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系統識別號 U0026-2210201310015400
論文名稱(中文) 香椿萃取物對脂肪細胞脂質代謝調控機制之探討
論文名稱(英文) Study of Toona sinensis extracts on lipid metabolism in adipocytes and adipose tissue
校院名稱 成功大學
系所名稱(中) 生命科學系
系所名稱(英) Department of Life Sciences
學年度 102
學期 2
出版年 103
研究生(中文) 蔡岳騰
研究生(英文) Yueh-Teng Tsai
學號 L56991021
學位類別 碩士
語文別 中文
論文頁數 102頁
口試委員 指導教授-張素瓊
口試委員-王浩文
口試委員-尤文正
中文關鍵字 香椿  過氧化小體增生活化受體  脂肪細胞  脂肪酸氧化 
英文關鍵字 Toona sinensis Roem  peroxisome proliferator activated receptorα  adipocyte  fatty acid oxidation 
學科別分類
中文摘要 民間傳統使用香椿(Toona sinensis Roem)來調降血糖與血脂。研究指出餵食香椿葉萃取物給予高血脂症小鼠,顯著降低小鼠血清總膽固醇、三酸甘油脂(TG)、低密度脂蛋白(LDL)、增加高密度脂蛋白(HDL),且降低LDL/HDL的比值,然而其降血脂分子機制仍未明瞭。因此本研究主要探討香椿萃取物 (Toona sinensis leaf extract, TSLE)對高脂飲食 (High Fat Diet, HFD)小鼠胰島素敏感性、脂肪細胞分化及脂質代謝相關蛋白表現量的影響。使用TSLE分別處理分化初期與分化後成熟的3T3-L1脂肪細胞,,以 Western blotting檢測分化及脂質代謝相關蛋白的表現、Real-Time PCR檢測分化相關基因、脂質生成相關基因與脂肪酸分解、氧化代謝相關基因的表現、偵測葡萄糖消耗率、觀察脂質累積、檢測活化PPARα活性,探討TSLE對脂肪細胞脂質代謝的調控。同時以HPLC進行TSLE成分鑑定,利用luciferase assay篩選成分活化PPARα活性,將有活化PPARα功能的成分處理3T3-L1,探討各成分對脂質代謝的調控。結果顯示,TSLE經由調節脂肪細胞脂質分解及脂肪酸氧化代謝路徑降低脂質累積,可開發成降血脂或減少體脂肪累積功能之保健產品。
英文摘要 In the present study, HFD mice and 3T3-L1 adipocytes were used to investigate the mechanism of Toona sinensis leaf extrcts (TSLE) on lipid metabolism. High-perfomance liquid chromatography (HPLC) and PPARα-LBD luciferase assay were used to investigate the functional compounds of TSLE. Results showed that TSLE decreased lipid accumulation in adipocyte via the regulation of lipolysis and fatty acid oxidation, suggesting that TSLE is potential to develop into a function food for hypolipidemia and decreased body-fat accumulation.
論文目次 考試合格證明 I
摘要 II
Abstract III
致謝 V
目錄 VI
表目錄 XI
圖目錄 XII
前言 1
文獻探討 4
肥胖的定義 4
肥胖與胰島素阻抗 5
代謝症候群 6
代謝症候群與疾病 6
代謝症候群與血脂異常 7
脂肪細胞 8
脂質的運輸 8
脂質的合成與分解 10
脂肪酸的合成 11
脂肪酸分解 12
脂肪酸氧化 β-oxidation 12
第四型葡萄糖轉運子 GLUT4 13
PPAR pathway 14
PPAR之種類 15
PPAR之結構 15
PPAR之活化劑與ligands 16
PPAR之標的基因 18
PPARα在脂肪細胞的研究 19
Adiponectin 20
香椿 21
國內外香椿研究 22
香椿葉酒精萃取物(TSLE)先前研究 23
材料與方法 24
細胞株 24
藥品 24
抗體 28
primer 28
常用buffer 29
實驗儀器 31
實驗架構 35
HFD動物模式 35
細胞模式 36
實驗動物 37
實驗方法 38
口服葡萄糖耐受性試驗 (oral glucose tolerance test, OGTT) 38
3T3-L1脂肪細胞的分化 38
MTT assay 38
油紅染色 39
蛋白質萃取 39
蛋白質定量 39
膜蛋白萃取 40
Total RNA萃取 40
DNA decontamination 41
反轉錄作用(Reverse Transcription) 41
Plasmid 的萃取 41
蛋白質電泳 42
蛋白質轉漬 42
免疫呈色 43
Real-Time PCR (Q-PCR) 43
Transfection 43
PPARα-LBD luciferase assay 44
Liquid-liquid partition分離TSLE 44
HPLC分析TSLE 45
TSLE成分定量 45
脂肪酸衍生化 45
脂肪酸衍生物分析 46
TSLE中脂肪酸成分定量 46
統計方式 46
結果 47
TSLE對HFD小鼠口服葡萄糖耐受性的影響 47
TSLE對HFD小鼠脂肪組織PPARα及adiponectin 蛋白表現的影響 47
TSLE活化PPARα評估 47
TSLE對分化初期3T3-L1細胞脂質累積的影響 48
TSLE對分化初期3T3-L1細胞分化與成熟蛋白表現的影響 48
TSLE對分化初期3T3-L1細胞分化與脂質代謝相關基因表現的影響 48
PPARα拮抗劑對分化初期3T3-L1細胞脂肪累積的影響 48
PPARα拮抗劑對分化初期3T3-L1細胞分化與脂質代謝相關基因表現的影響 49
TSLE對分化初期3T3-L1細胞葡萄糖利用率與細胞膜上GLUT4蛋白表現的影響 49
TSLE對分化後成熟3T3-L1細胞脂質累積的影響 50
TSLE對分化後成熟3T3-L1細胞分化與成熟相關蛋白表現的影響 50
TSLE對分化後成熟3T3-L1細胞脂質代謝相關基因表現的影響 50
PPARα拮抗劑對分化後成熟3T3-L1細胞脂肪累積的影響 50
PPARα拮抗劑對分化後成熟3T3-L1細胞基因表現的影響 51
TSLE液相分配萃取物活化PPARα效果評估 51
TSLE已知成分活化PPARα活性評估 51
Gallic acid與Rutin對分化初期T3-L1細胞脂肪累積的影響 52
Gallic acid與Rutin對分化初期T3-L1細胞分化與成熟相關蛋白表現的影響 52
Gallic acid與Rutin對分化初期T3-L1細胞脂質代謝相關基因表現的影響 52
TSLE中脂肪酸的組成 53
TSLE與香椿75 %酒精萃取物 (TSL-E5)成分差異 53
香椿葉各種酒精濃度萃取物活化PPARα活性 54
TSLE中脂肪酸成分活化PPARα活性 54
TSLE中脂肪酸成分對分化後成熟3T3-L1脂肪細胞基因表現的影響 55
TSLE中脂肪酸成分對分化後成熟3T3-L1脂肪細胞脂質累積的影響 55
討論 56
結論 62
參考文獻 63
圖 75
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