- 學位論文
黃花蜜菜對TPA誘導上皮細胞轉型之影響研究
Study of Wedelia chinensis on TPA-induced epidermal cell transformation
摘要
天然植物中富含的多酚成分具有抗氧化，抗癌及抗發炎的功效。此外，慢性發炎被發現與癌症形成具有密切的相關性。因此，我們萃取黃花蜜菜 (Wedelia chinensis)中具抗氧化的成分，了解是否具有抵抗化學物質誘導的細胞轉型能力。首先，將黃花蜜菜以酒精萃取出粗萃物後，以正己烷、乙酸乙酯(EA)及純水，將其分配萃取。比較其中活性成分及抗氧化能力後，發現EA層的能力最佳。在細胞實驗部分，使用JB6 P+細胞 (老鼠promotion-sensitive上皮細胞)以12-O-tetradecanoylphorbol-13-acetate (TPA)誘導轉型進行非固著依賴性生長，在經過42天培養後，發現有預先處理EA層的細胞較單獨處理TPA細胞轉型惡化的情形較低。以動物實驗做更進一步的觀察，使用TPA塗抹於老鼠背部，使其產生大量的Reactive oxygene species (ROS)、表皮增厚、角質化。而有預先塗抹EA層的老鼠可有效降低其背部表皮的增生、發炎及角質化的情形，綜合上述結果，我們推測黃花蜜菜EA層具有抵抗化學物質所誘發轉型作用的能力。
並列摘要
The nature plants are rich in the polyphenolic phytochemicals that have the functions of antioxidant, anti-tumor, and, anti-inflammation. In addition, the chronic inflammation has a close relationship to carcinogenesis. To clarify the anti-oxidative components of Wedelia chinensis that has the potential of anti-transformation by chemicals induced, first, Wedelia chinensis was extracted with Ethanol. The crude extract was partitioned by n-Hexane, Ethyl acetate (EA), and pure water. We found the EA fraction has the highest anti-oxidation capacity by evaluation with total phenolic content, 1,1-diphenyl-1-2-picryl-hydrazyl (DPPH) assay, and xanthine oxidase activity assay. For the following study, by anchorage-independent transformation assay, we used the TPA induced JB6 P+ cells transformation. After 42 days, we found these cells that pre-treated by EA fraction possessed lower transformation ability than that TPA treated alone. In animal study, we used TPA on mouse skin to induce skin thickness and hyperkeratosis. As a result, the mouse that pre-treated by EA fraction decreased the epidermal skin layer number, inflammation, hyperkeratosis. In conclusion, our data presented the EA fraction of Wedelia chinensis has a potential of anti-transformation.