稳定同位素iTRAQ标记联合液相色谱-串联质谱法定量分析药物性肝损伤患者血清氨基酸的变化

Quantitative analysis of serum amino acids changes in patients with drug-induced liver injury by stable isotope iTRAQ labeling combined with liquid chromatography tandem mass spectrometry

目的探索药物性肝损伤(DILI)患者治疗的不同阶段血清氨基酸水平的变化差异,探讨其临床意义及可用于DILI早期诊断及疗效评价的氨基酸类生物标志物。方法收集21例健康志愿者(健康组)及24例DILI患者不同阶段(初诊、治疗、转归)的血清标本,采用稳定同位素iTRAQ标记方法联合液相色谱串联质谱技术检测血清氨基酸进行定量并分析其差异。结果在血清中共定量氨基酸28种,与健康组比较,DILI患者共有14种氨基酸浓度在诊疗不同阶段发生显著性改变;其中异亮氨酸、亮氨酸、苯丙氨酸及丝氨酸的浓度在初诊时显著升高,随治疗进程降低,逐渐接近于正常水平。结论 4种氨基酸中,苯丙氨酸与DILI的相关性更高,可能为DILI早期诊断及治疗效果评价的潜在生物标志物。

Objective To explore the changes of serum amino acid levels in the patients with drug-induced liver injury（DILI）at different stages,and then to investigate their clinical significance and the biomarkers available for early diagnosis and curative evaluation of DILI.Methods Serum samples from 21 healthy volunteers and 24 patients with different stages（first visiting,treatment,outcome）of DILI were collected.The stable isotope iTRAQ labeling combined with liquid chromatography tandem mass spectrometry was adopted to quantitatively detect the serum amino acids and analyzes their differences.Results There were quantified 28 amino acids in serum.Compared with the control group,14 kinds of amino acids had significant changes in concentrations during the different diagnosis and treatment periods;among them,the concentrations of isoleucine,leucine,phenylalanine and serine were significantly increased at first visiting,decreased with the treatment process and gradually close to the normal level.Conclusion Among 4kinds of amino acids,The correlation between phenylalanine and DILI is higher,which might be a potential biomarker for early diagnosis and therapeutic effect evaluation of DILI.