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研究生: 曾雅旻
Ya-Ming Tseng
論文名稱: 山苦瓜對apoE剔除鼠脂質代謝之影響
The effects of wild bitter gourd on lipid metabolism
指導教授: 吳文惠
Wu, Wen-Huey
學位類別: 碩士
Master
系所名稱: 人類發展與家庭學系
Department of Human Development and Family Studies
論文出版年: 2006
畢業學年度: 94
語文別: 中文
論文頁數: 105
中文關鍵詞: 山苦瓜apo E剔除小鼠過氧化體增殖劑活化受器α微粒體三酸甘油脂轉運蛋白
英文關鍵詞: wild bitter gourd, apolipoprotein E knockout mice, PPARα (perioxisome proliferators activated receptor- alpha), MTP (microsomal triglyceride transfer protein)
論文種類: 學術論文
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  • 代謝症候群是指胰島素抗性所引起的葡萄糖不耐症、腹部肥胖、高TG、低HDL與高血壓等症狀,而最終將有可能會轉變為心血管疾病,對健康造成更大的傷害。在印度、中國及拉丁美洲地區,苦瓜被用做改善糖尿病的藥物已有很長久的歷史,但苦瓜對脂質代謝影響的研究則是近幾年才開始發展。本研究以血脂異常且容易產生動脈粥狀硬化之缺乏apoE小鼠為模式,探討苦瓜改善脂質代謝情形。實驗一、18週大缺乏apoE小鼠分為6組,給予不同飼料如下:C組(n = 5)為一般chow diet、HF組(n = 10)含20% fat、BGP組(n = 10)含20% fat加5%山苦瓜凍乾粉末、EAE組(n = 10)含20% fat加1%山苦瓜乙酸乙酯萃物、CF組(n = 5)含20%fat加0.5%clofibrate、TZD組(n = 5)含20%fat加0.00123% rosiglitazone,共餵養18週後犧牲,其間於第4、9、17週時採眼窩血測量血脂變化,並於犧牲後取組織器官進行後續分析。實驗結果以單因子變異數分析及Tukey事後分析進行組間比較。結果BGP組第9及17週血液TC顯著較HF組低25及39% (p<0.05)、第17週血液TG較HF組低38% ( p<0.05)、HDL-C較HF組高55%( p<0.05),EAE組則使血液HDL-C高於HF組39%但未達顯著差異,肝臟脂質各組間無顯著差異。實驗二、改用較年輕缺乏apoE小鼠,並重新萃取山苦瓜乙酸乙酯萃物,分為3組:HF組(n = 11)、BGP組(n = 9)、EAE組(n = 10),飼料成分同前述,餵食8週後,BGP組TC較HF組低28%( p<0.05)、TG下降幅度顯著高於HF組( p<0.05)、血液HDL-C高於HF組25%,但未達顯著差異。血液adiponectin、leptin濃度、肝臟PPAR-α、MTP mRNA表現量,各組間無顯著差異。本研究結果顯示,山苦瓜凍乾粉末對apo E剔除小鼠具有改善血脂功效,可研發為降血脂保健食品。

    The apolipoprotein E knockout (apoE-/-) mice are hyperlipidemic and a good model to study atherosclerosis. In order to know the effects of wild bitter gourd on lipid metabolism, we conducted two experiments in apoE-/- mice. In experiment 1, 18-wk-old apoE-/- mice were assigned to 6 groups and fed different diets as follows: the high fat (20%, w/w) diet (HF, n = 10), HF with 5%wild bitter ground powder (BGP, n = 10), HF with 1% ethyl acetate extract of bitter gourd (EAE, n = 10), HF with 0.5% clofibrate (CF, n = 5), HF with 0.00123% rosiglitazone (TZD, n = 5) and chow diet (C, n = 5). Serum total cholesterol (TC) and triglyceride (TG) concentrations were measured at the end of wk 4, 9 and 17. After 18 wk of feeding, the mice were sacrified. Liver and blood samples were obtained to determine hepatic TC, TG and serum HDL-C concentrations. Data were analyzed by one-way analysis of variance and the Tukey post hoc. The BGP group had a lower level of serum total cholesterol in week 9 and 17 (25 & 39% reduction, respectively, p<0.05), a lower level of serum triglyceride in week 17 (38% reduction, p<0.05) and a higher level of HDL-C in wk 18 (55% increase, p<0.05) than the HF group. The EAE group had a 39% higher level of HDL-C than the HF group in week 18, but it was not statistically significant. Levels of hepatic TC and TG were not significantly different among all groups. In experiment 2, young (6-10 wk-old) apoE-/- mice were assigned to 3 groups and fed the HF diet (n = 11), the BGP diet (n = 9) and the EAE diet (n = 10), respectively, for 8 wk. After feeding trials, the BGP group had a lower level of serum total cholesterol (28% reduction, p<0.05), and a larger decrease in level of serum triglyceride in wk 8 (p<0.05) than the HF group. The BGP group also had a 25% higher level of HDL-C than HF group, but it was not statistically significant. There were no significant difference in serum adiponectin, serum leptin, liver PPAR-αmRNA expression and liver MTP mRNA expression among HF、BGP and EAE groups. In conclusion, this study shows that wild bitter gourd powder can be a potent blood lipid-lowering health food.

