【摘要】 目的应用同位素标记的绝对和相对定量(isobaric tags for relative and absolute quantitation,iTRAQ)技术筛选并鉴定胸科手术中分别采用单肺通气和双肺通气的患者血清中差异表达的蛋白质。方法收集胸科手术中采用单肺通气和双肺通气的患者血清,每组各10例,将每一组血清等量混合后应用多重免疫亲和层析柱(MARS)去除血清中的高丰度蛋白,样本经iTRAQ标记和液相色谱分离后,用质谱仪进行鉴定和相对定量分析。结果经质谱鉴定共获得189种蛋白质,其中差异表达的蛋白质有42种(上调或下调20%)。单肺通气组与双肺通气组比较,有33种蛋白质表达上调,9种表达下调。结论iTRAQ技术能够快速、有效地进行差异蛋白质组学的研究,能筛选出多种与单肺通气肺损伤相关的生物标记蛋白,为进一步研究单肺通气肺损伤发生的机制提供实验依据。更多还原

【Abstract】 Objective To identify serum proteins differentially expressed in patients undergoing different ventilation modes during thoracic operations. Methods Ten patients underwent one-lung ventilation(group A) and 10 underwent two-lung ventilation(group B) during thoracic operations. Serum samples were collected, and highly abundant serum proteins were purified on an immunoaffinity chromatography column.The iTRAQ technique combined with two-dimensional liquid chromatography tandem mass spectrometry (2D LC-MS/MS) was used to identify and quantitate proteins differentially expressed between the two groups. Results A total of 189 pro-teins were identified by mass spectrometry,and 42 proteins were differentially up-or down-regulated by at least 20%.Of these proteins,33 were up-regulated and 9 were down-regulated in group A compared to group B. Conclusions The iTRAQ technique is effective for comparative proteomic studies.Protein expression profiling may prove a useful approach for investigating the pathogenesis of lung injury in patients undergoing one-lung ventilation.更多还原