肝缺血再灌注致大鼠海马损伤的差异表达蛋白  

Identification of differentially expressed proteins in hippocampal injury induced by liver ischemia-reperfusion in rats

作　　者：韩文惠 贾莉莉[2] 付钰棠 刘俊鹏 孙英 盛明薇 吕丹[2] 张涛[1] 喻文立 Han Wenhui;Jia Lili;Fu Yutang;Liu Junpeng;Sun Ying;Sheng Mingwei;Lyu Dan;Zhang Tao;Yu Wenli(Nankai University School of Medicine,Tianjin 300071,China;Department of Anesthesiology,Tianjin First Central Hospital,Nankai University,Tianjin 300192,China)

机构地区：[1]南开大学医学院,天津300071 [2]南开大学附属天津市第一中心医院麻醉科,天津300192

出　　处：《中华麻醉学杂志》2024年第11期1369-1374,共6页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金面上项目(82072219);天津市自然科学基金面上项目(21JCYBJC00150);天津市卫生健康科技项目(TJWJ2023QN025)。

摘　　要：目的筛选肝缺血再灌注致大鼠海马损伤的差异表达蛋白。方法清洁级健康幼年雄性SD大鼠18只,2周龄,体质量20~30 g,采用随机数字表法分为2组(n=9):假手术组(S组)和肝缺血再灌注组(IR组)。采用肝缺血1 h恢复灌注的方法制备大鼠肝缺血再灌注损伤模型。于再灌注3 d时麻醉大鼠处死后分离海马组织,分析获得基因表达谱,R软件筛选差异表达基因,进一步通过Cytoscape和京都基因与基因组百科全书(KEGG)通路分析,构建蛋白质相互作用网络,确定差异表达显著蛋白,采用qPT-PCR和Western blot法验证。结果海马组织蛋白质组学分析共筛选出45个差异表达蛋白,包括36个表达显著上调的蛋白及9个表达显著下调的蛋白,通过CytoHubBA插件cystscape从PPI网络复合体中筛选出和损伤相关表达显著的蛋白:Ras相关C3肉毒菌毒物底物(RAC2)、HRAS、磷脂酰肌醇-3-激酶抑制蛋白(PTEN)和N-甲基-D-天氡氨酸离子能谷氨酸受体2b(GRIN2b)。qPT-PCR和Western blot结果表明,与S组相比,IR组大鼠海马组织RAC2、HRAS、PTEN和GRIN2b及其mRNA表达上调(P<0.05)。KEGG通路分析结果表明,差异表达蛋白在癌症中PD-L1的表达及检查点通路、长时程增强作用、Wnt信号通路的调节中显著富集。结论肝缺血再灌注致大鼠海马损伤的机制可能与RAC2、HRAS、PTEN和GRIN2b表达上调有关。Objective To identify the differentially expressed proteins that caused hippocampal damage after liver ischemia-reperfusion(I/R)in rats.Methods Eighteen clean-grade healthy juvenile male Sprague-Dawley rats,aged 2 weeks,weighing 20-30 g,were divided into 2 groups(n=9 each)using a random number table method:sham operation group(S group)and liver I/R group(IR group).A rat model of liver I/R injury was prepared by restoring perfusion after 1 h of liver ischemia.The rats were sacrificed after being anesthetized at day 3 of reperfusion,and the hippocampal tissue was isolated and analyzed to obtain gene expression profiles.Differentially expressed genes were identified using the R software,and further protein interaction networks were constructed through Cytoscape and Kyoto Encyclopedia Genes and Genomes pathway analysis to determine the differentially expressed proteins.Quantitative real-time polymerase chain reaction and Western blot were used for validation.Results A total of 45 differentially expressed proteins were identified by the proteomic analysis of hippocampal tissues,including 36 significantly up-regulated proteins and 9 significantly down-regulated proteins.The proteins with significant expression related to injury were identified from the PPI network complex using the CytoHubBA plug-in cystscape:Ras-related C3 botulinum toxin substrate(RAC2),HRAS,phosphatidylinositol-3-kinase inhibitor phosphatase and tensin homologue(PTEN),and N-methyl-D-aspartate ionotropic glutamate receptor 2b(GRIN2b).The results of quantitative real-time polymerase chain reaction and Western blot showed that the expression of RAC2,HRAS,PTEN,and GRIN2b in the hippocampal tissue was significantly up-regulated in IR group compared with S group(P<0.05).The results of Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that differentially expressed proteins were significantly enriched in the expression of PD-L1 and its checkpoint pathway,long-term potentiation,and regulation of the Wnt signaling pathway in cancer.Conclusions T

关 键 词：肝 再灌注损伤 海马 生物信息学分析 

分 类 号：R575[医药卫生—消化系统]