The expression of advanced glycation endproduct receptors in rpe cells associated with basal deposits in human maculas
The expression of advanced glycation endproduct receptors in rpe cells associated with basal deposits in human maculas
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DOI：
10.1016/j.exer.2005.10.005
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发表时间：
2006-05-01
影响因子：
3.4
通讯作者：
Handa, James T.
中科院分区：
文献类型：
--
作者：
Yamada, Yuko;Ishibashi, Kazuko;Handa, James T.
布鲁赫膜内的基底沉积物与衰老和年龄相关性黄斑变性（AMD）相关，尽管引起其形成的因素尚不完全清楚。晚期糖基化终产物（AGEs）随着年龄的增长在布鲁赫膜中积累，包括基底沉积物和玻璃疣。AGE改变细胞表型的一种机制是通过AGE受体。本研究的目的是免疫定位和定量的表达AGE受体的RPE细胞与基底存款或正常布鲁赫的膜，从人类黄斑显微解剖。将来自14名老年对照供体和5名患有非新生血管性AMD的供体的死后眼睛冷冻保存。激光捕获显微切割与正常Bruch膜或基底沉积物相关的RPE细胞。提取RNA并用于RT-qPCR，以定量AGEs、AGE R1、AGE R2和AGE R3的表达。还对这些受体进行了链霉亲和素碱性磷酸酶免疫组化，并从14名正常和9名AMD供体中漂白切片。RT-qPCR显示，与附着在形态正常的布鲁赫膜上的细胞相比，覆盖基底沉积物的RPE细胞中的AGEs、AGE R1和AGE R3显著上调。免疫组化分析显示，RPE细胞的弥漫性，染色浅，强烈的脉络膜毛细血管染色在正常的布鲁赫膜的区域。在基底沉积物的区域中，与正常布鲁赫膜区域相比，RPE具有更强的SAGE和AGER 1染色。这些结果表明，AGE受体可能影响老化和AMD过程中基底沉积的形成。(c)2005爱思唯尔有限公司保留所有权利。
Basal deposits within Bruch's membrane are associated with aging and age-related macular degeneration (AMD) although the factors causing their formation are incompletely understood. Advanced glycation endproducts (AGEs) accumulate in Bruch's membrane including basal deposits and drusen with aging. One mechanism by which AGEs alter a cell's phenotype is via AGE receptors. The purpose of this study was to immunolocalize and quantify the expression of AGE receptors by RPE cells associated with basal deposits or normal Bruch's membrane that were microdissected from human maculas. Postmortem eyes from 14 aged control donors and five donors with non-neovascular AMD were cryopreserved. RPE cells associated with normal Bruch's membrane or basal deposits were laser capture microdissected. The RNA was extracted and used for RT-qPCR to quantify the expression of RAGE, AGE R1, AGE R2, and AGE R3. Streptavidin alkaline phosphatase immunohistochemistry for these receptors was also performed and sections were bleached from 14 normal and nine AMD donors. RT-qPCR showed significant upregulation of RAGE, AGE R1, and AGE R3 in RPE cells overlying basal deposits compared to cells attached to morphologically normal Bruch's membrane. Immunohistochemical analysis for RAGE, AGER1, R2, and R3 showed diffuse, light staining of RPE cells and strong choriocapillaris staining in areas of normal Bruch's membrane. In areas of basal deposits, the RPE had more intense staining for RAGE and AGER1 compared to regions of normal Bruch's membrane. These results suggest that AGE receptors could influence the formation of basal deposits during aging and AMD. (c) 2005 Elsevier Ltd. All rights reserved.