Search for Potential Biomarkers by UPLC/Q-TOF–MS Analysis of Dynamic Changes of Glycerophospholipid Constituents of RAW264.7 Cells Treated With NSAID
Search for Potential Biomarkers by UPLC/Q-TOF–MS Analysis of Dynamic Changes of Glycerophospholipid Constituents of RAW264.7 Cells Treated With NSAID
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DOI：
10.1007/s10337-014-2822-6
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发表时间：
2015-02
期刊：
影响因子：
1.7
通讯作者：
Xia Wu;Lilii Zhao;Haibo Peng;Yuqi She;Y. Feng
中科院分区：
文献类型：
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作者：
Xia Wu;Lilii Zhao;Haibo Peng;Yuqi She;Y. Feng
众所周知，组织中甘油磷脂（GPL）成分的改变可导致疾病。本文报道了利用现代分析技术和化学计量学方法揭示不同状态RAW264.7细胞GPL组分的动态变化，以寻找潜在的生物标志物。开发了Kdo 2-脂质A（KLA）刺激的RAW264.7细胞的炎症模型。通过使用UPLC-Q/TOF-MS分析来确定对用三种非甾体抗炎药（NSAID）（阿司匹林、吲哚美辛和布洛芬）治疗的模型的影响。获得了不同条件下代表GPL代谢的总离子电流曲线，并通过主成分分析（PCA）和偏最小二乘判别分析（PLSDA）处理数据。结果显示GPL代谢物的变化与NSAID的抗炎作用有关。通过使用ofttest、Shrinkt、主成分线性判别（PCLDA）、投影变量重要性（VIP）和PLSDA选择17个潜在的生物标志物。在这些生物标志物中，磷脂酰胆碱（PC，16：0/18：1）和磷脂酰乙醇胺（PE，18：0/18：1）的量在三个NSAID治疗组中显著变化，并且可能是炎症发生和消退的重要GPL生物标志物。基于UPLC/Q-TOF-MS的代谢组学为抗炎药物的作用机制提供了新的见解，与传统生物学研究不同。
It is well-known that alteration of the glycerophospholipid (GPL) constituents of tissues can lead to disease. We report dynamic changes of GPL constituents of RAW264.7 cells with different status, revealed by use of modern analytical techniques and chemometrics to find potential biomarkers. An inflammation model of Kdo2-lipid A (KLA)-stimulated RAW264.7 cells was developed. Effects on the model of treatment with three non-steroidal anti-inflammatory drugs (NSAID), aspirin, indomethacin, and brufen, were determined by use of UPLC-Q/TOF–MS analysis. Total-ion-current profiles representative of GPL metabolism under different conditions were acquired and the data were processed by principal-components analysis (PCA) and partial least-squares discriminant analysis (PLSDA). The results revealed changes of GPL metabolites related to the anti-inflammatory effects of the NSAID. Seventeen potential biomarkers were selected by use ofttests, Shrinkt, Principal Component Linear Discriminant (PCLDA), Variable importance in projection (VIP), and PLSDA. Among these biomarkers, amounts of phosphatidylcholine (PC, 16:0/18:1) and phosphatidylethanolamine (PE, 18:0/18:1) changed significantly in the three NSAID treatment groups and may be the important GPL biomarkers of the occurrence and resolution of inflammation. UPLC/Q-TOF–MS-based metabolomics provide novel insight into the mechanism of action of anti-inflammatory drugs distinct from that afforded by traditional biological investigations.