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

发布时间：2018-03-23 00:08

本文选题：高原红细胞增多症　切入点：串联质谱标签　出处：《中国病理生理杂志》2017年05期 　论文类型：期刊论文

【摘要】：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(P0.05)。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。
[Abstract]:Objective: to compare the differentially expressed proteins in plasma of patients with high altitude polycythemia (HAP) and healthy controls by tandem mass spectrometry (tandem mass spectrometry) and mass spectrometry. Methods: plasma samples from 4 patients with HAPC and 5 healthy controls were collected. The samples labeled with 14 kinds of high abundance protein TMT reagent were separated by HPLC-MS/MS mass spectrometry and relative quantitative analysis after the same amount of mixture was used to remove 14 kinds of high abundance protein TMT reagent by multiple immunoaffinity chromatography column. A total of 1 094 proteins were identified and quantified, and 249 differentially expressed proteins were identified and quantified by Elisa in 20 patients with HAPC and 20 healthy control plasma samples. Among them, there were 162 kinds of up-regulated proteins in HAPC group, which were more than 1.5 times higher than those in healthy control group. 87 C-reactive proteins and serum amyloid A were up-regulated in HAPC group compared with healthy control group. Conclusion: a variety of differentially expressed proteins related to inflammation were screened. It lays a good foundation for further study on the mechanism of HAPC.
【作者单位】：青海大学医学院;青海省心脑血管病专科医院内科;青海大学医学院高原医学研究中心;青海省高原医学应用基础重点实验室;青海-犹他高原医学联合重点实验室;
【基金】：青海省科技厅科技支撑计划(No.2015-SF-124);青海省科技厅应用基础基金项目(No.2016-ZJ-706)
【分类号】：R339.5+4;;R599.9

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