复发性流产患者胎盘绒毛组织中差异表达蛋白的蛋白质组学分析

Proteomics analysis of differentially-expressed proteins in placental villous tissue from recurrent spontaneous abortion patients

关键词:  生物信息学分析  胎盘绒毛  蛋白组学  复发性流产

Key words:  bioinformatics analysis  placental villus  proteomics  recurrent spontaneous abortion

摘要:

目的通过蛋白质组学及生物信息学分析等技术手段，探索复发性流产（recurrent spontaneous abortion，RSA)的发病机制。方法对RSA组和正常妊娠组的胎盘绒毛组织进行蛋白组学分析，鉴定二者中差异表达的蛋白，同时对这些差异表达的蛋白进行基因相互作用（KEGG）富集分析、信号网络分析（ingenuity pathway analysis， IPA)、蛋白互作（protein-protein interaction， PPI）分析等，找出富集通路。通过PPI网站及cytoscape软件，从这些差异蛋白中找出了一些枢纽蛋白，将得分前20的枢纽蛋白和富集最明显KEGG富集通路进行匹配，取它们的交集，从3个通路里分别挑选出对应的3个蛋白，最后用Western印迹法对这3个蛋白进行验证。结果在两组绒毛组织中总共鉴定出了6 438种蛋白质，其中有342个在两组绒毛组织中是差异表达的，共208个上调，134个下调。关于差异表达蛋白的KEGG富集分析显示，上调的蛋白富集最明显的通路为补体和凝血系统，而下调的蛋白富集最明显的通路为核糖体通路，这3个通路对应的枢纽蛋白分别为补体C3、抗凝血酶Ⅲ（AT-Ⅲ）、线粒体核糖体蛋白S7（MRPS7）。对这3个蛋白进行的Western印迹法结果显示，MRPS7在RSA组绒毛组织中下调，补体C3、AT-Ⅲ在RSA组中上调，这一结果与蛋白组学检测的结果一致。结论在RSA胎盘绒毛组织中存在补体系统和凝血系统的上调以及核糖体通路的下调，表明补体系统、凝血系统和核糖体通路的失调可能是导致RSA的重要原因之一。

Abstract:

ObjectiveTo explore the pathogenesis of recurrent spontaneous abortion（RSA) by proteomics, bioinformatics, and other technical approaches. MethodsThe proteomic analysis was performed on the placental villus tissues from RSA patients（RSA group) and normal pregnant women（control group) and the differentially expressed proteins were identified between RSA and control groups. These differentially expressed proteins were further analyzed by bioinformatics, including Kyoto Encyclopedia of Genes and Genomes（KEGG) pathway enrichment analysis, signal network analysis（ingenuity pathway analysis, IPA), and protein-protein interaction（PPI) analysis. Then, a PPI network was constructed using Cytoscape software to identify hub proteins from these differentially expressed proteins. The highly scored 20 hub proteins were matched with the most significant enrichment pathways of KEGG. Finally, the ultimately selected three hub proteins were verified by Western blotting. ResultsA total of 6 438 proteins were identified in placental villus tissues of both groups by proteomic analysis, of which 342 proteins were differentially expressed between the two groups including 208 upregulated proteins and 134 downregulated proteins. The KEGG enrichment analysis revealed that the upregulated proteins were mostly part of the complement system and the coagulation system, while the downregulated proteins were mainly enriched in the ribosomal pathway. The complement component 3（C3), antithrombin-Ⅲ(AT-Ⅲ), mitochondrial ribosomal protein S7（MRPS7) were selected as hub proteins from the three pathways accordingly. Western blotting confirmed that MRPS7 was down-regulated in the placental villus tissues of the RSA group, while the other two proteins were up-regulated. These results were consistent with the proteomic detection. ConclusionOur study has identified the upregulation of proteins from the complement system and coagulation system, and the downregulation of proteins from the ribosomal pathway in the placental villus of the RSA patients compared with the normal pregnant women, suggesting that the dysfunction of the complement system, coagulation system, and ribosome pathway may play a critical role in RSA.