    縮寫對照表 組別縮寫意義 中文摘要 英文摘要 目錄 第一章 緒論 1 第二章 文獻探討 3 第一節 苦瓜介紹 3 一.名稱 3 二.植物性狀 3 三.生產分布與栽培環境 4 四.食用特性 5 五.山苦瓜介紹 5 六.苦瓜生理效用 6 苦瓜對血脂、肝脂的影響 7 苦瓜對脂質代謝影響的可能機制 13 a.關於PPAR 13 簡介PPAR 13 苦瓜與PPAR 15 b.關於MTP (Microsomal triglyceride transfer protein) 17 VLDL合成模式假說 17 苦瓜與MTP 19 第二節ApoE knockout mice介紹 19 一.小鼠與人類血脂異同 19 二.Apolipoprotein E 21 三.ApoE knockout mice 21 四.ApoE knockout mice脂質代謝變化 22 第三節 脂肪細胞激素──adiponectin與leptin 24 Adiponectin 24 Leptin 25 第四節 研究動機與目的 26 第三章 實驗一 27 第一節 材料與方法 27 一.實驗流程大綱 27 二.山苦瓜樣品製備 27 三.飼料配方製備 27 四.動物飼養 31 五.血液樣本收集 31 六.動物犧牲 32 七.血液脂質分析 32 八.肝臟脂質分析 33 九.統計分析 34 第二節 結果 35 一.生長情形 35 二.組織器官絕對重量與相對重量 38 三.血液脂質濃度 40 (一) 總膽固醇濃度 40 (二) 三酸甘油酯濃度 42 (三) 高密度脂蛋白膽固醇濃度 44 四.肝臟脂質濃度 45 (一) 總膽固醇濃度 45 (二) 三酸甘油酯濃度 45 第三節 討論 46 第四章 實驗二 51 第一節 材料與方法 52 一.實驗流程大綱 52 二.山苦瓜樣品製備 52 三.飼料配方製備 52 四.動物飼養 53 五.血液樣本收集 53 六.動物犧牲 54 七.血液脂質分析 54 八.血液leptin分析 54 九.血液adiponectin分析 55 十.半定量反轉錄聚合酶鏈鎖反應 55 十一.統計分析 58 第二節 結果 59 一.生長情形 59 二.組織器官絕對重量與相對重量 62 三.血液脂質濃度 63 (一) 總膽固醇濃度 63 (二) 三酸甘油酯濃度 63 (三) 高密度脂蛋白膽固醇濃度 64 四.血液adiponectin濃度 65 五.血液leptin濃度 66 六.肝臟mRNA表現 67 (一)PPAR-α 67 (二)MTP 68 七.相關性分析 69 (一) 血液總膽固醇濃度 69 (二) 血液三酸甘油酯濃度 71 (三) 血液adiponectin濃度 73 (四) 血液leptin濃度 76 第三節 討論 78 第五章 綜合討論 82 第六章 結論 86 參考文獻 87 表目錄 表2-1 近五年苦瓜果實對實驗動物脂質代謝影響文獻彙整.............................. 8 表2-2 小鼠與人類脂蛋白生理不同點 20 表2-3 ApoE剔除小鼠與一般小鼠及人類血液膽固醇比較 22 表3-1-1適應期飼料配方 28 表3-1-2 山苦瓜成份分析 29 表3-1-3 實驗期飼料配方 30 表3-2-1 ApoE剔除小鼠餵食實驗飼料17週體重增加量、平均攝食量及攝食效應 37 表3-2-2 ApoE剔除小鼠餵食實驗飼料17週後組織絕對及相對重量 39 表3-2-3 ApoE剔除小鼠餵食實驗飼料17週實驗期間血液總膽固醇 41 表3-2-4 ApoE剔除小鼠餵食實驗飼料17週實驗期間血液三酸甘油酯 43 表3-2-5 ApoE剔除小鼠餵食實驗飼料17週後肝臟中膽固醇、三酸甘油酯濃度 45 表4-2-1 ApoE剔除小鼠餵食實驗飼料8週體重增加量、平均攝食量及攝食效應 61 表4-2-2 ApoE剔除小鼠餵食實驗飼料8週後組織絕對及相對重量 62 表4-2-3 ApoE剔除小鼠餵食實驗飼料8週實驗期間血液總膽固醇、三酸甘油酯濃度 63 表4-2-4 實驗第8週血液總膽固醇濃度與各因子相關性及排除體重、體重變化、副睪脂、腎周脂或副睪脂及腎周脂後淨相關 70 表4-2-5 實驗第8週血液三酸甘油酯濃度與各因子相關性及排除體重、體重變化、副睪脂、腎周脂或副睪脂及腎周脂後淨相關 72 表4-2-6 血液adiponectin濃度與各因子相關性及排除體重、體重變化、副睪脂、腎周脂或副睪脂及腎周脂後淨相關 74 表4-2-7 血液leptin濃度與各因子相關性及排除體重、體重變化、副睪脂、腎周脂或副睪脂及腎周脂後淨相關 77 表5-1 本研究實驗一、二結果整理比較 85 圖目錄 圖2-1 市售苦瓜 4 圖2-2 改良種山苦瓜花蓮一號 6 圖2-3 PPARs作用機制 14 圖2-4 VLDL合成模式假說 18 圖3-2-1 ApoE knockout mice 餵食實驗飼料17週生長曲線 36 圖3-2-2 ApoE knockout mice 餵食實驗飼料17週後血中高密度脂蛋白膽固醇濃度 44 圖4-2-1 ApoE剔除小鼠餵食實驗飼料8週生長曲線 60 圖4-2-2血液HDL-C濃度 64 圖4-2-3血液adiponectin濃度 65 圖4-2-4血液leptin濃度 66 圖4-2-5餵食實驗飼料8週後以半定量反轉錄聚合酶鏈鎖反應偵測apoE剔除小鼠肝臟組織PPAR-α mRNA表現量 67 圖4-2-6餵食實驗飼料8週後以半定量反轉錄聚合酶鏈鎖反應偵測apoE剔除小鼠肝臟組織MTP mRNA表現量 68

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