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通过二维凝胶电泳蛋白质图谱检测发现,小梁网中存在保护氧化损伤的酶,且在生命的各个年龄段均有发现。

Enzymes protective of oxidative damage present in all decades of life in the trabecular meshwork, as detected by two-dimensional gel electrophoresis protein maps.

作者信息

Russell P, Johnson D H

机构信息

National Institutes of Health, Bethesda, Maryland 20892-2735, USA.

出版信息

J Glaucoma. 1996 Oct;5(5):317-24.

PMID:8897231
Abstract

Our purpose was to determine whether there are age-related differences in the presence of enzymes protective against oxidative damage in normal human trabecular meshwork (TM). The study was also done to determine whether there are significant changes in proteins related to age, gender, or position around the circumference of the meshwork. Human TM was dissected from both eyes of 16 normal donors ranging in age from 9 to 91 years. Proteins were mapped with two-dimensional (2D) gel electrophoresis. The microanalytical method used allowed analysis of samples as small as one fourth of a single meshwork. Selected proteins were identified by Western blot. Superoxide dismutase (SOD) and glutathione reductase were present in all ages on the 2D maps. Age-related changes in the TM were not apparent, although slight differences may exist between juvenile and adult TM, since differences in the relative intensities of some polypeptides were evident. In any single quadrant of the TM, > or = 160 protein spots could be counted visually. The 2D maps of the various quadrants were very similar, and the protein patterns were analogous between fellow eyes. Distinct differences were noted between maps from TM and adjacent cornea and sclera. Other protein spots identified included actin, albumin, transferrin, and vimentin. The TM samples had a mean cellular protein content of 66.3 +/- 15.4 micrograms.

摘要

我们的目的是确定在正常人小梁网(TM)中,是否存在与年龄相关的抗氧化损伤保护酶差异。本研究还旨在确定与年龄、性别或小梁网圆周位置相关的蛋白质是否存在显著变化。从16名年龄在9至91岁之间的正常供体的双眼摘取人小梁网。蛋白质通过二维(2D)凝胶电泳进行图谱分析。所采用的微量分析方法能够分析小至单个小梁网四分之一大小的样本。通过蛋白质印迹法鉴定选定的蛋白质。在二维图谱上,超氧化物歧化酶(SOD)和谷胱甘肽还原酶在所有年龄段均有存在。小梁网中与年龄相关的变化并不明显,尽管青少年和成人小梁网之间可能存在细微差异,因为一些多肽的相对强度存在明显差异。在小梁网的任何一个象限中,肉眼可计数的蛋白质斑点≥160个。各个象限的二维图谱非常相似,且双眼之间的蛋白质模式类似。在小梁网与相邻角膜和巩膜的图谱之间发现了明显差异。鉴定出的其他蛋白质斑点包括肌动蛋白、白蛋白、转铁蛋白和波形蛋白。小梁网样本的细胞蛋白质平均含量为66.3±15.4微克。

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