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高效液相色谱-质谱法对冠心病患者血清的脂质组学研究及其临床意义
          
HPLC-MS Study on Lipidomics for Coronary Artery Disease Patients' Serum and Its Significance in Clinical Medicine

摘    要
采用高效液相色谱-质谱法(HPLC-MS)对冠状动脉慢性完全堵塞(CTO)患者在不同患病阶段的血清试样进行了脂质组学研究。试验采集了CTO疑似人群、CTO患者及CTO患者经皮冠状动脉介入治疗(PCI)手术后24 h和72 h的血清,分别采用氯仿和体积比为25:75的乙酸乙酯-异辛烷混合溶液提取脂质,离心分离后,上清液用于Shotgun法(MS)和多反应监测(MRM)法(HPLC-MS)分析。样本直接进样电喷雾(ESI)质谱分析,用于Shotgun法进行广谱的脂质组学分析鉴定。通过Analyst 1.6软件提取质谱数据,并通过生物信息学工具“Lipid MS Predict”确认脂质分子,总共得到1 504种脂质分子,并利用MRM法确认其中有临床意义的631个脂质分子。通过生物信息学分析,得到对于CTO具有诊断价值的包括胆固醇酯CE 16:1和溶血磷脂酰乙醇胺LPE 18:1在内的9个标志性脂质分子。结合临床回访结果和脂质组学研究数据证实,CTO患者手术治疗后的预后情况和磷脂酸、磷脂酰胆碱、胆固醇酯等3类脂质的含量变化有关。确定的脂质标志物应用于临床诊断早期冠心病和评估手术治疗后的风险,具有显著而明确的临床医学价值。
标    签 高效液相色谱-质谱法   脂质组学   冠状动脉慢性完全堵塞   临床诊断   HPLC-MS   lipidomics   chronic complete coronary occlusion   clinical diagnosis  
 
Abstract
HPLC-MS was established to study the lipidomics of serum samples for the patients with chronic complete coronary occlusion (CTO) at the different stages of disease. Patients' serum of suspected CTO, CTO, and CTO after 24 h and 72 h of percutaneous coronary intervention (PCI) surgery were collected respectively in the test. The lipid in samples was extracted with chloroform and ethyl acetate-isooctane with volume ratio of 25:75, respectively. After centrifugation, the supernatant was taken for Shotgun method (MS) and multiple reaction monitoring (MRM) method (HPLC-MS). The supernatant sample was injected directly to electrospray (ESI) mass spectrometry for a broad-lipidomics analysis and identification by Shotgun method. The MS data was indexed and analyzed by software Analyst 1.6, and the lipid molecules were confirmed by the bioinformatics tool “Lipid MS Predict”. A total of 1 504 lipid molecules were identified, and among them, 631 lipid molecules with clinical significance were confirmed by using the MRM method. Through bioinformatics analysis, 9 lipid molecules with significance in CTO diagnosis at early stage were ascertained as the biomarker, including cholesterol ester (CE) 16:1 and lysophosphatidylethanolamine (LPE) 18:1. Feedback from the clinical return visit as well as research data confirmed that the postoperative risk prognosis of CTO patients was correlated positively with the increase of content of 3 lipid molecules including phosphatidic acid, phosphatidylcholine, and cholesterol ester. The lipid biomarkers found in the study can be applied to the clinical diagnosis of coronary heart disease at early stage and assessment of the risk after surgery, with definite significance and clear clinical medical value.

中图分类号 O657.63   DOI 10.11973/lhjy-hx202101006

 
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基金项目 复旦大学附属中山医院青年科学基金(2020ZSQN30)

收稿日期 2020/11/11

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备注王真昕,博士,主要从事临床检验工作,wang.zhenx-in@zs-hospital.sh.cn

引用该论文: WANG Zhenxin,SHEN Huali,YANG Pengyuan. HPLC-MS Study on Lipidomics for Coronary Artery Disease Patients' Serum and Its Significance in Clinical Medicine[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2021, 57(1): 32~35
王真昕,申华莉,杨芃原. 高效液相色谱-质谱法对冠心病患者血清的脂质组学研究及其临床意义[J]. 理化检验-化学分册, 2021, 57(1): 32~35


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