A lipidomic screen of hyperglycemia-treated hrecs links 12/15-lipoxygenase to microvascular dysfunction during diabetic retinopathy via NADPH oxidase

Ahmed S. Ibrahim, Sally Elshafey, Hassan Sellak, Khaled A. Hussein, Mohamed El-Sherbiny, Mohammed Abdelsaid, Nasser Rizk, Selina Beasley, Amany M. Tawfik, Sylvia B. Smith, Mohamed Al-Shabrawey

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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.

Original languageEnglish (US)
Pages (from-to)599-611
Number of pages13
JournalJournal of Lipid Research
Volume56
Issue number3
DOIs
StatePublished - Mar 1 2015

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12-Hydroxy-5,8,10,14-eicosatetraenoic Acid
NADPH Oxidase
Arachidonate 15-Lipoxygenase
Diabetic Retinopathy
Hyperglycemia
Metabolites
Endothelial cells
Permeability
Endothelial Cells
Angiography
Eicosanoids
Intraocular Injections
Streptozocin
Fluorescein
Arachidonic Acid
Macular Edema
Fluorescein Angiography
Albumins
Cell Movement
Catalytic Domain

Keywords

  • 12-and 15-HETEs
  • Bioactive lipids
  • Diabetic retinopathy
  • Lipoxygenase
  • Reduced nicotinamide adenine dinucleotide phosphate oxidase
  • Retinal inflammation
  • Retinal vascular leakage

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

Cite this

A lipidomic screen of hyperglycemia-treated hrecs links 12/15-lipoxygenase to microvascular dysfunction during diabetic retinopathy via NADPH oxidase. / Ibrahim, Ahmed S.; Elshafey, Sally; Sellak, Hassan; Hussein, Khaled A.; El-Sherbiny, Mohamed; Abdelsaid, Mohammed; Rizk, Nasser; Beasley, Selina; Tawfik, Amany M.; Smith, Sylvia B.; Al-Shabrawey, Mohamed.

In: Journal of Lipid Research, Vol. 56, No. 3, 01.03.2015, p. 599-611.

Research output: Contribution to journalArticle

Ibrahim, Ahmed S. ; Elshafey, Sally ; Sellak, Hassan ; Hussein, Khaled A. ; El-Sherbiny, Mohamed ; Abdelsaid, Mohammed ; Rizk, Nasser ; Beasley, Selina ; Tawfik, Amany M. ; Smith, Sylvia B. ; Al-Shabrawey, Mohamed. / A lipidomic screen of hyperglycemia-treated hrecs links 12/15-lipoxygenase to microvascular dysfunction during diabetic retinopathy via NADPH oxidase. In: Journal of Lipid Research. 2015 ; Vol. 56, No. 3. pp. 599-611.
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AU - Sellak, Hassan

AU - Hussein, Khaled A.

AU - El-Sherbiny, Mohamed

AU - Abdelsaid, Mohammed

AU - Rizk, Nasser

AU - Beasley, Selina

AU - Tawfik, Amany M.

AU - Smith, Sylvia B.

AU - Al-Shabrawey, Mohamed

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AB - 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.

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