南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (12): 1409-1420.doi: 10.12122/j.issn.1673-4254.2019.12.04

• • 上一篇    下一篇

基于LC-MS筛选肺结核耐药患者的血清代谢标志物

林东子，王 威，邱 峰，李玉美，余晓琳，林炳耀，陈胤文，雷春燕，马 燕，曾今诚，周 杰   

  • 出版日期:2019-12-27 发布日期:2019-12-20
  • 基金资助:

Mass spectrometry-based identification of new serum biomarkers in patients with multidrug resistant pulmonary tuberculosis

  

  • Online:2019-12-27 Published:2019-12-20

HTML

PDF

1094

摘要： 目的 筛选潜在可用来指示不同耐药肺结核的新型血清代谢标志物，并分析其潜在机制与功能。方法 收集结核药敏（DS）、单耐药异烟肼（MR-INH）、单耐药利福平（MR-RFP）、耐多药（MDR）和多耐药（PR）患者血清样本。有机试剂振荡、去蛋白、提取全部血清样本中的代谢物质，LC-MS/MS 技术检测全部样本中的代谢物质。Pareto-scaling 方法进行归一化，应 用 MetaboAnalyst 4.0软件进行差异分析，组间T检验，P≤0.05视为有差异的代谢物。进一步通过代谢物功能富集及共表达分析，明确差异代谢物的功能并探讨其作为标志物的意义与机制。结果 基于LC-MS/MS技术分析发现相较于DS组，MR-INH组存在286个异常表达代谢物、MR-RFP组存在362个差异代谢物、MDR组存在277个差异代谢物、PR组存在1208个差异代谢物。其中Acetylagmatine（P<0.05）、Aminopentol（P<0.05）、Tetracosanyl oleate（P<0.05）在MR-INH组中表达具有显著差异；Ala His Pro Thr（P<0.001）、Glycinoprenol-9（P<0.05）在MR-RFP组中表达差异明显；Trimethylamine（P<0.05）、Penaresidin A（P< 0.05）、Verazine（P<0.05）在MDR组中表达具有显著差异；PIP（18∶1（11Z）/18∶3（6Z,9Z,12Z））（P<0.001）、Pro Arg Trp Tyr（P< 0.001）、N-Methyldioctylamine（P<0.001）、Phytolaccoside E（P<0.05）在PR组中表达具有显著差异。与DS组相比，Phthalic acid Mono-2-ethylhexyl Ester（P<0.05）和 Eicosanoyl-EA（P<0.05）在 MR-INH、MR- RFP、MDR、PR 组均存在显著差异。结论 Acetylagmatine、Aminopentol、Tetracosanyl oleate、Ala His Pro Thr、Glycinoprenol-9、Trimethylamine、Penaresidin A、Verazine、PIP（18∶1（11Z）/18∶3（6Z,9Z,12Z））、Pro Arg Trp Tyr、N-Methyldioctylamine、Phytolaccoside E、Phthalic acid Mono-2-ethylhexyl Ester、Eicosanoyl-EA共14种新型代谢物可能是指示结核单耐药、耐多药、多耐药的潜在标志物，联合分子标志物来评估结核耐药的发生发展，可以增强诊断的灵敏度和特异性，具有一定的临床应用前景。

Abstract: Objective To screen new serum metabolic biomarkers for different drug resistance profiles of pulmonary tuberculosis (TB) and explore their mechanisms and functions. Methods We collected serum samples from TB patients with drug sensitivity (DS), monoresistance to isoniazid (MR-INH), monoresistance to rifampin (MR-RFP), multidrug resistance (MDR), and polyresistance (PR). The metabolites in the serum samples were extracted by oscillatory and deproteinization for LC-MS/MS analysis, and the results were normalized by Pareto-scaling method and analyzed using Metaboanalyst 4.0 software to identify the differential metabolites. The differential metabolites were characterized by function enrichment and co-expression analysis to explore their function and possible pathological mechanisms. Results Compared with the DS group, 286 abnormally expressed metabolites were identified in MR-INH group, 362 in MR-RPF group, 277 in MDR group and 1208 in PR group by LC-MS/MS analysis. Acetylagmatine (P<0.05), aminopentol (P<0.05), and tetracosanyl oleate (P<0.05) in MR-INH group; Ala His Pro Thr (P<0.001) and glycinoprenol-9 (P<0.05) in MR-RFP group; trimethylamine (P<0.05), penaresidin A (P<0.05), and verazine (P<0.05) in MDR group; and PIP (18:1(11Z)/18:3(6Z,9Z,12Z)) (P<0.001), Pro Arg Trp Tyr (P<0.001), N-methyldioctylamine (P<0.001), and phytolaccoside E (P< 0.05) in PR group all showed significant differential expressions. Significant differential expressions of phthalic acid mono-2-ethylhexyl ester (P<0.05) and eicosanoyl-EA (P<0.05) were found in all the drug resistant groups as compared with DS group. Conclusion Acetylagmatine, aminopentol, tetracosanyl oleate, Ala His Pro Thr, glycinoprenol-9, trimethylamine, penaresidin A, verazine, PIP(18:1(11Z)/18:3(6Z,9Z,12Z)), Pro Arg Trp Tyr, N-methyldioctylamine, phytolaccoside E, phthalic acid mono-2-ethylhexyl ester, and eicosanoyl-EA are potentially new biomarkers that indicate monoresistance, multi-drug resistance and polyresistance of Mycobacterium tuberculosis. The combined use of these biomarkers potentially allows for assessment of drug resistance in TB and enhances the diagnostic sensitivity and specificity.