先天性肾盂输尿管连接部狭窄的比较蛋白质组学研究  

Comparative proteomic analysis of congenital ureteropelvic junction stenosis

作　　者：刘靖 刘明学[2] 杨振宇[2] 苏毅[2] 纪延辉[2] 覃宇冰[2] 黄文倩[2] 何朝升[2] 黄桂珍 杨国柱 胡增隆 Liu Jing;Liu Mingxue;Yang Zhenyu;Su Yi;Ji Yanhui;Qin Yubing;Huang Wenqian;He Chaosheng;Huang Guizhen;Yang Guozhu;Hu Zenglong(Department of Laboratory Medicine,The First Affiliated Hospital of Xiamen University,Xiamen 361003,China;Department of Pediatrics,The First Affiliated Hospital of Xiamen University,Xiamen 361003,China)

机构地区：[1]厦门大学附属第一医院检验科,厦门361003 [2]厦门大学附属第一医院儿外科,厦门361003

出　　处：《临床小儿外科杂志》2022年第5期474-481,共8页Journal of Clinical Pediatric Surgery

基　　金：福建省自然科学基金(2019J01564)。

摘　　要：目的对先天性肾盂输尿管连接部(ureteropelvic junction,UPJ)狭窄患儿狭窄段输尿管组织与邻近正常输尿管组织进行比较蛋白质组学研究,探索先天性UPJ狭窄的可能病因及发病机制。方法以厦门大学附属第一医院收治的30例先天性UPJ狭窄并在本院行Anderson-Hynes术的患儿为研究对象,标本离体后立即取狭窄段输尿管组织(A组,n=30)及与狭窄段远端邻近的正常输尿管组织(B组,n=30)。采用基于串联质谱标签(tandem mass tag,TMT)标记的高通量蛋白质组相对定量方法对两组中的差异蛋白进行高通量筛选,随后对筛选出的差异蛋白进行平行反应监测(parallel reaction monitoring,PRM)的靶向蛋白质组学定量验证,利用生物信息学方法对差异蛋白进行基因本体(gene ontology,GO)功能注释、京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路注释的富集分析,并对蛋白质相互作用(protein-protein interaction,PPI)网络进行分析。结果两组标本共鉴定出蛋白质6093种,包含差异蛋白160种,其中表达上调的65种,表达下调的95种。进行PRM相对定量分析的9种目标蛋白中,碳酸酐酶1等7种蛋白质在A组与B组的蛋白含量比值,在PRM和TMT的定量结果中均小于1;磷脂酰肌醇蛋白聚糖-6和具有多重剪接的RNA结合蛋白在A组与B组的蛋白含量比值,在PRM和TMT的定量结果中均大于1。GO功能富集分析发现,两组中差异表达蛋白质在体液免疫反应、抗微生物体液反应、对真菌的反应、对真菌的防御反应和对细菌的防御反应等生物学过程的GO功能富集度具有高的显著性水平(P<0.05)。KEGG通路富集分析结果表明,氮代谢、PPAR信号通路等20条信号通路富集度具有高的显著性水平(P<0.05)。在PPI网络中,连接度最高的蛋白质是:中性粒细胞弹性蛋白酶(连接度为9)和组织蛋白酶抑制素抗菌肽(连接度为8)。结论比较蛋白质组学研究结果提示,先天Objective To conduct comparative proteomics study between the stenotic and the normal ureteral tissues adjacent to the stenotic ureter in children with ureteropelvic junction(UPJ)stenosis,and explore the etiology and pathogenesis of congenital UPJ stenosis.Methods 30 children with congenital UPJ stenosis underwent Anderson-Hynes operation.The ureteral tissue of the narrow segment(group A)and the normal ureteral tissue adjacent to the distal end of the stenosis(group B)were taken immediately after the operation.A relative quantitative method of high-throughput proteomics based on tandem mass tag(TMT)was used to screen the differentially expressed proteins in these two groups.And then the targeted proteomics quantitative verification of the screened differential proteins was carried out by parallel reaction monitoring(PRM).Gene Ontology(GO)functional annotation,Kyoto Encyclopedia of genes and genomics(KEGG)pathway were enriched and analyzed by bioinformatics method,and protein-protein interaction(PPI)network was analyzed.Results A total of 6093 proteins were identified in the two groups of specimens,including 160 differential proteins,of which 65 were up-regulated and 95 were down-regulated.Among the 9 target proteins subjected to PRM relative quantitative analysis,the ratio of protein content of 7 proteins including carbonic anhydrase 1 in group A and group B was less than 1 in the quantitative results of PRM and TMT;Glypican The ratio of protein content of-6 and RNA-binding proteins with multiple splicing in group A to group B was greater than 1 in the quantitative results of PRM and TMT.GO functional enrichment analysis found that the GO functional enrichment of differentially expressed proteins in the two groups in biological processes such as humoral immune response,anti-microbial humoral response,response to fungi,defense response to fungi,and defense response to bacteria were significantly higher.High level of significance(P<0.05).The results of KEGG pathway enrichment analysis showed that the enrichment of

关 键 词：肾盂/外科学 输尿管疾病/并发症 蛋白质组学 计算生物学 

分 类 号：R726.8[医药卫生—儿科]