臺灣博碩士論文加值系統

　　本實驗室於前期研究中，證實中國南瓜的莖部含有降血糖的成分，但是中國南瓜其他組織的降血糖效果則不清楚。本研究的目的是比較中國南瓜莖、果實、種子三個不同組織的降血糖活性。實驗以Clone 9大鼠肝臟細胞與C2C12小鼠肌肉細胞為模型，分析南瓜莖、果實、種子的粗萃物是否促進正常細胞吸收葡萄糖。結果，三種粗萃物皆沒有明顯促進正常細胞吸收葡萄糖的效果，但是，進一步以粗萃物的分配萃取層進行分析，三種組織的乙酸乙酯萃取層皆可明顯促進正常細胞吸收葡萄糖，顯示三種組織皆含有類胰島素的活性天然物。進一步分析機制，發現三者皆可活化AMP-activated protein kinase (AMPK)及IRS-1的活化有關，顯示該萃取層的類胰島素效果來自兩種不同的分子機制。以TNF-誘導細胞產生胰島素抗性，以三種粗萃物處理，分析其降血糖效果，結果顯示，三種粗萃物具有胰島素增敏劑的效果。本實驗的結果證實，南瓜全株皆具有降血糖活性，包括胰島素類似物與胰島素增敏劑的功能，使用方式可先將莖、果實或種子先以乙醇萃取，再以乙酸乙酯進一步分配萃取，所得產物可用於第一型或第二型糖尿病的管理。

The stem of Cucurbita moschata has been confirmed to contain hypoglycaemic components, but the hypoglycaemic effects of other tissues of the plant are not clear. The purpose of this study is to investigate and compare the hypoglycaemic effects of the stems, the fruits, and the seeds of C. moschata. The rat normal hepatic cell line Clone 9 and the mouse muscle cell line C2C12 were used as models to assay whether the crude extracts of these tissues could promote the glucose consumption of the cells. Consequently, none of the crude extracts showed such an effect in these cells. However, after partitioning by organic solvents, the ethyl acetate (EA) layer of each of the three crude extracts promoted the glucose uptake of the cells, suggesting that the three tissues all contain molecules with insulin-like actities. Further analysis showed that the EA layers of the three tissues all activated AMP- activated protein kinase and increased the tyrosin phosphorylation of IRS-1, suggesting that the insulin-like of the EA layer is mediated by at least two different mechanisms. Subsequently, Clone 9 cells were treated with tumor necrosis factor-alpha (TNF-alpha) to induce insulin resistance, that were treated with the crude extracts of the three tissues. As a result, the crude extracts exhibited insulin sensitizing activites in insulin-resistant cells. Insummary, the stems, fruits, and seeds of C. moschata all contain hypoglycaemic activity, including insulin-like and insulin-sensitizing activities. It is suggested to extract the stems, fruits, or seeds with ethanol, followed by partitioning with acetate, the resulting product can be used manage type 1 and type 2 diabetes.

目錄
中文摘要 1
Abstract 2
誌謝 4
圖目錄 6
第1章 前言 1
1.1 研究背景 1
1.2 研究動機與架構 3
1.3 研究成果的重要性 4
第2章 文獻回顧 5
2.1 胰島素訊號傳遞路徑 5
2.2 胰島素抗性 (Insulin resistance) 7
2.3 AMPK 7
2.4治療糖尿病的藥物 9
2.5以細胞快速篩選降血糖分子的方法 10
第3章 材料與方法 11
3.1 實驗材料 11
3.1.1 南瓜粗萃物 11
3.1.2 哺乳動物細胞株 11
3.1.3 細胞培養基 12
3.1.4 試劑及緩衝溶液 12
3.1.5 蛋白質分子量標記 15
3.1.6抗體 (antibody) 15
3.2 實驗方法 16
3.2.1 細胞培養 16
3.2.2細胞葡萄糖吸收的分析 17
3.2.3 細胞存活率分析 (WST-1 assay) 17
3.2.4西方墨點法分析 (Western blot analysis) 17
3.3實驗數據統計分析 19
第4章 結果 20
4.1 南瓜粗萃物與萃取層的細胞毒性分析 20
4.2分析南瓜粗萃物是否提高細胞的葡萄糖吸收 20
4.3分析南瓜萃取層的活性 21
第5章 討論 33
第6章 結論 37
參考文獻 38
作者簡介 46

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