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Generation of Bioactive Oxylipins from Exogenously Added Arachidonic, Eicosapentaenoic and Docosahexaenoic Acid in Primary Human Brain Microvessel Endothelial Cells

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Lipids

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.

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Abbreviations

ARA:

Arachidonic acid (20:4n-6)

ALA:

α-Linolenic acid (18:3n-3)

BBB:

Blood–brain barrier

BSA:

Bovine serum albumin

COX:

Cyclooxygenase

CYP:

Cytochrome P450

DGLA:

Dihomo-γ-linolenic acid (20:3n-6)

DHA:

Docosahexaenoic acid (22:6n-3)

DiHDPE:

Dihydroxy-docosapentaenoic acid

DiHETE:

Dihydroxy-eicosatetraenoic acid

DiHETrE:

Dihydroxy-eicosatrienoic acid

DiHOME:

Dihydroxy-octadecenoic acid

EDA:

Eicosadienoic acid (20:2n-6)

EPA:

Eicosapentaenoic acid (20:5n-3)

EpDPE:

Epoxy-docosapentaenoic acid

EpODE:

Epoxy-octadecadienoic acid

EpOME:

Epoxy-octadecenoic acid

GLA:

γ-Linolenic acid (18:3n-6)

HBMEC:

Human brain microvessel endothelial cells

HDoHE:

Hydroxy-docosahexaenoic acid

HEPE:

Hydroxy-eicosapentaenoic acid

HETE:

Hydroxy-eicosatetraenoic acid

HETrE:

Hydroxy-eicosatrienoic acid

HHTrE:

Hydroxy-heptadecatrienoic acid

HODE:

Hydroxy-octadecadienoic acid

HOTrE:

Hydroxy-octadecatrienoic acid

HX:

Hepoxilin

LNA:

Linoleic acid (18:2n-6)

LOX:

Lipoxygenase

LT:

Leukotriene

LX:

Lipoxin

MEA:

Mead acid (20:3n-9)

MaR:

Maresin

NE:

Non enzymatic products

oxo-ETE:

Oxo-eicosatetraenoic acid

oxo-ODE:

Oxo-octadecadienoic acid

oxo-OTrE:

Oxo-octadecatrienoic acid

PD:

Protectin

PG:

Prostaglandin

PUFA:

Polyunsaturated fatty acid

Rv:

Resolvin

TriHOME:

Trihydroxy-octadecenoic acid

TX:

Thromboxane

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Acknowledgments

This work was supported by Natural Sciences and Engineering Research Council (NSERC) grants (to H.M.A, D.W.M, and G.M.H). G.M.H. is a Canada Research Chair in molecular cardiolipin metabolism.

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Authors and Affiliations

  1. Department of Human Nutritional Sciences, University of Manitoba, Winnipeg, Canada

    Harold M. Aukema & Tanja Winter

  2. Institute of Cardiovascular Sciences, St. Boniface Hospital Research Center, Winnipeg, Canada

    Amir Ravandi

  3. Departments of Pharmacology and Therapeutics, Faculty of Medicine, University of Manitoba, 753 McDermot Avenue, Winnipeg, MB, R3E 0W3, Canada

    Siddhartha Dalvi, Donald W. Miller & Grant M. Hatch

  4. Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, Canada

    Siddhartha Dalvi, Donald W. Miller & Grant M. Hatch

  5. Center for Research and Treatment of Atherosclerosis, University of Manitoba, Winnipeg, Canada

    Grant M. Hatch

  6. DREAM Children’s Hospital Research Institute of Manitoba, Winnipeg, MB, R3E 0T6, Canada

    Grant M. Hatch

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  1. Harold M. Aukema
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  2. Tanja Winter
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  4. Siddhartha Dalvi
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Correspondence to Grant M. Hatch.

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Aukema, H.M., Winter, T., Ravandi, A. et al. Generation of Bioactive Oxylipins from Exogenously Added Arachidonic, Eicosapentaenoic and Docosahexaenoic Acid in Primary Human Brain Microvessel Endothelial Cells. Lipids 51, 591–599 (2016). https://doi.org/10.1007/s11745-015-4074-0

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  • DOI: https://doi.org/10.1007/s11745-015-4074-0

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