基于同位素标记的相对和绝对定量的糖尿病并发肺结核患者血浆蛋白质组学研究

目的: 比较糖尿病(diabetes mellitus,DM)患者与糖尿病并发肺结核(diabetes mellitus complicated with pulmonary tuberculosis,DM-PTB)患者的血浆蛋白质谱差异,筛选DM-PTB潜在蛋白质生物标志物,为进一步探索DM-PTB的发病机制提供思路。方法: 采用回顾性研究的方法,选取于2017年1月至2018年1月在黑龙江省传染病防治院确诊的DM患者和DM-PTB患者各16例,采用基于同位素标记的相对和绝对定量(isobaric tags for relative and absolute quantitation,iTRAQ)蛋白质组学技术分析两组间血浆蛋白质谱,以DM-PTB组对DM组蛋白表达量变化倍数>1.2或<0.83且P<0.05为标准筛选差异蛋白,并对差异蛋白进行基因本体(Gene Ontology,GO)功能注释和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路分析等生物信息学分析。结果: 共筛选出275个差异表达蛋白,其中有111个在DM-PTB组上调,有164个在DM-PTB组下调。GO分析显示,差异表达蛋白富集数目最多的生物学过程为细胞过程(92.00%,253/275),定位于细胞器(87.27%,240/275),分子功能为结合(90.55%,249/275)。KEGG代谢通路分析显示,差异表达蛋白共富集到26条代谢通路(错误发现率<0.05),富集数目最多的通路为黏着斑通路(22个蛋白),此外PI3K-Akt信号通路、胆固醇代谢通路、血小板激活通路、吞噬体通路和补体及凝血通路等与糖脂代谢和免疫相关的通路也发生了变化。结论: 本研究通过iTRAQ蛋白质组学方法筛选出了DM-PTB患者的差异表达蛋白,发现这些差异表达蛋白参与了与细胞活动、糖脂代谢和免疫调节相关的信号通路,提示上述通路的变化可能与DM-PTB的发病密切相关。

Plasma proteomics analysis of diabetic patients complicated with pulmonary tuberculosis by iTRAQ technology

Objective: To analyze the difference in plasma protein mass spectrum between diabetic (DM) patients and diabetic patients complicated with pulmonary tuberculosis (DM-PTB), screen potential protein biomarkers of DM-PTB, and to provide basis for further pathogenesis exploration. Methods: A retrospective study was conducted in 16 DM and 16 DM-PTB patients from Infectious Disease Hospital of Heilongjiang Province between January 1, 2017and January 1, 2018. The plasma protein mass spectra between the two groups were analyzed by isobaric tags for relative and absolute quantitation (iTRAQ) proteomics technology. The differential proteins were screened according to the following criteria: fold of protein expression change in DM-PTB compared to DM>1.2 or <0.83, and P<0.05. The differential proteins were analyzed by bioinformatics, such as Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and so on. Results: A total of 275 differentially expressed proteins were selected out, among which 111 were up-regulated and 164 were down-regulated in DM-PTB. GO analysis showed that the biological process with the largest number of differentially expressed proteins was cellular process (92.00%, 253/275), located in organelle (87.27%, 240/275) and the molecular function was binding (90.55%, 249/275). KEGG pathway analysis showed that the differentially expressed proteins were enriched into 26 pathways (false discovery rate <0.05) and focal adhesion was the pathway with the largest number of enrichment (22 proteins). Moreover, changes were also found in PI3K-Akt signaling pathway, cholesterol metabolism, platelet activation, phagosome, complement and coagulation pathways, et al., which related to glucose and lipid metabolism and immunity. Conclusion: In this study, differentially expressed proteins of DM-PTB patients were screened by iTRAQ quantitative proteomics, and they were involved in signaling pathways related to cellular activity, glucose and lipid metabolism, and immune regulation, suggesting that the changes of pathways mentioned above may be closely related to the pathogenesis of DM-PTB.