,差异基因与DNA损伤修复、细胞周期、免疫应答、r RNA加工、RNA无义降解等密切相关,同时丁卡因也显著影响了巨噬细胞中的基因AS过程。Objective To investigate the effects of tetracaine hydrochloride on gene expression profiles and RNA alternative splicing patterns in macrophages.Methods In this study,we utilized transcriptome sequencing technology to analyze mouse macro⁃phage RAW 264.7 cells treated with 200μM tetracaine hydrochloride for 24 hours.Differentially expressed mRNAs showing changes greater than two-fold were identified and analyzed using bioinformatics tools for GO functional categorization,KEGG pathway enrich⁃ment,Reactome pathway analysis,and differential alternative splicing events analysis.Results After tetracaine hydrochloride treat⁃ment of RAW 264.7 cells,a total of 52119 differentially expressed genes were identified,including 1785 upregulated and 1454 down⁃regulated genes.GO analysis indicated significant enrichment in biological processes involved in DNA unwinding,DNA replication,T cell chemotaxis,DNA replication initiation,and DNA duplex unwinding.KEGG pathway analysis revealed enrichment in pathways such as Ribosome,Coronavirus disease-COVID-19,Oxidative phosphorylation,Cell cycle,p53 signaling pathway,and animal Mitophagy.Reactome analysis showed enrichment in processes such as Nonsense Mediated Decay(NMD)independent of the Exon Junction Complex(EJC),SRP-dependent co-translational protein targeting to membrane,and rRNA processing.GSEA analysis indi⁃cated activation of immune-inflammatory responses and the p53 pathway,while the cell cycle pathway was suppressed.Differential alternative splicing analysis indicated that tetracaine treatment significantly affected alternative splicing events in macrophages.Con⁃clusion The local anesthetic tetracaine hydrochloride significantly affected the gene expression profile of macrophages.The differen⁃tially expressed genes were closely related to DNA damage repair,cell cycle,immune response,ribosomal RNA processing,and nonsense-mediated RNA decay.Additionally,tetracaine also significantly influenced the alternative splicing process in macrophages.

关 键 词：局麻药 盐酸丁卡因 巨噬细胞 转录组 差异基因 

分 类 号：R614.3[医药卫生—麻醉学]