A lipidomic screen of hyperglycemia-treated HRECs links 12/15-Lipoxygenase to microvascular dysfunction during diabetic retinopathy via NADPH oxidase
A lipidomic screen of hyperglycemia-treated HRECs links 12/15-Lipoxygenase to microvascular dysfunction during diabetic retinopathy via NADPH oxidase
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DOI：
10.1194/jlr.m056069
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发表时间：
2015-03-01
影响因子：
6.5
通讯作者：
Al-Shabrawey, Mohamed
中科院分区：
文献类型：
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作者：
Ibrahim, Ahmed S.;Elshafey, Sally;Al-Shabrawey, Mohamed
视网膜通透性过高和随后的黄斑水肿是早期糖尿病视网膜病变（DR）的主要特征。在这里，我们研究了生物活性脂质代谢物，特别是 12/15-脂氧合酶 (LOX) 衍生的代谢物在此过程中的作用。人视网膜内皮细胞 (HREC) 的 LC/MS 脂质组学筛选表明，15-HETE 是高葡萄糖 (30 mM) 处理后唯一显着增加的代谢物（2.4 +/- 0.4 倍，P = 0.0004）。在花生四烯酸存在下，12/15-LOX 产生的额外类二十烷酸（包括 12- 和 11-HETE）显着增加。荧光素血管造影和视网膜白蛋白渗漏显示，与糖尿病 WT 小鼠相比，链脲佐菌素诱导的缺乏 12/15-LOX 的糖尿病小鼠的视网膜高渗透性显着降低。我们之前的研究证明了 NADPH 氧化酶在介导 12- 和 15-HETE 的通透性作用中的潜在作用，因此我们测试了眼内注射 12-HETE 对缺乏 NADPH 氧化酶 (NOX2) 催化亚基的小鼠的影响。与WT小鼠相比，NOX2(-/-)小鼠中12-HETE的通透性效应显着降低。体外实验还表明，15-HETE 以 NOX 依赖性方式诱导 HREC 迁移和管形成。综上所述，我们的数据表明 12/15-LOX 通过 NOX 依赖性机制参与 DR。
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.