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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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Abstract

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 of t tests, 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.

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Acknowledgments

This work was supported by the Natural Science Foundation of China (nos. 21275036 and 81202429).

Conflict of interest

We declare that all authors have read the manuscript conscientiously, and all agree with its publication. The authors have no conflict of interest to report.

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Authors and Affiliations

  1. Center Laboratory, Guangdong Pharmaceutical University, Guangzhou, 510006, People’s Republic of China

    Xia Wu, Lili Zhao, Haibo Peng, Yuqi She & Yifan Feng

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  1. Xia Wu
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  2. Lili Zhao
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  3. Haibo Peng
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  4. Yuqi She
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  5. Yifan Feng
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Correspondence to Yifan Feng.

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Wu, X., Zhao, L., Peng, H. et al. Search for Potential Biomarkers by UPLC/Q-TOF–MS Analysis of Dynamic Changes of Glycerophospholipid Constituents of RAW264.7 Cells Treated With NSAID. Chromatographia 78, 211–220 (2015). https://doi.org/10.1007/s10337-014-2822-6

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  • DOI: https://doi.org/10.1007/s10337-014-2822-6

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