title: A lipidomic screen of hyperglycemia-treated HRECs links 12/15-Lipoxygenase to microvascular dysfunction during diabetic retinopathy via NADPH oxidase.
abstract: Retinal hyperpermeability and subsequent macular edema is a cardinal feature of early diabetic retinopathy (DR). Here, we investigated the role of bioactive lipid metabolites, in particular 12/15-lipoxygenase (LOX)-derived metabolites, in this process. LC/MS lipidomic screen of human retinal endothelial cells (HRECs) demonstrated that 15-HETE was the only significantly increased metabolite (2.4 ± 0.4-fold, P = 0.0004) by high glucose (30 mM) treatment. In the presence of arachidonic acid, additional eicosanoids generated by 12/15-LOX, including 12- and 11-HETEs, were significantly increased. Fluorescein angiography and retinal albumin leakage showed a significant decrease in retinal hyperpermeability in streptozotocin-induced diabetic mice lacking 12/15-LOX compared with diabetic WT mice. Our previous studies demonstrated the potential role of NADPH oxidase in mediating the permeability effect of 12- and 15-HETEs, therefore we tested the impact of intraocular injection of 12-HETE in mice lacking the catalytic subunit of NADPH oxidase (NOX2). The permeability effect of 12-HETE was significantly reduced in NOX2(-/-) mice compared with the WT mice. In vitro experiments also showed that 15-HETE induced HREC migration and tube formation in a NOX-dependent manner. Taken together our data suggest that 12/15-LOX is implicated in DR via a NOX-dependent mechanism.
authors.affiliation: Oral Biology and Anatomy, College of Dental Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA; Ophthalmology and Culver Vision Discovery Institute, Medical College of Georgia, Georgia Regents University, Augusta, GA; Department of Clinical Biochemistry, Faculty of Pharmacy, Mansoura University, Mansoura, Egypt.
authors.forename: Ahmed S
authors.initials: AS
authors.lastname: Ibrahim
authors.affiliation: Oral Biology and Anatomy, College of Dental Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA.
authors.forename: Sally
authors.initials: S
authors.lastname: Elshafey
authors.affiliation: Department of Anesthesiology and Perioperative Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA.
authors.forename: Hassan
authors.initials: H
authors.lastname: Sellak
authors.affiliation: Oral Biology and Anatomy, College of Dental Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA; Ophthalmology and Culver Vision Discovery Institute, Medical College of Georgia, Georgia Regents University, Augusta, GA.
authors.forename: Khaled A
authors.initials: KA
authors.lastname: Hussein
authors.affiliation: Oral Biology and Anatomy, College of Dental Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA; Department of Anatomy, Faculty of Medicine,Mansoura University, Mansoura, Egypt. Electronic address: malshabrawey@gru.edu.
authors.forename: Mohamed
authors.initials: M
authors.lastname: El-Sherbiny
authors.affiliation: Department of Physiology, Medical College of Georgia, Georgia Regents University, Augusta, GA.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Abdelsaid
authors.affiliation: Biomedical Science Program, Faculty of Science, Qatar University, Doha, Qatar.
authors.forename: Nasser
authors.initials: N
authors.lastname: Rizk
authors.affiliation: Oral Biology and Anatomy, College of Dental Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA; Cellular Biology and Anatomy, Medical College of Georgia, Georgia Regents University, Augusta, GA.
authors.forename: Selina
authors.initials: S
authors.lastname: Beasley
authors.affiliation: Oral Biology and Anatomy, College of Dental Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA; Ophthalmology and Culver Vision Discovery Institute, Medical College of Georgia, Georgia Regents University, Augusta, GA; Cellular Biology and Anatomy, Medical College of Georgia, Georgia Regents University, Augusta, GA.
authors.forename: Amany M
authors.initials: AM
authors.lastname: Tawfik
authors.affiliation: Ophthalmology and Culver Vision Discovery Institute, Medical College of Georgia, Georgia Regents University, Augusta, GA; Cellular Biology and Anatomy, Medical College of Georgia, Georgia Regents University, Augusta, GA.
authors.forename: Sylvia B
authors.initials: SB
authors.lastname: Smith
authors.affiliation: Oral Biology and Anatomy, College of Dental Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA; Ophthalmology and Culver Vision Discovery Institute, Medical College of Georgia, Georgia Regents University, Augusta, GA; Department of Anatomy, Faculty of Medicine,Mansoura University, Mansoura, Egypt; Cellular Biology and Anatomy, Medical College of Georgia, Georgia Regents University, Augusta, GA.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Al-Shabrawey
mesh_terms: D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000235:genetics / Q000378:metabolism*; D001093:Arachidonate 15-Lipoxygenase / Q000235:genetics / Q000378:metabolism*; D003930:Diabetic Retinopathy / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000494:pharmacology*; D006943:Hyperglycemia / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism*
chemical_list: C475064:12-15-lipoxygenase; D006893:Hydroxyeicosatetraenoic Acids; D008562:Membrane Glycoproteins; D001092:Arachidonate 12-Lipoxygenase; D001093:Arachidonate 15-Lipoxygenase; C056558:CYBB protein, human; C488842:Cybb protein, mouse; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases
keywords: 12- and 15-HETEs; bioactive lipids; diabetic retinopathy; lipoxygenase; reduced nicotinamide adenine dinucleotide phosphate oxidase; retinal inflammation; retinal vascular leakage