Friedrichson T, Kalbach H L, Buck P, van Kuijk F J
Department of Chemistry and Biochemistry, Montana State University, Bozeman 59717, USA.
Curr Eye Res. 1995 Aug;14(8):693-701. doi: 10.3109/02713689508998497.
This study was undertaken to investigate the distribution of vitamin E in the macular and peripheral regions of the human retina, retinal pigment epithelium (RPE) and choroid as a function of age. High-performance liquid chromatography (HPLC) was used to measure alpha- and gamma-tocopherol levels quantitatively using tocol as an internal standard. In 57 out of 70 donor eyes (ages 9-104) the macular region was isolated and the tocopherols analyzed. The conventional brush method and a new vortex method were used to isolate the retinal pigment epithelium cells. Similar trends for the vitamin E levels (increase to the 5th decade, decrease after 7th decade) were found for the macular and peripheral retina and for the macular RPE. In the peripheral RPE a slight continuous increase with age was found. The vitamin E levels are higher in the RPE than in the retina, for both macular and peripheral regions. The amounts of vitamin E/mg protein are lower in the macular retina than in the peripheral retina, whereas in the RPE there is no difference in vitamin E content between macular and peripheral regions. A simple method based on a gentle vortex step was found to offer several advantages over the more generally used isolation of RPE cells based on brushing, and there was no difference in recovery of vitamin E in RPE cells when they were isolated by either isolation technique. It was also found that denominators, used to express the values of vitamin E in tissues should, be used with care since age dependent trends in parameters/denominators could be caused by trends in the denominators only.
本研究旨在调查维生素E在人类视网膜黄斑区和周边区域、视网膜色素上皮（RPE）及脉络膜中的分布情况，并探究其与年龄的关系。采用高效液相色谱法（HPLC），以生育三烯酚作为内标物，定量测定α-生育酚和γ-生育酚的水平。在70只供体眼中（年龄9 - 104岁），选取57只分离出黄斑区并分析生育酚含量。采用传统的刷取法和一种新的涡旋法分离视网膜色素上皮细胞。黄斑区和周边视网膜以及黄斑区视网膜色素上皮的维生素E水平呈现相似趋势（在第5个十年上升，在第7个十年后下降）。在周边视网膜色素上皮中，发现其随年龄略有持续上升。无论是黄斑区还是周边区域，视网膜色素上皮中的维生素E水平均高于视网膜。黄斑区视网膜中每毫克蛋白质的维生素E含量低于周边视网膜，而在视网膜色素上皮中，黄斑区和周边区域的维生素E含量并无差异。结果发现，基于温和涡旋步骤的简单方法相较于更常用的基于刷取的视网膜色素上皮细胞分离方法具有多种优势，并且无论采用哪种分离技术，视网膜色素上皮细胞中维生素E的回收率并无差异。研究还发现，用于表示组织中维生素E值的分母在使用时应谨慎，因为参数/分母中与年龄相关的趋势可能仅由分母的趋势导致。
