神经管缺陷胎儿孕母与正常胎儿孕母血清的差异蛋白质研究

Roles of differentially expressed proteins in maternal sera of neural tube defect fetuses

目的探讨神经管缺陷(Neural tube defects,NTDs)的产前诊断思路。方法选取2015年1月1日至2015年12月31日经产前超声诊断为NTDs胎儿的孕母血清10例作为研究组,包括脊柱裂5例,无脑儿2例,脑膨出3例。选取同期正常胎儿的孕母血清10例作为对照组。所有20例孕母均于清晨空腹取外周静脉血5 ml,离心后取血清,进行蛋白质提区,利用非标记定量技术鉴定蛋白质肽段,再结合Uniprot数据库对差异蛋白质进行生物信息学分析。结果共筛选出17个差异蛋白质,其中12个表达上调,包括妊娠带蛋白、补体C1q子组件A、B、C单元、互补DNA FLJ51564、FLJ56954、FLJ78071,血红蛋白α、β亚基,碳酸酐酶1,角蛋白Ⅱ型细胞骨架80,过氧化物氧化还原酶-2;5个表达下调,包括人冠毛素-1,S100A9蛋白,转铁蛋白,互补DNA FLJ61165、FLJ53691。差异表达蛋白质涉及生物调节、信号转导、细胞过程、细胞生长、细胞粘附、细胞组成、运输运动、催化作用和代谢等方面。结论通过非标记定量技术检测出NTDs胎儿和正常胎儿的孕母血清中存在差异表达蛋白质。

Objective To determine whether the profiling of maternal sera proteins in neural tube defects (NTDs) pregnancy is different from that of normal pregnancy controls using label-free quantitation. Methods Ten pregnant women whose fetuses were diagnosed ultrasonically with NTDs and consisting of spina bifida (n=5), anencephaly (n=2) and encephalocele (n=3) were selected along with another 10 pregnant women with normal fetuses.Samples of 5 ml peripheral venous blood of all pregnant women were harvested early in the morning.The sera samples were collected and purified by centrifugation.Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the proteins were extracted, digested by filter-aided sample prep (FASP) and followed by extraction of peptides and desalination.The proteins were then identified by label-free quantitative analysis plus LC/MS/MS-based proteomics.The differentially expressed proteins involved in biological functions were analyzed using protein bioinformatics analysis plus Unipart Database, including Gene Ontology, signaling pathways and protein interactions. Results Significant inter-group differences existed in 17 protein peaks.Among 17 proteins, 12 were up-regulated proteins/peptides, including pregnancy zone protein, complement C1q subcomponent subunit A, B & C, human cDNA FLJ51564, FLJ56954 & FLJ78071, hemoglobin subunit alpha & beta, carbonic anhydrase 1, keratin, type II cytoskeletal 80 and peroxiredoxin-2;5 were down-regulated proteins/peptides, including pappalysin-1, protein S100A9, serotransferrin, human cDNA FLJ61165 and FLJ53691.The above-mentioned proteins were involved in biological regulation, signal transduction, cellular processes, cell growth, cell adhesion, cellular components, transporter activities, catalysis activities and metabolic processes. Conclusions There are differential expression profiles of serum proteins between neural tube defect group and control group using label-free quantitation.