Nitrite-modified extracellular matrix proteins deleteriously affect retinal pigment epithelial cell function and viability: a comparison study with nonenzymatic glycation mechanisms.
Nitrite-modified extracellular matrix proteins deleteriously affect retinal pigment epithelial cell function and viability: a comparison study with nonenzymatic glycation mechanisms.
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亚硝酸盐修饰的细胞外基质蛋白有害地影响视网膜色素上皮细胞的功能和活力：与非酶糖化机制的比较研究。
DOI：
10.1080/02713680590968259
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Gaillard,ElizabethR
中科院分区：
文献类型：
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作者：
Wang,Zhen;Paik,DavidC;DelPriore,LucianV;Burch,RobertL;Gaillard,ElizabethR
目的：细胞外基质（Extracellular matrix， ECM）在细胞功能调控中起重要作用。老化过程可能涉及到ECM蛋白的化学修饰，这可能导致Bruch膜的老化和年龄相关性黄斑变性（AMD）的发病机制。本研究旨在探讨亚硝酸盐修饰基底膜样蛋白对RPE细胞行为的影响，并以此作为年龄相关性眼病中Bruch膜老化的模型。作为比较，我们还研究了视网膜色素上皮细胞在乙醇醛修饰基质（GMM）上的行为。方法：生长因子还原Matrigel分别与亚硝酸盐和乙醇醛反应1周和12小时。小牛RPE细胞被镀在修饰的基质上，并以几种方式进行检测。测定附着率、增殖率、细胞凋亡和坏死。还监测了细胞形态和细胞对a2e介导的损伤的易感性。结果：亚硝酸盐修饰基质（nmm）对细胞附着的抑制率为65%，对细胞增殖的抑制率为33.7%，而GMM分别为69.6%和21.7%。高密度GMM对细胞增殖抑制不显著（3.47%），而NMM对细胞增殖抑制显著（20.9%）。NMM诱导细胞凋亡和坏死，而GMM仅诱导细胞凋亡。两种修饰均抑制RPE分化。两种基质下的RPE细胞都更容易受到A2E介导的蓝光损伤，但NMM的损伤更大。结论：NMM对RPE细胞功能和活力具有明显的损害作用，其损害作用与GMM相似。这些研究可能与AMD进展过程中观察到的RPE功能障碍有关。
Purpose: Extracellular matrix (ECM) plays an important role in the regulation of cell function. The aging process may involve chemical modifications to ECM proteins, which may contribute to the aging of the Bruch membrane and pathogenesis of age-related macular degeneration (AMD). The purpose of this study is to investigate nitrite modification of basement membrane-like proteins on RPE cell behavior as a model for the aging of the Bruch membrane in age-related eye diseases. As a comparison, retinal pigment epithelium (RPE) cell behavior on glycolaldehyde-modified matrices (GMM) was also studied.Methods: Growth factor reduced Matrigel was reacted with nitrite or glycolaldehyde for 1 week or 12 hr, respectively. Calf RPE cells were plated on the modified matrices and examined in several ways. Attachment rates, proliferation rates, apoptosis, and necrosis were determined. Cell morphology and cell susceptibility to A2E-mediated damage was also monitored.Results: Nitrite-modified matrices (NMMs) inhibited cell attachment by 65% and proliferation by 33.7% compared to 69.6% and 21.7%, respectively, by GMM. Proliferation inhibition was not significant when cells were plated at high density on GMM (3.47%) but significant on NMM (20.9%). NMM induced cell apoptosis and necrosis, but GMM induced cell apoptosis only. Both modifications inhibited RPE differentiation. RPE cells on both matrices were more susceptible to blue light mediated damage by A2E, but damage was greater on NMM.Conclusions: NMM has significant damaging effects on RPE cell function and viability that is similar to the damaging effects of GMM. These studies may have relevance to the RPE dysfunction observed during the progression of AMD.