IDENTIFICATION OF FREE RADICAL RESISTANT GENES

衰老是功能丧失和 对压力的抵抗力或适应性。自由基已经被证明 在基本衰老过程中起重要作用。过表达 超氧化物歧化酶和过氧化氢酶已被证明可以增加平均 和果蝇的最大寿命（约30%）。饮食限制， 它可以防止氧化损伤，也可以延缓衰老过程， 在啮齿类动物中。自由基也与缩短寿命有关 以及与年龄相关的疾病，如阿尔茨海默氏病，帕金森氏病， 疾病和动脉粥样硬化。 为了研究介导自由基效应的分子机制， 自由基，我们已经建立了一个抗自由基的细胞系（PC 12-MR） 通过模拟慢性暴露于自由基的过程， 衰老由于PC 12-MR细胞对超氧化物具有抗性，因此表达 为了保护细胞免受自由基的损害，某些基因的某些部分必须改变。 可能的候选基因可以包括抗氧化酶（例如， 超氧化物歧化酶、过氧化氢酶和谷胱甘肽过氧化物酶）和细胞 死亡抑制剂（例如，Bcl-2和Bcl-XL）。我们的初步研究表明 还发现PC 12-MR细胞中cGMP水平升高。是 cGMP可能对氧化应激起保护作用。 由于哺乳动物细胞可能已经发展出多种系统来保护 在进化过程中抵抗环境和内源性氧化应激， 其他新基因的表达也可能发生改变， 在PC 12-MR细胞中，使它们抵抗自由基。审查 这些可能性，我们建议：l）检查是否表达 已知参与清除自由基和抑制 细胞凋亡在PC 12-MR细胞中发生改变，以及这些细胞凋亡是否被识别出来。 基因有助于保护PC 12-MR细胞免受自由基 损伤; 2）检测膜结合鸟苷酸环化酶/cGMP 对自由基发挥保护作用;以及3）识别 使PC 12-MR细胞抵抗自由基的其他新基因 使用差异显示技术的侮辱。的信息 这项研究产生的基因将有助于确定可以防止细胞凋亡的基因。 自由基导致的死亡。我的长期目标是了解 衰老的分子机制，特别是 保护细胞免受自由基损伤， 可开发用于延缓衰老过程，也可用于衰老- 相关疾病如阿尔茨海默病、帕金森病和 动脉粥样硬化

Aging is the consequence of the loss of functionality and the loss of resistance or adaptability to stress. Free radicals have been shown to play important roles in the basic aging process. Over-expression of superoxide dismutase and catalase has been shown to increase the mean and maximum life span (about 30%) of Drosophila. Dietary restriction, which protects against oxidative damage, also retards the aging process in rodents. Free radicals have also been implicated in life-shortening and age-associated disorders such as Alzheimer's disease, Parkinson's disease and atherosclerosis. To study the molecular mechanisms that mediate the effect of free radicals, we have developed a free radical-resistant cell line (PC12-MR) by mimicking the chronic exposure to free radicals in the process of aging. Since PC 12-MR cells are resistant to superoxide, the expression of some genes must be altered to protect cells from free radical damage. The possible candidate genes may include anti-oxidant enzymes (e.g., superoxide dismutase, catalase, and glutathione peroxidase) and cell death inhibitors (e.g., Bcl-2 and Bcl-XL). Our preliminary studies have also found that the level of cGMP in PC 12-MR cells is elevated. It is possible that cGMP may play a protective role against oxidative stress. Since mammalian cells may have developed multiple systems to protect against environmental and endogenous oxidative stress during evolution, it is possible that the expression of other novel genes is also altered in PC 12-MR cells to render them resistant to free radicals. To examine these possibilities, we propose: l) to examine whether the expression of genes known to be involved in scavenging free radicals and in inhibiting apoptosis are altered in PC 12-MR cells, and whether these identified genes contribute to the protection of PC 12-MR cells from free radical insult; 2) to examine whether membrane-bound guanylate cyclase/cGMP exerts a protective effect against free radicals; and 3) to identify other novel genes that render PC12-MR cells resistant to free radical insult employing the differential display technique. The information generated from this study will help identify genes that can prevent cell death caused by free radicals. My long-term goal is to understand the molecular mechanisms of aging and particularly the mechanisms that protect cells from free radical insult, so that therapeutic intervention may be developed to delay aging process and be applied as well to age- associated diseases such as Alzheimer disease, Parkinson's disease and atherosclerosis.