基于iTRAQ技术对戊四氮点燃癫痫大鼠海马组织的蛋白质组学分析  

iTRAQ-based proteomic analysis of hippocampus ofpentylenetetrazol-kindled epileptic rats

作　　者：张鹏 张丽 熊红丽 朱士胜[4] ZHANG Peng;ZHANG Li;XIONG Hongli;ZHU Shisheng(Department of Forensic Medicine,Hainan Medical University,Haikou 571199;School of Basic Medicine,Chongqing College of Traditional Chinese Medicine,Chongqing 402760;Department of Forensic Medicine,Faculty of Basic Medical Sciences,Chongqing Medical University,Chongqing 400016;College of Basic Medical Sciences,Chongqing Medical University,Chongqing 401331,China)

机构地区：[1]海南医学院法医学教研室,海口571199 [2]重庆中医药学院基础医学部,重庆402760 [3]重庆医科大学基础医学院,重庆400016 [4]重庆医药高等专科学校基础部,重庆401331

出　　处：《神经解剖学杂志》2023年第6期649-656,共8页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81901922);海南省高等学校科学研究重点资助项目(Hnky2020ZD-18);重庆市沙坪坝区决策咨询与管理创新项目(Jcd202042);重庆医药高等专科学校创新研究群体项目(YGZ2019402);重庆医药高等专科学校科研计划项目(ygz2018103,ygz2020102,ygz2021119)。

摘　　要：目的:筛选癫痫大鼠海马组织差异表达蛋白(DEPs),为进一步探索癫痫的发病机制和药物治疗靶点提供思路。方法:采用戊四氮(PTZ)诱导建立SD大鼠癫痫模型(PTZ组),利用同位素标记相对和绝对定量(iTRAQ)联合LC-MS/MS技术检测大鼠海马组织蛋白质谱,以PTZ组对control组蛋白表达量变化倍数>1.5或<0.67且P<0.05为标准筛选DEPs,并对DEPs进行基因本体(GO)功能注释和京都基因与基因百科全书(KEGG)通路富集等生物信息学分析。结果:共筛选出80个显著DEPs,其中上调39个,下调41个。GO分析显示:上调的DEPs主要涉及细胞对神经生长因子刺激的反应、轴突发育、细胞表面受体信号通路、神经元迁移、肌动蛋白细丝解聚和信号转导等生物过程;下调的DEPs主要涉及三羧酸循环、柠檬酸盐代谢过程、丙酮酸-乙酰辅酶A生物合成过程、草酰乙酸代谢过程、粘附聚集体的调节等生物过程。KEGG通路富集分析显示:上调的DEPs主要参与腺苷酸活化蛋白激酶(AMPK)信号通路、肌动蛋白细胞骨架的调节、鞘脂信号通路、苯丙氨酸代谢和胰岛素信号通路等5条信号通路;下调的DEPs主要参与柠檬酸循环、碳代谢、乙醛酸和二羧酸代谢、代谢途径、氨基酸的生物合成和肌萎缩侧索硬化症等6条信号通路。结论:本研究通过iTRAQ蛋白质组学方法筛选出了癫痫海马组织的DEPs,对DEPs进行生物信息学分析所富集的代谢通路可能与癫痫的发病密切相关。Objective:To screen differentially expressed proteins(DEPs)in the hippocampus of epileptic rats,providing ideas for further exploring the pathogenesis and drug therapeutic targets of epilepsy.Methods:The epileptic rat model(PTZ group)induced by pentylenetetrazol(PTZ).The protein profiles of hippocampal tissues of the PTZ group and control groups were detected by isobaric tag for relative and absolute quantitation(iTRAQ)combined with LC-MS/MS technology,and the fold changes in the protein expression of the PTZ group versus the control group was considered to be more than 1.5 or less than 0.67,and P<0.05 was taken as the criteria for screening DEPs.Then DEPs were subjected to bioinformatics analyses such as Gene Ontology(GO)functional annotation and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.Results:A total of 80 proteins showed significantly different expression,which included 39 up-regulated and 41 down-regulated.GO analysis showed that the up-regulated DEPs were mainly involved in biological processes such as cellular response to nerve growth factor stimulus,axon development,cell surface receptor signaling pathway,neuron migration,actin filament depolymerization,and signal transduction,while down-regulated DEPs were mainly involved in biological processes such as tricarboxylic acid cycle,citrate metabolism process,acetyl-CoA biosynthetic process from pyruvate,oxaloacetate metabolism process,and regulation of focal adhesion assembly.KEGG pathway enrichment analysis showed that up-regulated DEPs were mainly involved in five signaling pathways,including AMPK signaling pathway,regulation of actin cytoskeleton,sphingolipid signaling pathway,phenylalanine metabolism,and insulin signaling pathway;The down-regulated DEPs were mainly involved in six signaling pathways,including the citric acid cycle,carbon metabolism,acetate and dicarboxylate metabolism,metabolic pathways,amino acid biosynthesis and amyotrophic lateral sclerosis signaling pathways.Conclusion:DEPs from epileptic hippocampal t

关 键 词：癫痫 蛋白质组学 同位素标记相对和绝对定量 生物信息学分析 戊四氮点燃模型 大鼠 

分 类 号：R742.1[医药卫生—神经病学与精神病学]