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原发性开角型青光眼房水中氧化应激蛋白质组水平。

Aqueous humor oxidative stress proteomic levels in primary open angle glaucoma.

机构信息

University of Genova, Eye Clinic, Department of Neurosciences, Ophthalmology and Genetics, University of Genoa, Genoa, Italy.

出版信息

Exp Eye Res. 2012 Oct;103:55-62. doi: 10.1016/j.exer.2012.07.011. Epub 2012 Aug 21.

Abstract

The purpose of this work was to investigate the expression of glutamine synthase (GS), nitric oxide synthase (NOS) superoxide dismutase (SOD) and glutathione transferase (GST) in the aqueous humor of patients with primary open angle glaucoma and controls. Aqueous humor proteome was analyzed by antibody microarray. The expression of tested proteins was detected by protein Cy3/Cy5 labeling, column purification and hybridization on antibody-spotted glass microarray. Fluorescent signals were detected by fluorescence laser scanning. Aqueous humor levels of SOD as well as of GST were significantly lower (2.0- and 2.2-fold, p < 0.01) among patients than controls; both NOS and GS expression were significantly higher (2.2- and 2.6 fold, p < 0.01) among patients than controls. Our data showed substantial differences of GS, NOS2, SOD and GST aqueous humor levels between glaucomatous patients and controls as measured by antibody microarray technology. The overproduction of NO through inducible NOS can form toxic products and change the metabolic conditions of the TM. The GS over-expression might be related to neuronal injury or to the potential role of glutamate as a modulator in the ciliary body signaling. The reduced expression of the antioxidant enzymes SOD and GST could aggravate the unbalance between both oxygen- and nitrogen-derived free radicals production and detoxification. Based on our results, GS, NOS2, SOD and GST as measured by antibody microarray technology may be useful oxidative markers in aqueous humor of glaucomatous patients.

摘要

本研究旨在探讨谷氨酰胺合成酶(GS)、一氧化氮合酶(NOS)、超氧化物歧化酶(SOD)和谷胱甘肽转移酶(GST)在原发性开角型青光眼患者和对照组眼水中的表达。采用抗体微阵列分析房水蛋白质组。用蛋白 Cy3/Cy5 标记、柱纯化和抗体点样玻璃微阵列杂交检测所测试蛋白的表达。用荧光激光扫描检测荧光信号。与对照组相比,患者眼水中 SOD 和 GST 的水平显著降低(2.0 倍和 2.2 倍,p < 0.01);NOS 和 GS 的表达均显著升高(2.2 倍和 2.6 倍,p < 0.01)。通过抗体微阵列技术,我们的数据显示,青光眼患者与对照组之间 GS、NOS2、SOD 和 GST 眼水中的水平存在显著差异。诱导型 NOS 产生的过量 NO 可形成有毒产物并改变 TM 的代谢条件。GS 的过表达可能与神经元损伤有关,或与谷氨酸作为睫状体信号转导中的调节剂的潜在作用有关。抗氧化酶 SOD 和 GST 的表达减少会加重氧和氮衍生自由基产生与解毒之间的失衡。基于我们的结果,通过抗体微阵列技术测量的 GS、NOS2、SOD 和 GST 可能是青光眼患者眼水中有用的氧化标志物。

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