Eldred G E, Katz M L
University of Missouri School of Medicine, Department of Ophthalmology, Columbia 65212.
Free Radic Biol Med. 1989;7(2):157-63. doi: 10.1016/0891-5849(89)90007-5.
The fluorescent molecules of cellular age pigment granules (lipofuscin) are commonly thought to be end products of membrane lipid autoxidation. Lipofuscin fluorophores of the retinal pigment epithelium (RPE) appear to be derived from photoreceptor outer segment membranes. Experiments were therefore conducted to determine whether the in vitro oxidation of retinal homogenates would generate fluorophores similar to the naturally occurring lipofuscin fluorophores of the RPE. Neural retina and RPE-choroid homogenates from young (2-3 month old) albino rats were subjected to an iron-ascorbate-air pro-oxidant reaction medium, and compared to unoxidized control samples from young age-matched animals as well as senescent (24 month old) rats. In addition, neural retina and RPE-choroid homogenates from 3 month old albino rats were subjected to a 100% oxygen atmosphere to test whether the fluorescent products of autoxidation differ substantially from those generated in the pro-oxidant medium. The chloroform-soluble fluorophores of chloroform-methanol sample extracts were analyzed by corrected fluorescence spectroscopy and thin-layer chromatography (TLC). In vitro pro-oxidation of both the neural retina and the RPE from young rats produced blue-emitting fluorophores which differed from the orange- and yellow-emitting fluorophores extracted from the RPE of senescent rats. Corrected fluorescence spectroscopy of aged tissue extracts revealed vitamin A-related fluorescence (330 nm excitation maximum; 515 nm emission maximum) and a spectrally resolvable age-related fluorescence (420 nm excitation maximum; 600 nm emission maximum). Only the vitamin A-related fluorescence could be measured in the control of young samples.(ABSTRACT TRUNCATED AT 250 WORDS)
细胞衰老色素颗粒(脂褐素)的荧光分子通常被认为是膜脂质自动氧化的终产物。视网膜色素上皮(RPE)的脂褐素荧光团似乎源自光感受器外段膜。因此进行了实验,以确定视网膜匀浆的体外氧化是否会产生与RPE天然存在的脂褐素荧光团相似的荧光团。将幼年(2 - 3个月大)白化大鼠的神经视网膜和RPE - 脉络膜匀浆置于铁 - 抗坏血酸 - 空气促氧化反应介质中,并与来自年龄匹配的幼年动物以及衰老(24个月大)大鼠的未氧化对照样品进行比较。此外,将3个月大的白化大鼠的神经视网膜和RPE - 脉络膜匀浆置于100%氧气气氛中,以测试自动氧化的荧光产物是否与促氧化介质中产生的荧光产物有很大不同。通过校正荧光光谱法和薄层色谱法(TLC)分析氯仿 - 甲醇样品提取物中氯仿可溶的荧光团。幼年大鼠的神经视网膜和RPE的体外促氧化产生了发出蓝光的荧光团,这与从衰老大鼠的RPE中提取的发出橙色和黄色光的荧光团不同。衰老组织提取物的校正荧光光谱显示与维生素A相关的荧光(最大激发波长330nm;最大发射波长515nm)和在光谱上可分辨的与年龄相关的荧光(最大激发波长420nm;最大发射波长600nm)。在幼年样品的对照中只能测量到与维生素A相关的荧光。(摘要截短于250字)