- 學位論文
正常成纖維細胞株與蟹足腫成纖維細胞株內乙型轉型生長因子訊息傳遞之研究
The Study of TGF-β Signaling in Normal Fibroblast Cell Line And Keloid Fibroblast Cell Line
摘要
中文摘要 蟹足腫是人類常見的疾病之一，是一種過度增生的疤痕組織，患者於創傷後，癒合的傷口超過原本皮膚的高度及傷口範圍，侵犯到周圍的正常組織。治療上相當困難，以傳統治療（手術及病灶內類固醇注射），復發率高達百分之五十。本病的制病機轉仍未明，細胞外成纖維細胞對細胞激素的反應及其下游的訊息傳遞基質膠原蛋白聚積可能為重要原因。 乙型轉型生長因子（TGF-β）可促進成纖維細胞的移行、增殖和膠原蛋白的製造，在許多慢性纖維化疾病當中，佔有相當重要的角色。 TGF-β主要相關的細胞內訊息傳導物質為Smads蛋白質，分為活化型轉錄因子Smad2、Smad3 和一般型轉錄因子Smad4及抑制型轉錄因子Smad6、Smad7。本實驗利用正常與蟹足腫成纖維細胞株施以各種濃度組合之TGF-β1投與方式，期望能找出其細胞內訊息傳遞及蛋白質的釋出之差異。 研究結果顯示正常成纖維細胞株之Smad2、Smad3、Smad4的表現沒有明顯的增加，而Smad6、Smad7有明顯增加。而蟹足腫成纖維細胞株的Smad2的表現有明顯的增加，Smad3、Smad4的表現沒有明顯的增加，Smad6、Smad7則沒有表現。綜合以上結果顯示乙型轉型生長因子對正常成纖維細胞株的作用是經由增加抑制型轉錄因子Smad6、Smad7來抑制活化型轉錄因子Smad2、Smad3，使之表現正常；而乙型轉型生長因子對蟹足腫成纖維細胞株而言，則是不經抑制型轉錄因子Smad6、Smad7的抑制調控，直接刺激活化型轉錄因子Smad2表現增加，故推測蟹足腫病變可能透過此一訊息傳遞之表現而增加細胞外基質蛋白的合成與堆積。
並列摘要
Abstract:Keloid is one of the most common skin disease in human beings.Which defined as scars growing beyond the confines of original woundsafter healing of a skin injury. There is no universally accepted effective treatment, with the conventional therapy, i.e. surgery and/or intralesional steroid injection，the recurrence rate is 50﹪.The pathogenesis is still unknow. The abnormalities in extracellular matrix collagen production, abnormal response of the fibroblasts to various cytokines and the signal transduction might be the important factors. TGF-β can induce cell migration、proliferation and production of collagen in fibroblasts, and play a key role in many chronic fibrotic disorders. Previous investigations revealed that Smads were involved in the TGF-β signaling in human fibroblasts. There are several homologs of Smad protein in the cell. Smad2 and Smad3 are receptor-regulated Smads .and Smad4 as the common mediator Smad and Smad6、7 as inhibitory Smads. The purpose of this study was designed by using the CCD966SK and KEL FIB cultures treated with different concentration of TGF-β1 to discover the regulatory mechanisms and the difference amount of releasing protein and gene expression. As the result, we discovered that in CCD966SK there was no apparent increased amount in Smad2, Smad3 and Smad4 expressed, but Smad6 、7 showed a significant increasing amount. For KEL FIB, the expression of Smad2 could be detected with increasing amount, but Smad3 and Smad4 had shown in small increasing amount. In contrast, Smad6 and Smad7 had no expression showed in this experiment. In conclusion, the effects of TGF-β on CCD966SK are in turn mediated by inhibition of Smad2 and Smad3 due to increase in Smad6 and Smad7. The effects of TGF-β on KEL FIB are in turn mediated activation of Smad2 without Smad6 and Smad7 regulation control. Therefore, the formation of Keloid might be caused by the regulatory mechanisms of TGF-β which stimulated extracellular matrix synthesis and deposition.