High-throughput lipidomic analysis of fatty acid derived eicosanoids and N-acylethanolamines

Dumlao, Darren S.; Buczynski, Matthew W.; Norris, Paul C.; Harkewicz, Richard; Dennis, Edward A.

doi:10.1016/j.bbalip.2011.06.005

Cited by 130 publications

(103 citation statements)

References 35 publications

Supporting

Mentioning

100

Contrasting

Unclassified

Order By: Relevance

“…Eicosanoids were analyzed by a stable isotope dilution LC/MS method utilizing 26 deuterated internal standards ( 28,29 ). The metabolites were quantifi ed after separation by reverse phase chromatography on a 2.1 × 100 mm BEH Shield column, 1.7 M (Waters, Milford, MA) employing an Acquity UPLC system (Waters).…”

Section: Rna-seq and Snp Arraysmentioning

confidence: 99%

Biomarkers of NAFLD progression: a lipidomics approach to an epidemic

Gorden

Myers

Ivanova

et al. 2015

Journal of Lipid Research

Self Cite

288

266

View full text Add to dashboard Cite

Nonalcoholic fatty liver disease (NAFLD) is rapidly becoming one of the most common forms of liver disease in Abstract The spectrum of nonalcoholic fatty liver disease (NAFLD) includes steatosis, nonalcoholic steatohepatitis (NASH), and cirrhosis. Recognition and timely diagnosis of these different stages, particularly NASH, is important for both potential reversibility and limitation of complications. Liver biopsy remains the clinical standard for defi nitive diagnosis. Diagnostic tools minimizing the need for invasive procedures or that add information to histologic data are important in novel management strategies for the growing epidemic of NAFLD. We describe an "omics" approach to detecting a reproducible signature of lipid metabolites, aqueous intracellular metabolites, SNPs, and mRNA transcripts in a double-blinded study of patients with different stages of NAFLD that involves profi ling liver biopsies, plasma, and urine samples. Using linear discriminant analysis, a panel of 20 plasma metabolites that includes glycerophospholipids, sphingolipids, sterols, and various aqueous small molecular weight components involved in cellular metabolic pathways, can be used to differentiate between NASH and steatosis. This identifi cation of differential biomolecular signatures has the potential to improve clinical diagnosis and facilitate therapeutic intervention of

show abstract

Section: Rna-seq and Snp Arraysmentioning

confidence: 99%

Biomarkers of NAFLD progression: a lipidomics approach to an epidemic

Gorden

Myers

Ivanova

et al. 2015

Journal of Lipid Research

Self Cite

288

266

View full text Add to dashboard Cite

show abstract

“…PE and PI molecular species were identified by multiple reaction monitoring experiments on chromatographic effluent by comparison with previously published data (24,(26)(27)(28)(29)(30)(52)(53)(54). Cutoff parameter was set at m/z 150, and fragmentation amplitude was set at one arbitrary unit.…”

Section: Immunoblotmentioning

confidence: 99%

Group V Secreted Phospholipase A2 Is Upregulated by IL-4 in Human Macrophages and Mediates Phagocytosis via Hydrolysis of Ethanolamine Phospholipids

Rubio

Rodríguez

Gil-de-Gómez

et al. 2015

The Journal of Immunology

View full text Add to dashboard Cite

Studies on the heterogeneity and plasticity of macrophage populations led to the identification of two major polarization states: classically activated macrophages or M1, induced by IFN-γ plus LPS, and alternatively activated macrophages, induced by IL-4. We studied the expression of multiple phospholipase A2 enzymes in human macrophages and the effect that polarization of the cells has on their levels. At least 11 phospholipase A2 genes were found at significant levels in human macrophages, as detected by quantitative PCR. None of these exhibited marked changes after treating the cells with IFN-γ plus LPS. However, macrophage treatment with IL-4 led to strong upregulation of the secreted group V phospholipase A2 (sPLA2-V), both at the mRNA and protein levels. In parallel with increasing sPLA2-V expression levels, IL-4–treated macrophages exhibited increased phagocytosis of yeast-derived zymosan and bacteria, and we show that both events are causally related, because cells deficient in sPLA2-V exhibited decreased phagocytosis, and cells overexpressing the enzyme manifested higher rates of phagocytosis. Mass spectrometry analyses of lipid changes in the IL-4–treated macrophages suggest that ethanolamine lysophospholipid (LPE) is an sPLA2-V–derived product that may be involved in regulating phagocytosis. Cellular levels of LPE are selectively maintained by sPLA2-V. By supplementing sPLA2-V–deficient cells with LPE, phagocytosis of zymosan or bacteria was fully restored in IL-4–treated cells. Collectively, our results show that sPLA2-V is required for efficient phagocytosis by IL-4–treated human macrophages and provide evidence that sPLA2-V–derived LPE is involved in the process.

show abstract

“…All plasma samples were stored at Ϫ 80°C, thawed once, and immediately used for free fatty acid and eicosanoid isolation as described previously ( 15,17 ). Briefl y, 50 µl plasma was spiked with a cocktail of 26 deuterated internal standards (individually purchased from Cayman Chemicals, Ann Arbor, MI) and brought to a volume of 1 ml with 10% methanol.…”

Section: Lipid Extractionmentioning

confidence: 99%

“…Briefl y, 50 µl plasma was spiked with a cocktail of 26 deuterated internal standards (individually purchased from Cayman Chemicals, Ann Arbor, MI) and brought to a volume of 1 ml with 10% methanol. The samples were then purifi ed by solid phase extraction on Strata-X columns (Phenomenex, Torrance, CA), using an activation procedure consisting of consecutive washes lipidomics techniques, our laboratory has developed a robust and comprehensive approach to the lipidomics analysis of hundreds of fatty acids, acylethanolamines, and infl ammatory eicosanoids, including their numerous metabolites arising from an array of cyclooxygenases, lipoxygenases, cytochrome P450s, and nonenzymatic oxidation-producing isoprostanes, as well as combinations thereof ( 15 ). Particular attention has been focused on the eicosanoids derived from arachidonic acid (AA), and we can now routinely quantify >150 such metabolites and have used this approach to profi le AA and other PUFAs as well as their metabolites in human plasma (15)(16)(17).…”

Section: Lipid Extractionmentioning

confidence: 99%