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研究生:陳嘉文
研究生(外文):Chia-Wen Chen
論文名稱:米麩油對第2型糖尿病大鼠體內胰島素敏感性與肝臟脂質代謝之影響
論文名稱(外文):Effects of Rice Bran Oil on Insulin Sensitivity and Hepatic Lipid Metabolism in Type 2 Diabetic Rats
指導教授:鄭心嫻鄭心嫻引用關係
學位類別:博士
校院名稱:臺北醫學大學
系所名稱:藥學系(博士班)
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:151
中文關鍵詞:米麩油第2型糖尿病大鼠高脂血症胰島素阻抗
外文關鍵詞:rice bran oiltype 2 diabeteshyperlipidemiainsulin resistanceSREBP-1c/ 2
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許多關於高脂血症的動物及人體研究報告,均指出攝取米麩油可顯著降低血漿膽固醇及三酸甘油酯濃度,但是對於給予米麩油的介入對體內血脂代謝相關酵素表現的調控機轉,以及對第2型糖尿病的影響,目前尚未完全釐清。本論文研究以nicotinamide / streptozotocin誘發第2型糖尿病大鼠模式進行實驗,在實驗一中以0 %、10 % 及15 % 米麩油含量的高油飼料餵養糖尿病大鼠四週,探討攝取米麩油對肝臟膽固醇代謝作用相關酵素mRNA表現的影響。在實驗二則以含15 % 米麩油之高油飼料餵養糖尿病大鼠四週,探討攝取米麩油對體內胰島素阻抗現象及肝臟膽固醇、脂質、葡萄糖代謝作用相關酵素和肌肉胰島素接受體蛋白質和mRNA表現的影響。並且更深入探討攝取米麩油是否會改變轉錄因子sterol regulatory element bindg protein-1c / 2 ( SREBP-1c、SREBP-2 )、carbohydrate response element binding protein ( ChREBP )、peroxisome proliferator activated receptor-??/?? ( PPAR-α、PPAR-?? ) 的表現,進而影響體內脂質代謝、糖質新生作用與葡萄糖利用等相關蛋白質的表現。
從實驗一的結果發現,第2型糖尿病大鼠以米麩油實驗飼料餵養四週後會降低禁食胰島素、血漿三酸甘油酯、低密度脂蛋白膽固醇與肝臟三酸甘油酯含量;並增加肝臟膽固醇合成酵素3-hydroxy-3-methylglutaryl coenzyme A reductase ( HMG-CoA reductase )、肝臟膽固醇代謝酵素cholesterol 7??-hydroxylase與肝臟low density lipoprotein-receptor ( LDL-receptor ) 的mRNA表現,增加肝臟膽固醇合成與代謝,造成糞便中性固醇及膽酸排出量增加,也首次發現米麩油可改善糖尿病大鼠的胰島素阻抗。在實驗二的研究結果中,第2型糖尿病大鼠以15 % 米麩油的實驗飼料餵養四週後,發現會增加轉錄因子SREBP-1c、SREBP-2的表現,並增加肝臟脂質合成酵素acetyl-CoA carboxylase、fatty acid synthase,以及HMG-CoA reductase、cholesterol 7??-hydroxylase、LDL-receptor和glucokinase的蛋白質與mRNA表現,造成糞便中性固醇及膽酸排出量增加。此外,從腹腔注射葡萄糖耐受試驗,與肌肉葡萄糖利用蛋白質insulin receptor與肌肉轉錄因子PPAR-?? 的表現結果,更證實攝取米麩油可以顯著改善糖尿病大鼠體內的胰島素阻抗現象。總結研究結果得知,攝取米麩油後會經由降低體內膽固醇儲存量,增加肝臟轉錄因子SREBP-1c、SREBP-2的表現,進而改變體內血脂代謝與葡萄糖利用蛋白質的表現,來增加糞便中性固醇與膽酸排出量,以及降低肌肉轉錄因子PPAR-?? 的表現,最後達到改善第2型糖尿病大鼠的胰島素敏感性及高血脂症狀。
The hypolipidemic and hypocholesterolic effects of rice bran oil (RO) consumption have been established in several hyperlipidemic animal and human studies. However, the regulatory mechanism by which RO affects lipid metabolic enzymes, as well as the effects of RO on insulin resistance and hyperglycemia in type 2 diabetes remains largely unknown. To investigate the effects of RO consumption on the expression of hepatic enzymes that influence cholesterol metabolism, streptozotocin/nicotinamide-induced type 2 diabetic rats were fed high-fat diets composed of 0, 10, and 15% RO for 4 weeks (Study 1). The effects of the RO consumption on insulin resistance in vivo and the protein expression of the hepatic enzymes that control lipid, glucose, and cholesterol metabolism were analyzed in type 2 diabetic rats that were fed a high-fat diet composed of 15% RO for 4 weeks ( Study 2 ). In addition, in study 2, the effects of RO consumption on sterol regulatory element binding protein-1c/2 ( SREBP-1c/2 ), carbohydrate response element binding protein ( ChREBP ), and peroxisome proliferator activated receptor-??