- 學位論文
人類纖維母細胞於不同生長因子中所引發之類似老化之表徵
The effect of growth factor on the induction of senescence-like phenotype of human dermal fibroblast
摘要
前言: 隨著全球人口老化的趨勢，如何抗老化就變成是一個非常重要的課題；被譽為青春之鑰的生長因子就變成大家爭相研究的對象。但至今尚未有文獻指出，將人類纖維母細胞長期連續的施予生長因子對於細胞老化上的影響。因此，本研究的主要目標為觀察不同生長因子長期培養對於細胞老化的影響。 材料與方法：表皮生長因子和人類纖維母細胞生長因子作為實驗的生長因子。利用人類真皮纖維母細胞作為實驗細胞，培養於不同的生長因子中，建立細胞長期之生長曲線與 SA-β-galactosidase酵素活性等老化相關指標，最後進行和老化相關的基因分析。 結果：表皮生長因子(EGF)的實驗組中，在短天期由生長曲線觀察得知細胞數量沒有明顯差異代表細胞增生能力沒有增加，但長天期後生長曲線、細胞型態、Senescence-associated ß-galactosidase活性和老化相關基因表現均產生明顯類似於老化的型態。纖維母細胞生長因子(bFGF)實驗組中，在經過較長天期後由生長曲線觀察得知細胞數量有明顯差異代表胞增生能力增加，在繼續培養下去長曲線、細胞型態、Senescence-associated ß-galactosidase活性和老化相關基因表現均產生明顯類似於老化的型態。 結論：加表皮生長因子(EGF)的實驗組中，表皮生長因子(EGF)不利於人類纖維母細胞生長，就像是一種壓力提早加速細胞老化與凋亡。另外在纖維母細胞生長因子(bFGF)實驗組中，在實驗早期會增加細胞proliferation，但正常體細胞端粒有一定的長度，纖維母細胞生長因子(bFGF)會使細胞複製的次數比較多而提早造成複製性老化。
並列摘要
Introduction: The aging is a very important issue for human with the global trend of an aging population. Therefore, the youth of the key growth factor was become study object nowadays .Hence, the main objective was developing long-term effects of different growth factors for cell aging in this study. Materials and methods: Human epidermal growth factor and fibroblast growth factor as a test of the growth factor. Primary skin dermal fibroblasts were cultured on two growth factor to establish senescence correlation index, such as growth curves and SA-β-galactosidase activity. Finally, the senescence associated gene analysis were carried out. Results:Epidermal growth factor (EGF) was observed no significant difference from the P.D curve in the short-term period. In the long-term period, the P.D curve, cell morphology, Senescence-associated ß-galactosidase activity and aging-related gene expression were significantly similar to the aging of the type produced. In the experimental group of plus Fibroblast growth factor (bFGF), the P.D curves were observed after a long -term period that are significant differences in the number of cells representative of the ability to increase cell proliferation. And continuing the curve, the cell morphology, Senescence-associated ß-galactosidase activity and aging-related gene expression were significantly similar to the aging of the type produced. Conclusion: According to the result of the sample containing Epidermal growth factor, Epidermal growth factor may not be able to enhance the growth of Fibroblast. In the Epidermal growth factor containing specimen, Epidermal growth factor may become a stress which induces the senescence and apoptosis of cell. In addition, the result of the sample containing Fibroblast growth factor shows increase of cell proliferation in early stage. (Generally, normal telomere has fixed length.) Fibroblast growth factor could increase the proliferation times of cell and therefore approach replicative senescence earlier than usual.