12/15-Lipoxygenase-Derived Lipid Metabolites Induce Retinal Endothelial Cell Barrier Dysfunction: Contribution of NADPH Oxidase

Amira Othman, Saif Ahmad, Sylvia Megyerdi, Rene Mussell, Karishma Choksi, Krishna Rao Maddipati, Ahmed Abdelrazik Elmarakby, Nasser Rizk, Mohamed Al-Shabrawey

Research output: Contribution to journalArticle

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Abstract

The purpose of the current study was to evaluate the effect of 12/15- lipoxygenase (12/15-LOX) metabolites on retinal endothelial cell (REC) barrier function. FITC-dextran flux across the REC monolayers and electrical cell-substrate impedance sensing (ECIS) were used to evaluate the effect of 12- and 15-hydroxyeicosatetreanoic acids (HETE) on REC permeability and transcellular electrical resistance (TER). Effect of 12- or 15-HETE on the levels of zonula occludens protein 1 (ZO-1), reactive oxygen species (ROS), NOX2, pVEGF-R2 and pSHP1 was examined in the presence or absence of inhibitors of NADPH oxidase. In vivo studies were performed using Ins2Akita mice treated with or without the 12/15-LOX inhibitor baicalein. Levels of HETE and inflammatory mediators were examined by LC/MS and Multiplex Immunoassay respectively. ROS generation and NOX2 expression were also measured in mice retinas. 12- and 15- HETE significantly increased permeability and reduced TER and ZO-1expression in REC. VEGF-R2 inhibitor reduced the permeability effect of 12-HETE. Treatment of REC with HETE also increased ROS generation and expression of NOX2 and pVEGF-R2 and decreased pSHP1 expression. Treatment of diabetic mice with baicalein significantly decreased retinal HETE, ICAM-1, VCAM-1, IL-6, ROS generation, and NOX2 expression. Baicalein also reduced pVEGF-R2 while restored pSHP1 levels in diabetic retina. Our findings suggest that 12/15-LOX contributes to vascular hyperpermeability during DR via NADPH oxidase dependent mechanism which involves suppression of protein tyrosine phosphatase and activation of VEGF-R2 signal pathway.

Original languageEnglish (US)
Article numbere57254
JournalPloS one
Volume8
Issue number2
DOIs
StatePublished - Feb 20 2013

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NADPH Oxidase
Endothelial cells
Metabolites
lipoxygenase
endothelial cells
Endothelial Cells
metabolites
Lipids
Acids
acids
lipids
reactive oxygen species
Reactive Oxygen Species
Electric Impedance
Lipoxygenase Inhibitors
Permeability
permeability
Acoustic impedance
electrical resistance
retina

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

12/15-Lipoxygenase-Derived Lipid Metabolites Induce Retinal Endothelial Cell Barrier Dysfunction : Contribution of NADPH Oxidase. / Othman, Amira; Ahmad, Saif; Megyerdi, Sylvia; Mussell, Rene; Choksi, Karishma; Maddipati, Krishna Rao; Elmarakby, Ahmed Abdelrazik; Rizk, Nasser; Al-Shabrawey, Mohamed.

In: PloS one, Vol. 8, No. 2, e57254, 20.02.2013.

Research output: Contribution to journalArticle

Othman, Amira ; Ahmad, Saif ; Megyerdi, Sylvia ; Mussell, Rene ; Choksi, Karishma ; Maddipati, Krishna Rao ; Elmarakby, Ahmed Abdelrazik ; Rizk, Nasser ; Al-Shabrawey, Mohamed. / 12/15-Lipoxygenase-Derived Lipid Metabolites Induce Retinal Endothelial Cell Barrier Dysfunction : Contribution of NADPH Oxidase. In: PloS one. 2013 ; Vol. 8, No. 2.
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