title: Generation of Bioactive Oxylipins from Exogenously Added Arachidonic, Eicosapentaenoic and Docosahexaenoic Acid in Primary Human Brain Microvessel Endothelial Cells.
abstract: The human blood-brain barrier (BBB) is the restrictive barrier between the brain parenchyma and the circulating blood and is formed in part by microvessel endothelial cells. The brain contains significant amounts of arachidonic acid (ARA), and docosahexaenoic acid (DHA), which potentially give rise to the generation of bioactive oxylipins. Oxylipins are oxygenated fatty acid metabolites that are involved in an assortment of biological functions regulating neurological health and disease. Since it is not known which oxylipins are generated by human brain microvessel endothelial cells (HBMECs), they were incubated for up to 30 min in the absence or presence of 0.1-mM ARA, eicosapentaenoic acid (EPA) or DHA bound to albumin (1:1 molar ratio), and the oxylipins generated were examined using high performance liquid chromatography-tandem mass spectrometry (HPLC/MS/MS). Of 135 oxylipins screened in the media, 63 were present at >0.1 ng/mL at baseline, and 95 were present after incubation with fatty acid. Oxylipins were rapidly generated and reached maximum levels by 2-5 min. While ARA, EPA and DHA each stimulated the production of oxylipins derived from these fatty acids themselves, ARA also stimulated the production of oxylipins from endogenous 18- and 20-carbon fatty acids, including α-linolenic acid. Oxylipins generated by the lipoxygenase pathway predominated both in resting and stimulated states. Oxylipins formed via the cytochrome P450 pathway were formed primarily from DHA and EPA, but not ARA. These data indicate that HBMECs are capable of generating a plethora of bioactive lipids that have the potential to modulate BBB endothelial cell function.
authors.affiliation: Department of Human Nutritional Sciences, University of Manitoba, Winnipeg, Canada.
authors.forename: Harold M
authors.initials: HM
authors.lastname: Aukema
authors.affiliation: Department of Human Nutritional Sciences, University of Manitoba, Winnipeg, Canada.
authors.forename: Tanja
authors.initials: T
authors.lastname: Winter
authors.affiliation: Institute of Cardiovascular Sciences, St. Boniface Hospital Research Center, Winnipeg, Canada.
authors.forename: Amir
authors.initials: A
authors.lastname: Ravandi
authors.affiliation: Departments of Pharmacology and Therapeutics, Faculty of Medicine, University of Manitoba, 753 McDermot Avenue, Winnipeg, MB, R3E 0W3, Canada.|Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, Canada.
authors.forename: Siddhartha
authors.initials: S
authors.lastname: Dalvi
authors.affiliation: Departments of Pharmacology and Therapeutics, Faculty of Medicine, University of Manitoba, 753 McDermot Avenue, Winnipeg, MB, R3E 0W3, Canada.|Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, Canada.
authors.forename: Donald W
authors.initials: DW
authors.lastname: Miller
authors.affiliation: Departments of Pharmacology and Therapeutics, Faculty of Medicine, University of Manitoba, 753 McDermot Avenue, Winnipeg, MB, R3E 0W3, Canada. GHatch@chrim.ca.|Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, Canada. GHatch@chrim.ca.|Center for Research and Treatment of Atherosclerosis, University of Manitoba, Winnipeg, Canada. GHatch@chrim.ca.|DREAM Children's Hospital Research Institute of Manitoba, Winnipeg, MB, R3E 0T6, Canada. GHatch@chrim.ca.
authors.forename: Grant M
authors.initials: GM
authors.lastname: Hatch
mesh_terms: D016718:Arachidonic Acid / Q000378:metabolism*; D001921:Brain / Q000098:blood supply* / Q000378:metabolism; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004281:Docosahexaenoic Acids / Q000378:metabolism*; D015118:Eicosapentaenoic Acid / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D055806:Microvessels / Q000166:cytology* / Q000378:metabolism; D054883:Oxylipins / Q000032:analysis / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D054883:Oxylipins; D004281:Docosahexaenoic Acids; D016718:Arachidonic Acid; D015118:Eicosapentaenoic Acid
keywords: Analytical techniques; Fatty atacids; General area; Lipid analysis; Lipid biochemistry; Mass spectrometry (MS); Metabolism; Oxidized lipids; Specific lipids; Synthesis