胡務亮2006-07-262018-07-112006-07-262018-07-112001http://ntur.lib.ntu.edu.tw//handle/246246/22862老化是一個奧祕的過程，由於對於老化的研究似乎 一時很難得到清楚的結論，因此有人認為早老症候 群的原因及機轉比較單純，應是研究老化的一種適 合對象。早老症(Hutchinson-Gilford 氏症)是一種 極為罕見的疾病，患者生長極度不良，嬰幼兒期就 出現一些老年人的特徵，如皮膚萎縮、禿頭、皮下 脂肪消失、頭皮血管突出、四肢瘦小及骨骼吸收 等。患者早期出現動脈硬化，通常十幾歲以前就死 於心臟血管疾病。早老症的原因目前還不清楚。而 大部分其他的早老症候群目前發現多半和基因修 補或複製的缺損有關，比如Werner syndrome 的WRN 基因是一個DNA helicase 的蛋白質。這些疾病表 現出相當明顯的紫外線敏感性、基因缺損及腫瘤等 現象。 過去關於早老症的研究發現，包括皮膚第四 型膠原纖維及彈性纖維增高、醣蛋白GP200 提高、 基因修補控制異常等，缺乏特異性。為了突破研究 上的困境，我們決定以differential display 試 圖去了解早老症的分子機轉。我們已有培養多株本 土性早老症皮膚纖維芽細胞，計劃以differential display 的技術，可以對早老症的病因做一些推 斷。我們先抽取正常皮膚纖維芽細胞以及早老症患 者皮膚纖維芽細胞中之RNA。所選取的細胞大約是 10 自12 代，此時兩種細胞的外型還沒有差異。早 老症的細胞大約只能分裂至20 代左右，而正常細 胞常常可以到30 代。我們將RNA 以非放射線法標 誌後，與micro-array 濾紙雜合。不過結果兩種細 胞的差異相當大，而且背景的雜訊也相當高。我們 經過多次努力仍然無法克服這個問題，另外實驗室 之設備也無法進行放射線元素之實驗，因此到目前 為止還沒有具體之成果產生。未來如果有機會，將 使用新一代的micro-array，由雜合至顯像分析完 全自動化，或可克服目前的困難。Aging is a mysterious process results from multiple factors, and studies of the causes of aging are often obscured by the complexity of this phenomenon. Therefore, it is suggested that progeria, which presents a less complicated etiology and phenotype may allow research to focus on a regulation site involved in development and aging. Hutchinson-Gilford progeria syndrome (progeria) is a rare disease characterized by selected features of premature aging. Presentations include a general failure of the child to thrive, and features of an aged appearance of the skin, alopecia, decreased subcutaneous fat, prominent scalp veins, thin limbs, and osteolysis. The formation of atherosclerotic plaques result in the death of the majority of progeria patients in the second decade of life. The etiology of progeria is still unknown. Some other progeroid syndromes have been found to be associated with defects in either DNA repair or replication. For example, the Werner syndrome WRN gene is a DNA helicase. These diseases usually demonstrate clear UV sensitivity or tumor formation. Previous studies on progeria showed many non-specific changes. In order to make a breakthrough the molecular basis of progeria, we plan to do differential display study. We have had cultured skin fibroblasts from domestic patients with progeria. We have extract RNA from skin fibroblasts from both normal controls and progeria patients. The passage numbers are from 10 to 12. Ordinarily the progeria cells survive not much beyond 20 generations, but normal cells usually survive more than 30 generations. We label the RNA with non-isotope method. The labeled probes were used to hybridize micro-array filter paper. The results showed that the differences between the two kinds of cells were quite large, and the background noise was high. We tried several times, but we still can not get readable result to further analysis. We hope in the future, modern micro-array techniques could be applied to this project, that both hybridization, signal development and reading will be automatic. Then we will have chance to overcome current obstacles.application/pdf1421885 bytesapplication/pdfzh-TW國立臺灣大學醫學院小兒科早老症Hutchinson-Gilford 氏症differential displayProgeriaHutchinson-Gilford syndromedifferential displayreporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/22862/1/892320B002229.pdf