利用同位素标记相对和绝对定量技术筛查神经管畸形胚胎孕鼠血清差异蛋白质谱

Identification of Differentially Expressed Proteins in Sera of Pregnant Rats with Fetuses Having Neural Tube Defects Using Isobaric Tag Technology for Relative and Absolute Quantitation

目的筛查神经管畸形相关血清标记物,为阐明神经管畸形的发病机制提供理论依据。方法应用维甲酸在大鼠神经管闭合的关键期(孕10 d,E10)给药,建立脊柱裂大鼠模型,选取致畸早期孕11、13 d (E11、E13)显性脊柱裂胚胎孕鼠和正常胚胎孕鼠血清各5例,应用同位素标记相对和绝对定量技术(iTRAQ)和液相色谱质谱鉴定技术分析2组蛋白质,定量信息以蛋白的丰度比差异倍数1.5倍以上(P <0.05)为差异蛋白质。结果质谱共得到谱图157 422张,鉴定出390种蛋白质,发现E11维甲酸致畸组与正常对照组差异蛋白40种,E13维甲酸致畸组与正常对照组差异蛋白26种。结论筛选出致畸早期大量神经管畸形和正常孕鼠血清差异蛋白,差异蛋白通过直接或间接相互作用,促进或抑制神经管畸形的发生与发展。

Objective To screen for a potential biomarker of neural tube defects and understand the pathogenesis of this defect.Methods The rat model of spina bifida aperta(SBA)was induced with all-trans retinoic acid at E10.Serum samples were collected from normal pregnant rats and those harboring SBA fetuses at E11 and E13(n=5 each).The isobaric tag for relative and absolute quantitation(iTRAQ)proteomic approach was used to identify and quantify the differentially expressed proteins in the two groups.The quantitative protein ratios with P<0.05 and fold-changes>1.5 were considered significant in Mascot.Functional annotations of proteins were conducted.Results A total of 390 proteins corresponding to 157422 tandem mass spectrometry spectra were identified.A total of 40 proteins at E11 and 26 proteins at E13 had significant differential expression between pregnant rats carrying SBA fetuses and normal controls.Conclusion Differential serum proteome profiles associated with neural tube defects at the early stage of malformation can be identified using the iTRAQ proteomic approach,and various proteins might be involved in abnormal neurological development.