基于UPLC-QTOF/MS的尘肺潜伏性结核患者血清非靶向代谢组学分析

Serum metabolomics in latent pneumoconiosis tuberculosis patients based on ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry

目的:对尘肺潜伏性结核患者血清进行非靶向代谢组学分析,探索尘肺潜伏性结核感染的生物标志物。方法:于2018年12月,以北京某医院住院煤工尘肺患者39人为研究对象,采用结核分歧杆菌γ-干扰素体外释放试验(IGRAs)进行潜伏性结核筛查。根据筛查结果选择IGRAs阳性患者21人为尘肺潜伏性结核组,阴性患者18人为尘肺组,采用超高效液相色谱-四极杆飞行时间串联质谱(UPLC-QTOF/MS)技术对极性小分子代谢产物进行检测分析,数据采用Progenesis QI软件处理后进行多元统计分析,差异性代谢物结构鉴定采用精确质量数匹配和二级谱图匹配的方式,检索人类代谢组数据库(HMDB),将差异性代谢物导入MetaboAnalyst 4.0网站进行代谢通路分析。结果:共筛选出42种差异代谢物;排除外源性代谢物,共鉴定出14种内源性差异代谢产物。与尘肺组患者相比,尘肺潜伏性结核组患者体内PC[18∶4(6Z,9Z,12Z,15Z)/P-18∶1(11Z)]、3-Oxododecanoyl-CoA等6种代谢物上调;Stearoyl-CoA、(2S)-Pristanoyl-CoA等8种代谢物下调,可能与磷脂代谢、脂肪酸代谢、花生四烯酸代谢等通路相关。结论:尘肺潜伏性结核患者与普通尘肺患者的血清代谢组学特征存在差异,可为尘肺潜伏性结核感染诊断生物标志物研究提供参考。

Objective To explore the non-target metabonomics of serum in worker's pneumoconiosis(CWP)patients with latent tuberculosis and the biomarkers of latent tuberculosis infection of pneumoconiosis.Methods In December 2018,39 CWP inpatients from a hospital in Beijing were taken as subjects.The subjects were screened for latent tuberculosis using the in vitro release test of mycobacterium tuberculosis-interferon(IGRAs)test.According to the screening results,21 positive patients with latent tuberculosis infection were selected as the latent tuberculosis group of pneumoconiosis.While 18 negative patients with CWP alone were selected as the pneumoconiosis group.Polarity components of metabolites were analyzed by UPLC-QTOF/MS.The data was processed with Progenesis QI software for multidimensional statistical analysis.Identification of structure of differential metabolites were matched through accurate mass and secondary mass spectrum.Searching the Human Metabolome Database(HMDB),differential metabolites were imported into MetaboAnalyst 4.0 to analyze the metabolic pathways.Results All 42 differential metabolites were screened out.Excepted for exogenous metabolites,14 endogenous differential metabolites were identified.Compared with the pneumoconiosis group,6 metabolites including PC[18∶4(6Z,9Z,12Z,15Z)/P-18∶1(11Z)],3-Oxododecanoyl-CoA in the latent tuberculosis group were up-regulated,while 8 metabolites including the Stearoyl-CoA,(2S)-Pristanoyl-CoA were down-regulated.These results might be related to lipid,fatty acid and arachidonic acid metabolism pathways.Conclusion There are significant differences in serum metabonomics between the patients with latent tuberculosis of pneumoconiosis and the patients with ordinary pneumoconiosis,which provide a reference for the study of biomarkers for the diagnosis of latent tuberculosis infection of pneumoconiosis.