COORDINATED REGULATION OF ANTIOXIDANT ENZYMES

这个项目将考察一个真正的双向 主要抗氧化酶之间存在协调调节 测定破坏细胞氧化剂动态平衡对生命的影响 原代人正常二倍体成纤维细胞的跨度。之前的研究由 其他人只是证明暴露在促氧化剂刺激下会增加 这些酶的活性。我们最近从基因上证明了 改变铜锌超氧化物抗氧化物酶的表达 歧化酶，调节其他抗氧化酶的表达，以及 氧化剂和非氧化剂相关蛋白。酶的变化很稳定 在不需要外源性促氧化剂刺激(高氧血症或 百草枯)。此外，一些变化似乎是协调一致的，因为两者 产生了增加和减少。我们检测到的法规 这些抗氧化酶之间可能是单向的，而 铜锌超氧化物歧化酶的表达影响细胞的表达 其他酶则不是这样。也就是说，如果是单向的，那么 过氧化锰或谷胱甘肽过氧化物酶的改变不会 影响铜锌超氧化物歧化酶的表达。这项规定， 然而，这些改变可以是双向或真正协调的 在锰中，超氧化物歧化酶和/或谷胱甘肽过氧化物酶将改变 铜锌超氧化物歧化酶的表达及相互表达。 这个项目的第一部分将决定一个真正的 抗氧化酶之间存在双向协调调节。 一种特定的抗氧化酶的活性将通过引入改变 正义或反义表达载体(使用连接的病毒启动子 到抗氧化酶的cdna)。这项规定将在两个地方进行研究 早传代和晚传代(衰老)成纤维细胞，以及 不朽的女儿线也决定了这一规定是否 在老化过程中或在永生时被破坏的。关联性 确定这些机构之间是否存在协调的监管 抗氧化酶，以及这一规定的存在如何将 各种看似无关的疾病，也进行了讨论。第二 该项目的一部分将检查改性抗氧化剂的效果 酶，再次通过引入正义或反义表达 载体，这些成纤维细胞的寿命。

This project will examine the possibility that a true bi-directional coordinated regulation exists between major anti-oxidant enzymes and determine the effect of disrupting cellular oxidant homeostasis on the life span of primary human normal diploid fibroblasts. Previous studies by others only demonstrate that exposure to pro-oxidant stimuli increases the activity of these enzymes. We recently demonstrated that genetically altering the expression of one anti-oxidant enzyme, copper-zinc superoxide dismutase, modifies expression of other anti-oxidant enzymes, as well as oxidant & non-oxidant associated proteins. Enzyme alterations are stable and occur without need of exogenous pro-oxidant stimuli (hyperoxia or paraquat). Also, some alterations appear to be coordinated in that both increases and decreases are produced. The regulation we have detected between these anti-oxidant enzymes may be uni-directional in that while copper-zinc superoxide dismutase expression influences expression of the other enzymes the reverse is not true. That is, if unidirectional then alterations in manganese superoxide or glutathione peroxidase would not influence copper-zinc superoxide dismutase expression. The regulation, however, could be bi-directional or truly coordinated in that alterations in manganese superoxide dismutase and/or glutathione peroxidase will alter expression of copperzinc superoxide dismutase and expression of each other. The first portion of this project will determine whether a true bi-directional coordinated regulation exists between anti-oxidant enzymes. Activity of a specific anti-oxidant enzyme will be altered by introduction of sense.or anti-sense expression vectors (using a viral promoter coupled to cDNA of an anti-oxidant enzyme). The regulation will be studied in both early passage and late passage (senescing) fibroblasts, as well as an immortalized daughter line to also determine if this regulation is disrupted during the aging process or upon immortalization. The relevance of determining whether a coordinated regulation exists amongst these anti-oxidant enzymes, and how the existence of this regulation could link a variety of seemingly unrelated diseases, is also discussed. The second portion of the project will examine the effect of modifying anti-oxidant enzymes, again through introduction of sense or anti-sense expression vectors, on the life span of these fibroblasts.