Advanced Glycation End Product (AGE) Accumulation on Bruch's Membrane: Links to Age-Related RPE Dysfunction
Advanced Glycation End Product (AGE) Accumulation on Bruch's Membrane: Links to Age-Related RPE Dysfunction
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DOI：
10.1167/iovs.08-1724
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发表时间：
2009-01-01
影响因子：
4.4
通讯作者：
Stitt, Alan W.
中科院分区：
文献类型：
--
作者：
Glenn, Josephine V.;Mahaffy, Helen;Stitt, Alan W.
目的。晚期糖基化终产物（AGEs）在衰老过程中积累，并在死后的眼睛中观察到视网膜色素上皮（RPE）、布鲁氏膜和亚细胞沉积物（drusen）。年龄相关性黄斑变性（AMD）与年龄相关性视网膜功能障碍有关。在本研究中，我们评估了AGEs对RPE-Bruch膜界面的影响，以确定这些修饰如何导致与年龄相关的疾病。应用免疫组织化学方法评价Bruch膜上的AGEs。建立了具有临床意义的底物AGE体外模型来模拟Bruch膜老化。采用微阵列法研究ARPE-19在age修饰基底膜（AGE-BM）上生长1个月的反应，并采用定量RT-PCR验证。除了鉴定出与age相关的mRNA改变外，还研究了在age - bm上生长的ARPE-19的溶酶体酶活性和脂褐素积累。在临床标本的Bruch膜和脉络膜细胞外基质上观察到自体荧光和乙醇醛来源的AGEs。体外分析鉴定了暴露于AGE-BM的ARPE-19中一系列失调的mrna。暴露于AGE-BM后，观察到ARPE-19降解酶mRNA表达的改变。AGE- BM使ARPE-19组织蛋白酶- d活性显著降低（P < 0.05），脂褐素积累显著增加（P < 0.01）。AGEs影响ARPE-19 mRNA表达谱，并可能导致溶酶体酶降解能力降低和脂褐素积累增强。Bruch膜上AGEs的形成可能对年龄相关的外视网膜疾病有重要影响。（中国眼科杂志，2009;50:441-451）DOI:10.1167/iovs.08-1724
PURPOSE. Advanced glycation end products (AGEs) accumulate during aging and have been observed in postmortem eyes within the retinal pigment epithelium (RPE), Bruch's membrane, and subcellular deposits ( drusen). AGEs have been associated with age-related dysfunction of the RPE-in particular with development and progression to age-related macular degeneration (AMD). In the present study the impact of AGEs at the RPE-Bruch's membrane interface was evaluated, to establish how these modifications may contribute to age-related disease.METHODS. AGEs on Bruch's membrane were evaluated using immunohistochemistry. A clinically relevant in vitro model of substrate AGE accumulation was established to mimic Bruch's membrane ageing. Responses of ARPE-19 growing on AGE-modified basement membrane (AGE-BM) for 1 month were investigated by using a microarray approach and validated by quantitative (q)RT-PCR. In addition to identified AGE-related mRNA alterations, lysosomal enzyme activity and lipofuscin accumulation were also studied in ARPE-19 grown on AGE-BM.RESULTS. Autofluorescent and glycolaldehyde-derived AGEs were observed in clinical specimens on Bruch's membrane and choroidal extracellular matrix. In vitro analysis identified a range of dysregulated mRNAs in ARPE-19 exposed to AGE-BM. Altered ARPE-19 degradative enzyme mRNA expression was observed on exposure to AGE-BM. AGE- BM caused a significant reduction in cathepsin-D activity in ARPE-19 (P < 0.05) and an increase in lipofuscin accumulation (P < 0.01).CONCLUSIONS. AGEs influence ARPE-19 mRNA expression profiles and may contribute to reduced lysosomal enzyme degradative capacity and enhanced accumulation of lipofuscin. Formation of AGEs on Bruch's membrane may have important consequences for age-related outer retinal disease. (Invest Ophthalmol Vis Sci. 2009; 50: 441-451) DOI:10.1167/iovs.08-1724