/?? ( PPAR-α/?? ), which are transcription factors that influence the expression of the hepatic enzymes and muscular proteins regulating glucose uptake, was analyzed. In study 1, RO decreased fasting plasma insulin, triglyceride and LDL cholesterol levels in diabetic rats fed high-fat diet. In addition, RO consumption decreased hepatic triglyceride levels and increased 3-hydroxyl-3- methyl-glutaryl-CoA ( HMG-CoA ) reductase, cholesterol 7??- hydroxylase, and LDL - receptor mRNA expression. Finally, RO consumption increased excretion of fecal bile acids and neural sterols. In study 2, RO consumption increased expression of SREBP-1c and SREBP-2 transcription factors, which further increased hepatic acetyl-CoA carboxylase, fatty acid synthase, HMG-CoA reductase, cholesterol 7??-hydroxylase, LDL-receptor, and glucokinase protein and mRNA expression in diabetic rats. After RO consumption, increased excretion of fecal bile acids and neural sterols were detected in diabetic rats. In addtion, intraperitoneal glucose tolerance tests and analysis of the muscular insulin receptor and PPAR-?? protein expression revealed that ingestion of RO improved the insulin sensitivity in diabetic rats. In conclusions, ingestion of RO depletes the cholesterol pool in vivo and further affects SREBP-1c and SREBP-2 expression, resulting in altered mRNA and protein expression of lipid and cholesterol metabolic enzymes in liver, and increases excretion of fecal bile acids and neural sterols. Furthermore, RO consumption decreases the expression of muscular PPAR-??. Finally, RO consumption improves the insulin resistance and hyperlipidemia in type 2 diabetic rats.
目錄
誌謝 I
中文摘要 III
Abstract V
目錄 VII
表目錄 IX
圖目錄 X
Abbreviations XI
第一章 前言 1
第二章 文獻回顧 3
第一節 糖尿病 3
一、糖尿病盛行率與死亡率現況 3
二、糖尿病的定義與分類 3
三、胰島素訊息傳遞與胰島素阻抗 4
四、葡萄糖轉運蛋白 6
五、糖尿病高脂血症的機轉 7
六、糖尿病的治療 8
第二節 肝臟醣類及脂質代謝的影響 11
一、肝臟與血糖的恆定 11
二、肝臟葡萄糖儲存作用 11
三、肝臟葡萄糖生成作用 12
四、肝臟的脂肪酸合成與膽固醇代謝 13
五、胰島素對肝臟合成、代謝醣類及脂質相關酵素的影響 14
六、調控肝臟脂肪酸合成與膽固醇代謝的轉錄因子 15
第三節 脂肪酸 18
一、脂肪酸的分類與命名 18
二、飲食脂質總量與種類對血脂質的影響 19
三、飲食脂質種類對血脂質的影響 20
四、飲食中脂肪酸對血脂質及胰島素抗性的影響 20
第四節 米麩油 21
一、米麩油的組成分 21
二、植物固醇 21
三、Gamma-oryzanol 22
四、Alpha-tocopherol及gamma-tocotrienol 23
五、米麩油對血脂質的影響 24
第三章 實驗一攝取米麩油對第2型糖尿病大鼠肝臟膽固醇與脂質代謝酵素的影響 26
第一節 實驗目的 26
第二節 實驗材料與方法 26
第三節 實驗結果 37
一、體重與攝食量變化 37
二、禁食血漿葡萄糖及脂質濃度變化 37
三、肝臟重量及脂質含量變化 38
四、血漿脂肪酸組成的變化 38
五、肝臟脂肪酸組成的變化 39
六、糞便膽酸和中性固醇排出量的變化 40
七、肝臟中膽固醇代謝相關酵素的mRNA表現量 40
第四節 討論 41
第五節 結論 46
第四章 實驗二米麩油對第2型糖尿病大鼠體內胰島素敏感性、脂質與葡萄糖代謝酵素蛋白質表現的影響 47
第一節 實驗目的 47
第二節 實驗材料與方法 47
第三節 實驗結果 55
一、體重及攝食量變化 55
二、禁食血漿葡萄糖及脂質濃度變化 55
三、腹腔注射葡萄糖耐受試驗血漿葡萄糖與胰島素濃度變化 55
四、肝臟重量及脂質含量變化 56
五、血漿脂肪酸與肝臟脂肪酸組成的變化 56
六、糞便膽酸和中性固醇排出量的變化 57
七、肝臟的脂質與膽固醇合成代謝相關酵素mRNA與蛋白質表現量 57
八、肝臟的糖解與糖質新生作用相關酵素mRNA與蛋白質表現量 58
九、肝臟轉錄因子ChREBP、SREBP-1c、及PPAR-?? mRNA和蛋白質表現量 59
十、肌肉胰島素接受體mRNA和蛋白質表現量 59
十一、肌肉轉錄因子PPAR-?? mRNA和蛋白質表現量 59
第四節 討論 60
第五章 總結論 69
參考文獻 70
附錄一 Tocopherols and tocotrienols 分子結構式 143
附錄二 ??-oryzanol成份的分子結構式 144
附錄三 實驗試劑及設備 145
附錄四 已發表著作 147
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