基于iTRAQ技术研究特发性羊水过多患者羊水蛋白组及差异蛋白的研究

Application of iTRAQ proteomics in the screening of differential proteins in amniotic fluid in patients with idiopathic polyhydramnios

目的:鉴定特发性羊水过多患者和正常妊娠孕妇羊水蛋白组学,筛选在特发性羊水过多发生发展中发挥重要作用的调控蛋白,以便深入了解羊水量调控机制。方法:收集特发性羊水过多者和正常妊娠者的羊水(6例)胎盘组织(40例),应用同位素标记相对与绝对定量(iTRAQ)技术鉴定羊水蛋白组。Western blot法检测羊膜、绒毛膜、胎盘绒毛组织中差异蛋白的表达。结果:羊水中共鉴定到1246个蛋白,357个差异蛋白,其中282个上调蛋白,75个下调蛋白(>1.2 or<0.83,P<0.05)。Western blot法结果显示,EPHB4蛋白在特发性羊水过多患者的羊膜、绒毛膜组织中表达上调,与羊水中表达水平一致。结论:首次利用iTRAQ技术鉴定特发性羊水过多患者羊水蛋白组,为羊水量的调节机制提供研究基础。EPHB4可能与羊水量异常相关。

Objective:To establish a comparative proteome profile of the amniotic fluid of idiopathic polyhydramnios and normal pregnancies and identify the responsible mediators that regulate amniotic fluid volume.Methods:A total of 6 amniotic fluids and 40 placenta samples were collected from Idiopathic Polyhydramnios and normal pregnancies.Used isobaric tag for relative and absolute quantitation(iTRAQ)proteomics technique and bioinformatics analysis to examine the proteomics in amniotic fluid.Western blot was used to verify differentially expression proteins in amnion,chorion,placenta samples.Results:We had identified 1246 proteins,357 differentially expression proteins,282 of which were upregulated and 75 downregulated(>1.2 or<0.83,P<0.05).In the verify procedure,EPHB4 protein was detected highly significant expression in amnion and chorion in IPH(P<0.05),which had some expressions trend compared with the iTRAQ results.Conclusion:We firstly employed proteome profile of the amniotic fluid of Idiopathic Polyhydramnios,and the results offer fundamental points to exploring regulate amniotic fluid volume in the future studies,EPHB4 protein may relate the disordered of amniotic volume homeostasis.