OCULAR LENS STUDIES

本申请中描述的工作旨在探索这一前提 晶状体上皮细胞氧化还原状态可能调节基因表达。 有许多基因的产物调节着 其他基因。有人提出，这些调控基因中的一些，如 FOS、JUN和NF-kappaB本身受晶状体系统中氧化还原的控制。 这些调控基因之所以被选中，是因为它们分布广泛， 受氧化还原的影响，其产物的结合部位 据报道与某些晶体蛋白基因有关。实验性的 本申请中描述的工作将阐明 氧化应激改变这些控制基因的表达，并 随后影响报告基因和晶状体基因的表达。 对正常细胞基因调控被破坏的论证 氧化应激将为这一机制提供一个新的基本方面 氧化应激可导致白内障，并更加强调 发生在上皮细胞中的事件。具体地说，计划 静息状态下晶状体中FOS、JUN和核因子-kappaB的表达 以及在正常和氧化应激条件下分裂细胞系。这个 Fos、Jun和NF-kappaB对报告基因和某些晶状体的调控 含有AP-1增强子和核因子-kappaB结合位点的基因将 检查过了。细胞氧化还原设定点将基于GSH/GSSG确定， NADH/NAD和NADPH/NADP。消除某些方面的药物的效果 氧化应激，如AL-3823A，去铁胺和TEMPO也将 调查过了。据报道，初步观察表明 这一方法的可行性。

The work described in this application is designed to explore the premise that the lens epithelial cell redox status may regulate gene expression. There are a number of genes whose products regulate the expression of other genes. It is proposed that some of these regulatory genes such as fos, jun and NF-kappaB are themselves controlled by redox in lens systems. These regulatory genes have been chosen since they are widely distributed, are effected by redox and binding sites for their products have been reported to be associated with certain crystallin genes. The experimental work described in this application will elucidate the extent to which oxidative stress alters the expression of these control genes and subsequently effects the expression of a reporter gene and of lens genes. The demonstration of the disruption of normal cellular gene regulation by oxidative stress would provide a new fundamental aspect to the mechanism by which oxidative stress can cause cataract and place greater emphasis on events occurring in the epithelial cells. Specifically, it is planned to determine the expression of fos, jun and NF-kappaB in the lens in resting and dividing cell lines under normal and oxidative stress conditions. The regulation by fos, jun and NF-kappaB of a reporter gene and certain lens genes containing AP-1 enhancer sites and NF-kappaB binding sites will be examined. Cellular redox set points will be determined based on GSH/GSSG, NADH/NAD and NADPH/NADP. The effect of agents which eliminate aspects of oxidative stress such as AL-3823A, desferoxamine and Tempo will also be investigated. Preliminary observations are reported indicating the feasibility of this approach.