同位素标记相对和绝对定量蛋白组学技术筛选儿童哮喘不同病情控制水平血清差异蛋白

Screening Serum Differential Proteins for Childhood Asthma at Different Control Levels by Isobaric Tags for Relative and Absolute Quantification-based Proteomic Technology

目的从整体蛋白水平研究不同控制水平儿童哮喘患者血清的差异性蛋白,为防治儿童哮喘提供依据。方法采用同位素标记相对和绝对定量标记结合二维液相色谱串联质谱定量蛋白组学技术筛选鉴定控制、部分控制和未控制儿童哮喘患者血清中差异表达蛋白,通过酶联免疫吸附测定法进行差异蛋白验证。结果共鉴定蛋白260种,筛选出不同控制水平儿童哮喘间两两比较均有差异的蛋白57种(变化倍数<0.8或变化倍数>1.2),主要参与21种生物过程,主要具有8种分子功能类型,主要为17种细胞成分类型。酶联免疫吸附测定结果显示控制组的玻连蛋白含量(573.92±412.43)μg/ml高于未控制组的(382.27±238.64)μg/ml,差异有统计学意义(P=0.0399)。结论发现不同病情控制水平儿童哮喘患者血清中的差异性蛋白57种,这些差异蛋白可作为控制儿童哮喘的潜在生物学靶点。

Objective To screen serum differential proteins for childhood asthma at different control levels,which provided the basis for the prevention and treatment of childhood asthma.Methods Isobaric tags for relative and absolute quantification,two-dimensional liquid chromatography,nanoelectrospray ionization,and high-resolution tandem mass spectrometry using the hybrid quadrupole time-of-flight platform was used to screen the differential proteins in serum samples from pediatric patients with controlled,partly controlled,or uncontrolled childhood asthma.Differential proteins were validated using enzyme-linked immunosorbent assay（ELISA）.Results A total of 260 expressed proteins were identified.Among them 57 differentially expressed proteins were found among the different control levels of childhood asthma（fold0.8 or fold1.2）.The differentially expressed proteins were involved mainly in 21 biological processes and 8 molecular functions and were located in 17 cellular components.ELISA showed that the serum vitronectin level was significantly higher in controlled group [（573.92±412.43） μg/ml] than in uncontrolled group [（382.27±238.64）] μg/ml（P=0.0399）.Conclusion We identified 57 differential proteins for childhood asthma at different control levels,which may be used as potential biological targets for the control of childhood asthma.