The impact of exogenous ω-6 and ω-3 polyunsaturated fatty acids on the induced production of pro- and anti-inflammatory prostaglandins and leukotrienes in Atlantic salmon head kidney cells using a full factorial design and LC–MS/MS

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2014-08, Vol.964, p.164-171
• Main Authors: Araujo, Pedro, Lucena, Eva, Yang, Yang, Ceemala, Bupesh, Mengesha, Zebasil, Holen, Elisabeth
• Format: Article
• Language: English
• Subjects: Animals; Chromatography, Liquid - methods; Eicosanoids; Experimental design; Factorial design; Fatty acids; Fatty Acids, Omega-3 - administration & dosage; Fatty Acids, Omega-3 - metabolism; Fatty Acids, Omega-6 - administration & dosage; Fatty Acids, Omega-6 - metabolism; Head Kidney - chemistry; Head Kidney - metabolism; Kidneys; Leukotrienes; Leukotrienes - analysis; Leukotrienes - metabolism; Limit of Detection; Liquid Chromatography Tandem Mass Spectrometry; Metabolites; Pathways; Polyunsaturated fatty acids; Prostaglandins; Prostaglandins - analysis; Prostaglandins - metabolism; Salmo salar - metabolism; Salmon; Salmon head kidney cell cultures; Tandem Mass Spectrometry - methods
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2014.01.018

•Fully crossed design for investigating the impact of ω-6 and ω-3 polyunsaturated fatty acids (PUFAs) on production of eicosanoids.•The individual PUFAs and their interactions are considered.•Probable eicosanoids’ signalling mechanism is suggested based on their observed behaviour. 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.