Age-related macular degeneration and retinal protein modification by 4-hydroxy-2-nonenal
Age-related macular degeneration and retinal protein modification by 4-hydroxy-2-nonenal
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DOI：
10.1167/iovs.06-1058
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发表时间：
2007-08-01
影响因子：
4.4
通讯作者：
Ferrington, Deborah A.
中科院分区：
文献类型：
--
作者：
Ethen, Cheryl M.;Reilly, Cavan;Ferrington, Deborah A.
目的。蛋白质、脂质和DNA的氧化损伤被认为是年龄相关性黄斑变性（AMD）的一种机制。视网膜特别容易受到脂质过氧化的影响，因为在充足的氧气存在下，多不饱和脂肪酸浓度很高，容易被氧化。脂质过氧化的毒性最大的产物之一，4-羟基-2-壬烯醛（HNE），可以修饰和灭活蛋白质。假设是4- hne修饰的蛋白会积累并作为amd进展阶段的标志。在AMD的四个进展阶段，在黄斑和周围区域都发现了含有HNE加合物的蛋白质。在检测hne内合蛋白之前，用二维（2-D）凝胶电泳对蛋白质进行溶解。利用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF/MS）对修饰蛋白进行鉴定。采用狭缝免疫印迹法比较HNE加合物的总含量。采用一维Western blot方法检测小鼠HNE解毒相关蛋白水平。19种蛋白被一致地修饰，无论AMD的阶段或视网膜区域。这些蛋白质有两个主要功能：能量产生和应激反应。在不同区域或分期之间，未观察到总hne内合蛋白的变化。观察到参与HNE解毒的蛋白质含量适度增加。一致修饰的蛋白质表明HNE氧化的首选蛋白质目标。HNE修饰的蛋白质在不同区域或阶段之间没有差异，这表明HNE解毒或去除受损蛋白质的途径是足够的。HNE修饰蛋白的一致水平表明HNE不是AMD的敏感视网膜生物标志物。
PURPOSE. Oxidative damage to proteins, lipids, and DNA has been suggested to be a mechanism for age- related macular degeneration (AMD). The retina is particularly susceptible to lipid peroxidation due to high concentrations of easily oxidized polyunsaturated fatty acids in the presence of abundant oxygen. One of the most toxic products of lipid peroxidation, 4-hydroxy-2-nonenal (HNE), can modify and inactivate proteins. The hypothesis was that 4-HNE-modified proteins would accumulate and serve as a marker for progressive stages of AMD.METHODS. Proteins containing HNE adducts were identified in both the macular and peripheral regions during four progressive stages of AMD. The proteins were resolved by two-dimensional (2-D) gel electrophoresis before detection of HNE-adducted proteins. Modified proteins were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS). The total content of HNE adducts was compared using a slot blot immunoassay. One-dimensional Western blot analysis was used to measure levels of proteins involved in HNE detoxification.RESULTS. Nineteen proteins that were consistently modified regardless of stage of AMD or retinal region were identified. These proteins are involved in two main functions: energy production and stress response. No change in total HNE-adducted protein was observed between regions or stages. Modest increases in content of proteins involved in HNE detoxification were observed.CONCLUSIONS. Consistently modified proteins indicate preferred protein targets for oxidation by HNE. HNE-modified proteins were not different between regions or stages, suggesting that pathways for detoxification of HNE or removal of damaged proteins are adequate. Consistent levels of HNE-modified proteins suggest that HNE is not a sensitive retinal biomarker for AMD.