LC-MS/MS法检测不明原因复发性流产患者蜕膜组织中多种氨基酸水平

Determination of multiple amino acids levels in decidua tissues of patients with unexplained recurrent spontaneous abortion by LC-MS/MS

目的分析不明原因复发性流产(URSA)患者与正常人工流产妇女蜕膜组织中7种氨基酸水平的差异,探讨URSA患者蜕膜组织中氨基酸的变化及临床意义。方法采用高效液相色谱串联质谱法(LC-MS/MS)检测15例URSA患者和20例人工流产妇女蜕膜组织中7种氨基酸水平的差异。组织样本充分研磨后加入含有同位素内标的甲醇溶液,添加比例为1∶9(M∶V),混匀并离心,提取100μL上清液进行检测。选用API3200型液相色谱串联质谱的多重反应监测(MRM)扫描方式。结果本方法简单、快速、高通量,且具有良好的重复性和稳定性。URSA患者蜕膜组织中鸟氨酸、甘氨酸水平均高于人工流产妇女(P<0.05);而两组间丙氨酸、缬氨酸、精氨酸、酪氨酸、瓜氨酸水平差异无统计学意义(P>0.05)。结论 URSA患者与人工流产妇女蜕膜组织中氨基酸水平存在差异。深入研究可能为不明原因复发流产的病因学提供基础。

Objective To analyze the difference of seven amino acids in decidua between unexplained recurrent spontaneous abortion(URSA)patients and normal induced abortion women,and to explore the changes and clinical significance of amino acids in decidua of URSA patients.Methods High performance liquid chromatography tandem mass spectrometry(LC-MS/MS)was used to detect the levels of seven amino acids in decidua of 15 URSA patients and 20 normal induced abortion women.The tissue samples were ground thoroughly and then added with methanol solution containing the internal standard of isotope.The ratio was 1∶9(M∶V),mixed and centrifuged.The supernatant of 100μL was extracted for detection.API3200 tandem mass spectrometry(MRM)scanning mode was used.Results This method was simple,rapid,high-throughput,and had good repeatability and stability.The levels of ornithine and glycine in decidua of URSA patients were higher than those of normal induced abortion women(P<0.05),but the levels of other amino acids(Ala,Val,Arg,Tyr,Cit)were not significantly different(P>0.05).Conclusion There were differences of amino acid levels in decidua tissues between normal induced abortion women and URSA patients.Further studies may provide the basis for the etiology of recurrent spontaneous abortion.