IMMUNOCHEMICAL STUDIES OF LENS PIGMENTS

人的透镜核随着年龄的增长逐渐变黄， 白内障形成。这种核颜色，由快速测量仪精确测量， 光谱扫描测色法（FSSC）是一种新的颜色测量系统 强烈影响对比敏感度功能（CSF）。的能力 将颜色参数与视觉功能障碍直接相关是一个主要的 视觉研究的成就。然而，核颜色的起源是 还没有确定。三种机制可以产生黄色 色素在体外实验：光氧化，非酶先进 糖化和脂质过氧化。假设透镜色素 大多源于晚期糖基化，因为透镜生理学有利于 糖化机制然而，很难确定和量化 糖化色素或其它色素在人透镜中通过生物化学和 生物物理测量。这是因为色素发色团是 对酸水解不稳定且它们具有相似的分子光谱 特征（紫外-可见吸收和蓝色荧光）。免疫化学 测量可以提供替代方案。在这份报告中，主要 本发明的目的包括：（i）通过以下方法制备和表征颜料： 体外晚期糖基化、紫外光光氧化和脂质 过氧化;（2）分离荧光交联物质（着色 肽），并将它们作为抗原用于制备多克隆和 单克隆抗体;和（3）进行免疫化学测量 （Western blotting和ELISA），试图鉴定和定量 白内障手术后的透镜样本中的色素。这将使我们能够 确定导致透镜褐变的主要色素沉着机制 一种特殊类型的白内障这些知识可能有助于 减缓或阻止透镜布朗宁的药物方法的开发 并且可以为患者提供额外的非手术治疗选择， 白内障和年龄相关的视力丧失。

The human lens nucleus becomes progressively more yellow with age and with cataract formation. This nuclear color, measured accurately by the Fast Spectral Scanning Colorimetry (FSSC), a color measuring system, was found to affect strongly the contrast sensitivity function (CSF). The ability to correlate color parameters directly with visual dysfunction is a major accomplishment in vision research. However, the origin of nuclear color is still not definitely established. Three mechanisms can produce yellow pigments in in vitro experiments: photooxidation, nonenzymatic advanced glycation, and lipid peroxidation. It is hypothesized that lens pigments originate mostly from advanced glycation, since lens physiology favors the glycation mechanism. However, it is difficult to identify and quantify the glycation pigments or other pigments in the human lens by biochemical and biophysical measurements. This is because the pigment chromophores are labile to acid hydrolysis and they have similar molecular spectral features (UV-visible absorption and blue fluorescence). Immunochemical measurement may provide an alternative. In this proposal, the major objectives include: (i) to produce and characterize the pigments by in vitro advanced glycation, photooxidation of UV light, and lipid peroxidation; (2) to isolate fluorescent cross-linked species (pigmented peptides) and to use them as antigens in the preparation of polyclonal and monoclonal antibodies; and (3) to perform immunochemical measurements (Western blotting and ELISA) in an attempt to identify and quantify pigments in lens specimens from cataract surgery. This will permit us to define the predominant pigmentation mechanism that causes lens brunescence in a particular type of cataract. This knowledge may help in the development of a pharmaceutical means of slowing or stopping lens browning and could offer patients additional non-surgical options of treating cataract- and age-related vision loss.