基于非标记定量技术的继发性肺结核患者血浆蛋白质组学研究

Study on plasma proteomics of patients with secondary pulmonary tuberculosis based on label-free quantitative technology

目的研究继发性肺结核患者特异蛋白标志物的血浆蛋白质组学,寻找继发性肺结核特异性蛋白标志物。方法应用非标记(label-free)定量蛋白质组学技术检测30例继发性肺结核患者(结核组)和30名健康对照者(对照组)血浆蛋白质谱,结核组某蛋白丰度较对照组差异倍数>2(上调)或<0.5(下调)及两组某蛋白丰度经t检验P<0.05的蛋白被认为是差异蛋白。应用基因本体论(GO)、京都基因与基因组百科全书(KEGG)等分析软件对差异蛋白进行生物信息学分析。两组间蛋白平均丰度的差异分析采用t检验,以P<0.05为差异有统计学意义。结果结核组与对照组比较共发现518个差异表达蛋白,其中有256个差异蛋白表达上调,262个差异蛋白表达下调。生物信息学分析发现差异蛋白的分子功能主要集中在结合(29.79%,406/1363)和蛋白质结合(24.80%,338/1363);参与生物学过程主要为参与细胞过程(14.75%,467/3167)和代谢过程(11.11%,352/3167);差异蛋白的细胞定位分析发现,大部分蛋白定位在细胞内(22.07%,389/1763)及细胞(22.01%,388/1763)。KEGG分析发现背腹轴形成通路及黏着斑形成通路等17个上调通路与1个下调通路在结核组与对照组间具有差异性。结论结核组患者的血浆蛋白发生了明显的改变,这为探索继发性肺结核发生发展的机制提供了有价值的线索。

Objective To study the plasma proteomics of specific protein markers in patients with secondary pulmonary tuberculosis,to explore specific protein markers of secondary pulmonary tuberculosis.Methods Blood samples were obtained from 30 secondary pulmonary tuberculosis(tuberculosis group)and 30 healthy controls and a certain protein mass spectrometry was detected using label-free quantitative technology.If the protein abundance of the tuberculosis group more than 2(up-regulated)or<0.5(down-regulated)compared with that of the control group,and P<0.05 using t test,it was considered to be a differential protein bioinformatics analysis of differential proteins was performed using GO and KEGG.The difference in the average protein abundance between the two groups was analyzed by t test and P<0.05 was considered statistically significant.Results A total of 518 differential proteins were found in the two groups.Of them,256 proteins were up-regulated and 262 were downregulated.Bioinformatics analysis showed that the main functions of differential proteins were binding(29.79%,406/1363)and protein binding(24.80%,338/1363),and cell process(14.75%,467/3167)and metabolic process(11.11%,352/3167)were the main processes biological they involved.By cell location analysis,it was found that most of the proteins were located within the cell(22.07%,389/1763)and cell(22.01%,388/1763).KEGG analysis showed that 17 up-regulated pathways(such as dorsal ventral axis pathway and focal adhesion pathway)and 1 down regulated pathway were different between tuberculosis group and control group.Conclusion The plasma protein of patients with secondary pulmonary tuberculosis may have changed significantly,which provides valuable clues for exploring the mechanism of the secondary pulmonary tuberculosis.