基于4D-Label-free技术慢性弥漫性轴索损伤大鼠海马组织的蛋白质组学分析

Proteomic analysis of hippocampal tissue in rats with chronic diffuse axonal injury based on 4D-Label-free technique

目的筛选慢性弥漫性轴索损伤(DAI)大鼠海马组织差异表达蛋白(DEPs),为探索慢性DAI潜在发病机制以及临床诊断、筛选药物治疗靶点、评估预后等提供实验依据。方法实验动物分为模型组(DAI组,n=20)和对照组(CON组,n=20),采用改良的Marmarou法建立SD大鼠DAI模型,模型建立3周后利用4D-Label-free技术检测慢性DAI组脑海马组织中的蛋白图谱变化,以DAI组/CON组表达量变化倍数(FC)>1.2或<0.83且P<0.05筛选DEPs,运用基因本体(GO)功能注释和京都基因与基因百科全书(KEGG)通路富集分析方法对筛选的DEPs进行生物信息学分析。结果共筛选出92个DEPs,上调52个,下调40个。GO分析结果显示,DEPs主要涉及去磷酸化,ATP合成耦合电子传递,过氧化氢介导的细胞程序性死亡的正向调节及神经递质受体内化等生物过程功能。KEGG通路分析结果提示,DEPs主要参与代谢途径、活性氧、神经变性途径-多种疾病、逆行内源性大麻素信号传导、谷胱甘肽代谢等信号通路。结论通过4D-Label-free技术筛选出了慢性DAI组大鼠海马组织中的DEPs。所筛选出的DEPs及其所富集的生物过程和信号通路为慢性DAI的深入研究提供依据。

Objective To screen differentially expressed proteins(DEPs)in the hippocampal tissues of rats with chronic diffuse axonal injury(DAI),in order to provide an important theoretical basis for exploring the potential pathogenesis of chronic DAI as well as for clinical diagnosis,screening of drug therapeutic targets,and assessment of prognosis.Methods The experimental animals were divided into model group(DAI group,n=20)and control group(CON group,n=20).A modified Marmarou method was used to establish a DAI model in SD rats.After three weeks of modeling,the changes in protein profiles of hippocampal tissues between two groups were compared with the 4D-Label-free technology,and DEPs were screened by the fold change(FC)in expression of the DAI group/CON group of>1.2 or<0.83 with P<0.05.Bioinformatical analysis of selected DEPs was performed by using gene ontology(CO)functional annotation and Kyoto Encyclopedia of Genes and Genes(KEGG)pathway enrichment analysis method.Results A total of 92 DEPs were screened with 52 up-regulated and 40 down-regulated.The result of GO analysis showed that DEPs were mainly involved in biological process(BP)functions such as dephosphorylation,ATP synthesis-coupled electron transfer,positive regulation of hydrogen peroxide-mediated programmed cell death,and internalization of neurotransmitter receptors.The results of KEGG pathway analysis suggested that DEPs were mainly involved in signaling pathways such as metabolic pathways,reactive oxygen species,neurodegenerative pathways-multiple diseases,intraretrograde cannabinoid signaling,and glutathione metabolism.Conclusion The DEPs in hippocampus of chronic DAI group rats are screened by 4D-Labelfree technique.The selected DEPs and their enriched BP and signaling might pathway provide theoretical basis for further study of chronic DAI.