title: UPLC-MS/MS-Based Profiling of Eicosanoids in RAW264.7 Cells Treated with Lipopolysaccharide.
abstract: While both the pro- and anti-inflammatory effects of several eicosanoids have been widely studied, the degree of inflammation in cells that results from various eicosanoids has yet to be comprehensively studied. The objective of this study was to assess the effect of lipopolysaccharide (LPS) treatment on eicosanoid content in RAW264.7 cells. An Ultra performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS)-based profiling method was used to analyze the eicosanoid contents of RAW264.7 cells treated with different LPS concentrations. The profiling data were subjected to statistical analyses, such as principal component analysis (PCA) and hierarchical clustering analysis. LPS treatment increased nitric oxide production and secretion of pro-inflammatory cytokines, such as tumor necrosis factor-α and interleukin-6, in a concentration-dependent manner. In total, 79 eicosanoids were identified in the cells. RAW264.7 cells treated with different LPS concentrations were well differentiated in the PCA score plot. A heatmap was used to identify the eicosanoids that were up- or down-regulated according to the degree of inflammation and LPS concentration. Thirty-nine eicosanoids were upregulated and seven were down-regulated by LPS treatment in a concentration-dependent manner. Our novel UPLC-MS/MS technique can profile eicosanoids, and can evaluate the correlations between inflammation and eicosanoid metabolism.
authors.affiliation: Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 369-873, Korea. jaewon3@gmail.com.
authors.forename: Jae Won
authors.initials: JW
authors.lastname: Lee
authors.affiliation: Department of Applied Chemistry, The Institute of Natural Science, College of Applied Science, Kyung Hee University, Yongin 446-701, Korea. hjmok@khu.ac.kr.
authors.forename: Hyuck Jun
authors.initials: HJ
authors.lastname: Mok
authors.affiliation: Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 369-873, Korea. dylee0809@korea.kr.
authors.forename: Dae-Young
authors.initials: DY
authors.lastname: Lee
authors.affiliation: Department of Applied Chemistry, The Institute of Natural Science, College of Applied Science, Kyung Hee University, Yongin 446-701, Korea. scpark1126@khu.ac.kr.
authors.forename: Seung Cheol
authors.initials: SC
authors.lastname: Park
authors.affiliation: Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 369-873, Korea. bms928@nate.com.
authors.forename: Myeong Soon
authors.initials: MS
authors.lastname: Ban
authors.affiliation: Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 369-873, Korea. jehun@korea.kr.
authors.forename: Jehun
authors.initials: J
authors.lastname: Choi
authors.affiliation: Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 369-873, Korea. pcg@korea.kr.
authors.forename: Chun Geon
authors.initials: CG
authors.lastname: Park
authors.affiliation: Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 369-873, Korea. ay21cay@korea.kr.
authors.forename: Young-Sup
authors.initials: YS
authors.lastname: Ahn
authors.affiliation: Department of Applied Chemistry, The Institute of Natural Science, College of Applied Science, Kyung Hee University, Yongin 446-701, Korea. kimkp@khu.ac.kr.
authors.forename: Kwang Pyo
authors.initials: KP
authors.lastname: Kim
authors.affiliation: Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 369-873, Korea. khd0303@rda.go.kr.|Department of Biochemistry, School of Life Sciences, Chungbuk National University, Cheongju 361-763, Korea. khd0303@rda.go.kr.
authors.forename: Hyung Don
authors.initials: HD
authors.lastname: Kim
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D015777:Eicosanoids / Q000032:analysis / Q000276:immunology*; D007249:Inflammation / Q000276:immunology; D015850:Interleukin-6 / Q000032:analysis / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology*; D008264:Macrophages / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D009569:Nitric Oxide / Q000032:analysis / Q000276:immunology; D000067996:RAW 264.7 Cells; D053719:Tandem Mass Spectrometry / Q000379:methods; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000276:immunology
chemical_list: D015777:Eicosanoids; D015850:Interleukin-6; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide
keywords: UPLC-MS/MS; eicosanoids; inflammation; lipopolysaccharide; macrophage