应用TMT结合质谱技术筛选高原红细胞增多症患者血浆差异表达蛋白

摘要

AIM:To screen differentially expressed proteins in plasma of the patients with high-altitude polycythemia (HAPC) and healthy people by the technique of tandem mass tags (TMT) combined with mass spectrometry.METHODS:Four HAPC patients and five matched healthy individuals were enrolled, and their plasma samples were obtained.After balanced mixing, multiple immune affinity chromatography column was used to remove 14 kinds of high-abundant proteins, and HPLC-MS/MS using an isobaric TMT proteomic technology was performed to indentify the differentially expressed proteins among groups.The concentrations of C-reactive protein (CRP) and serum amyloid A (SAA) proteins in the plasma in 20 HAPC patients and 20 health controls were measured by ELISA.RESULTS:A total of 1 094 kinds of proteins were identified, and 249 differentially expressed proteins were screened, with 162 kinds of proteins up-regulated by ≥1.5 folds and 87 kinds of proteins down-regulated by ≤67% in HAPC group.The plasma concentrations of CRP and SAA in HAPC patients were higher than those in health controls (P<0.05).CONCLUSION:Many kinds of differentially expressed proteins screened out are related to inflammation, which lays a foundation for further study of HAPC pathogenesis.%目的:应用串联质谱标签(TMT)结合质谱技术比较高原红细胞增多症(HAPC)患者和健康对照者血浆中的差异表达蛋白.方法:收集HAPC患者血浆4例,与之相匹配的健康对照组血浆5例,组内等量混合后利用多重免疫亲和层析柱去除14种高丰度蛋白,TMT试剂标记后样本进行液相色谱分离,采用HPLC-MS/MS质谱鉴定及相对定量;获取HAPC患者和健康对照血浆各20例,ELISA方法验证部分筛选的差异蛋白.结果:共鉴定和定量了1 094种蛋白,差异表达的蛋白有249种,其中HAPC组较健康对照组表达量≥1.5倍的上调蛋白质有162种,表达量≤67%的下调蛋白质有87种;C反应蛋白和血清淀粉样蛋白A在HAPC组中较健康对照组表达量均上调(P<0.05).结论:筛选出多种与炎症相关的差异表达蛋白,为深入研究HAPC发生机制奠定了良好基础.