Araujo P., Lucena E., Yang Y., Ceemala B., Mengesha Z., Holen E.
National Institute of Nutrition and Seafood Research (NIFES), PO Box 2029 Nordnes, N-5817 Bergen, Norway; Centro de Biofísica y Bioquímica, Laboratorio de Fisiología Celular, Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela; Department of Chemistry, University of Bergen, PO Box 7803, N-5020 Bergen, Norway; Department of Industrial Chemistry, Bahir Dar University, PO Box 79, Bahir Dar, Ethiopia
Araujo, P., National Institute of Nutrition and Seafood Research (NIFES), PO Box 2029 Nordnes, N-5817 Bergen, Norway; Lucena, E., National Institute of Nutrition and Seafood Research (NIFES), PO Box 2029 Nordnes, N-5817 Bergen, Norway, Centro de Biofísica y Bioquímica, Laboratorio de Fisiología Celular, Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela; Yang, Y., National Institute of Nutrition and Seafood Research (NIFES), PO Box 2029 Nordnes, N-5817 Bergen, Norway, Department of Chemistry, University of Bergen, PO Box 7803, N-5020 Bergen, Norway; Ceemala, B., National Institute of Nutrition and Seafood Research (NIFES), PO Box 2029 Nordnes, N-5817 Bergen, Norway; Mengesha, Z., National Institute of Nutrition and Seafood Research (NIFES), PO Box 2029 Nordnes, N-5817 Bergen, Norway, Department of Chemistry, University of Bergen, PO Box 7803, N-5020 Bergen, Norway, Department of Industrial Chemistry, Bahir Dar University, PO Box 79, Bahir Dar, Ethiopia; Holen, E., National Institute of Nutrition and Seafood Research (NIFES), PO Box 2029 Nordnes, N-5817 Bergen, Norway
The production of prostaglandins (PGE2, PGE3) and leukotrienes (LTB4, LTB5) in salmon head kidney cell cultures, exposed to different combinations of 20:4ω-6, 20:5ω-3 and 22:6ω-3 polyunsaturated fatty acids (PUFAs), was evaluated by means of a two level factorial design and LC-MS/MS. The method was selective for the pro- and anti-inflammatory analytes and their corresponding stable-isotope labelled internal standards. The regression models were linear over the concentration range 0.5-150ng/ml with limits of detection of 0.25ng/ml and quantification of 0.40ng/ml for the analysed metabolites. The recovery ranged from 78 to 107% for prostaglandins and 73 to 115% for leukotrienes. The analysis of the samples exposed to different combinations of PUFAs revealed that the presence of single ω-3 PUFAs brought an enhancement of the metabolites from the lipooxygenase pathway, specially LTB4, and a reduction of the metabolites from the cyclooxygenase pathway (PGE2 and PGE3), while the two-term interactions generated the opposite effect (high concentration of prostaglandins and low concentrations of leukotrienes). To our knowledge, this is the first implementation of a fully crossed design for investigating the impact of ω-6 and ω-3 PUFAs on the production of eicosanoids not only through their individual but also through their combined effects on Atlantic salmon head kidney cells. © 2014 Elsevier B.V.
Biomolecules; Design of experiments; Isotopes; Liquid chromatography; Mass spectrometry; Metabolites; Polyunsaturated fatty acids; Regression analysis; Eicosanoids; Leukotrienes; Liquid chromatography-tandem mass spectrometry; Prostaglandins; Salmon heads; Cell culture; icosanoid; leukotriene B4; leukotriene B5; lipoxygenase; omega 3 fatty acid; omega 6 fatty acid; prostaglandin E2; prostaglandin E3; prostaglandin synthase; leukotriene; omega 3 fatty acid; omega 6 fatty acid; prostaglandin; animal cell; article; Atlantic salmon; cell culture; controlled study; factorial design; head kidney; incubation time; intervention study; isotope labeling; kidney cell; limit of detection; liquid chromatography; metabolite; nonhuman; priority journal; tandem mass spectrometry; animal; Atlantic salmon; chemistry; evaluation study; head kidney; liquid chromatography; metabolism; procedures; tandem mass spectrometry; Salmo salar; Animals; Chromatography, Liquid; Fatty Acids, Omega-3; Fatty Acids, Omega-6; Head Kidney; Leukotrienes; Limit of Detection; Prostaglandins; Salmo salar; Tandem Mass Spectrometry