The Role of Cytochrome P450 Epoxygenases in Retinal Angiogenesis
The Role of Cytochrome P450 Epoxygenases in Retinal Angiogenesis
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DOI：
10.1167/iovs.14-14216
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发表时间：
2014-07-01
影响因子：
4.4
通讯作者：
Penn, John S.
中科院分区：
文献类型：
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作者：
Capozzi, Megan E.;McCollum, Gary W.;Penn, John S.
目的。本研究旨在探讨细胞色素P450环氧合酶及其产物环氧二十碳三烯酸(EETs)在低氧诱导的血管内皮生长因子生成和病理性视网膜血管生成中的作用。方法：将人视网膜星形胶质细胞、Muller细胞和视网膜微血管内皮细胞暴露于低氧环境中，用RT-PCR方法检测细胞色素P450环氧合酶C的相对表达。用酶联免疫吸附试验检测低氧条件下和常规CYP抑制剂SKF-525A处理后星形胶质细胞和Muller细胞的血管内皮细胞生长因子的表达。人视网膜微血管内皮细胞用CYP产物11，12-环氧二十碳三烯酸[EET]或SKF-525A在有或没有血管内皮生长因子的情况下处理。检测HRMEC的增殖和管状形成。以新生大鼠为模型，建立氧源性视网膜病变模型。逆转录-聚合酶链式反应(RT-PCR)检测视网膜组织中细胞色素P450 2C11和细胞色素P3 2 3的表达。OIR大鼠玻璃体内注射SKF-525A，对视网膜前新生血管(NV)进行定量。结果：人视网膜星形胶质细胞在低氧条件下可显著诱导细胞色素P450 2C8和细胞色素P450 2C9的表达。当用SKF-525A处理时，星形胶质细胞，而不是Muller细胞，表现出低氧诱导的VEGF产生减少。11，12-EET诱导HRMEC增殖和管状形成，SKF-525A抑制血管内皮生长因子诱导的细胞增殖。氧诱导的视网膜病变诱导了细胞色素P4 2 2 3的表达，但对细胞色素P2 2 C11的表达无影响。SKF-525A抑制OIR大鼠视网膜新生血管和降低视网膜血管内皮细胞生长因子水平。结论星形胶质细胞缺氧刺激后，CYP来源的11，12-EET可能在视网膜发挥促血管生成的生物学功能。抑制CYP可能是一种治疗视网膜新生血管的合理方法，因为它可以减少血管内皮细胞中血管内皮生长因子的产生和诱导的血管生成反应。
PURPOSE. The purpose of this study was to investigate the role(s) of cytochrome P450 epoxygenases (CYPs) and their products, the epoxyeicosatrienoic acids (EETs), in hypoxia-induced VEGF production and pathologic retinal angiogenesis.METHODS. Human retinal astrocytes, Muller cells, and retinal microvascular endothelial cells (HRMEC) were exposed to hypoxia, and relative CYP2C expression was measured by RT-PCR. Astrocyte and Muller cell VEGF production was measured by ELISA after exposure to hypoxia and treatment with the general CYP inhibitor, SKF-525a. Human retinal microvascular endothelial cells were treated with the CYP product, 11,12-epoxyeicosatrienoic acid [EET], or SKF-525a in the presence or absence of VEGF. Proliferation of HRMEC and tube formation were assayed. Oxygen-induced retinopathy (OIR) was induced in newborn rats. Retinal CYP2C11 and CYP2C23 expression were measured by RT-PCR. The OIR rats received SKF-525a by intravitreal injection and preretinal neovascularization (NV) was quantified. Retinal VEGF protein levels were measured by ELISA.RESULTS. Human retinal astrocytes were the only cells to exhibit significant induction of CYP2C8 and CYP2C9 mRNA expression by hypoxia. Astrocytes, but not Muller cells, exhibited reduced hypoxia-induced VEGF production when treated with SKF-525a. 11,12-EET induced HRMEC proliferation and tube formation, and SKF-525a inhibited VEGF-induced proliferation. Oxygen-induced retinopathy induced expression of CYP2C23, but had no effect on CYP2C11. SKF-525a inhibited retinal NV and reduced retinal VEGF levels in OIR rats.CONCLUSIONS. The CYP-derived 11,12-EET may exhibit a proangiogenic biological function in the retina following stimulation by hypoxia in astrocytes. Inhibition of CYP may provide a rational therapy against retinal NV, because it can reduce VEGF production and VEGF-induced angiogenic responses in endothelial cells.