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研究生:黃郁婷
研究生(外文):Yuh-Ting Huang
論文名稱:巴西洋菇子實體及其液態發酵產物對大鼠血糖之影響
論文名稱(外文):Effects of Agaricus blazei and its liquid fermentation product on plasma glucose in rats
指導教授:孫璐西孫璐西引用關係
指導教授(外文):Lucy-Sun Hwang
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:食品科技研究所
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:103
中文關鍵詞:糖尿病代謝症候群巴西洋菇口服葡萄糖耐受性試驗
外文關鍵詞:Diabetes mellitusmetabolic syndromeAgaricus blazeioral glucose tolerance test
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糖尿病已成為一個重要的全球性健康議題。近年研究發現,包含高血壓、高胰島素血症、胰島素阻抗、血脂代謝異常和肥胖等諸多病症的代謝症候群 (metabolic syndrome),與糖尿病及心血管疾病關係密切,已成為各國研究及預防的重點。巴西洋菇(Agaricus blazei)具有抗腫瘤、抗氧化、增強免疫力及護肝等功效,但是對抗糖尿病之探討較少;此外,人工栽培巴西洋菇成本高、產量低且品質不穩,故以液態深層發酵 (submerged fermentation) 生產巴西洋菇被認為是值得開發的生產方式,其保健功效如何則有待進一步評估。
本研究之目的在探討巴西洋菇子實體及以含豆科植物為發酵基質的巴西洋菇液態發酵產物對於血糖之影響。實驗分為急性試驗及長效性試驗兩部分。在急性試驗的第一階段,將巴西洋菇子實體及其液態發酵產物之熱水萃出物,分別對正常鼠、代謝症候群鼠及糖尿病鼠進行口服葡萄糖耐受性試驗,藉此評估兩種樣品熱水萃出物對血糖之影響,並篩選出有效樣品及其劑量。第二階段則將有效樣品與當作正控制組的關華豆膠做降血糖效果之比較。長效性試驗方面,則以高果糖飼料誘發大鼠產生代謝症候群症狀的模式,探討餵食巴西洋菇子實體及其液態發酵產物是否具有改善代謝症候群諸多症狀的效果。
急性試驗結果顯示,在正常鼠之口服葡萄糖耐受性試驗中,給予0.56 g/kg BW 的液態發酵產物熱水萃出物之大鼠,在灌糖後第 30 分鐘的血糖值顯著較對照組為低 (p<0.05),在第 120 分鐘時則明顯高於對照組 (p<0.01),顯示液態發酵產物熱水萃出物能有效減緩血糖上升;但在代謝症候群鼠則以 0.28 g/kg BW 的劑量在第 90 分鐘顯著降低血糖值 (p<0.05);糖尿病鼠方面,也是給予 0.28 g/kg BW 的液態發酵產物熱水萃出物能在第 30 分鐘展現降血糖功效 (p<0.05)。另一方面,巴西洋菇子實體熱水萃出物在三種動物模式之口服葡萄糖耐受性試驗中,不論是給予 0.28 g/kg BW 或 0.56 g/kg BW 的劑量皆無法影響血糖變化。因此選用液態發酵產物熱水萃出物進入第二階段實驗。由第二階段結果發現,液態發酵產物熱水萃出物的確具有減少血糖上升的效果,且效果與關華豆膠 (0.0375 g/kg BW) 相比,兩者效果相當,皆能延緩血糖的上升。
長效性試驗結果顯示,飼料中添加 10 % 巴西洋菇子實體時,可改善由高果糖飲食所誘發之高血壓、高三酸甘油酯血症、高血糖及高胰島素血症等現象,在口服葡萄糖耐受性試驗中亦可顯著降低血糖及提升胰島素敏感性,且明顯降低肝中三酸甘油酯、總膽固醇及 TBARS 之濃度,顯示巴西洋菇子實體具有減輕代謝症候群諸多症狀的效果。飼料中添加 5 % 液態發酵產物時,亦具有改善高三酸甘油酯血症及高血壓之效果,並能減少肝中三酸甘油酯與總膽固醇的堆積,但在口服葡萄糖耐受性試驗中,降低血糖及提升胰島素敏感性的效果則不如巴西洋菇子實體。
Diabetes mellitus (DM) has become a major global health problem. Recent research finding have demonstrated that there is a strong association among metabolic syndrome, DM and cardiovascular disease (CVD). Metabolic syndrome is a cluster of metabolic abnormalities including hypertension, hyperinsulinemia, insulin resistance, dyslipidemia and obesity. Therefore, prevention of metabolic syndrome is an important public health issune in many countries. Agaricus blazei has been shown to possess many beneficial health effects, including antitumor activity, antioxidation, immune enhancement and liver protection. But little is known about its antidiabetic activity. The prime cost of cultivating A.blazei is high and the yield is low with unstable quality. Therefore, using submerged fermentation technology to produce A. Blazei has been considered to be a method worth developing. Its health protection effect, however, needs to be confirmed.
The purpose of this study was to investigate the effects of A. Blazei and its liquid fermentation product on plasma glucose in rats. The study contains two parts, the acute and the long term experiments. At the first stage of the acute experiment, we used hot water extracts of A. Blazei (AB-hw) and its liquid fermentation product (ABFP-hw) in the oral glucose tolerance test (OGTT) to normal rats, metabolic syndrome rats and diabetic rats in order to evaluate the hypoglycemic effects of these two kinds of hot water extracts and find out the more effective one and its dose. At the second stage, the hypoglycemic effect of the chosen one was compared with guar gum which was designed as the positive control group. In the long term experiment, we investigated the effect of the fruiting body of A. Blazei and its liquid fermentation product on the symptoms of metabolic syndrome in rats which were induced by high fructose diet.
Results of the acute experiment showed that plasma glucose levels of the normal Sprague-Dawley rats administered 0.56 g/kg BW of ABFP-hw were lower than the control group after 30 minutes of glucose ingestion (p<0.05) but were higher than the control group after 120 minutes (p<0.01). This indicated that ABFP-hw could effectively delay the rise of plasma glucose. In metabolic syndrome rats and diabetic rats, 0.28 g/kg BW ABFP-hw administration significantly reduced plasma glucose after 90 minutes (p<0.05) and 30 minutes (p<0.05) of glucose ingestion. On the other hand, AB-hw had no effect on plasma glucose changes of OGTT for all these animal models. Therefore, we chose ABFP-hw for further study. In the second stage, ABFP-hw was found again to have hypoglycemic effect which was similar to the effect of gaur gum. Both of them could delay the elevation of plasma glucose on OGTT.
In the long term experiment, we found that rats fed with 10% A.Blazei could ameliorate the hypertension, hypertriglyceridemia, hyperglycemia and hyperinsulinemia induced by high fructose diet. A.Blazei added at 10% level in the deit could also reduce plasma glucose, increase insulin sensitivity in the OGTT test and decrease the concentrations of triglyceride, total cholesterol and TBARS in rat liver. These results indicated that A.Blazei could ameliorate the metabolic syndrome. Liquid fermentation product of A.blazei (ABFP) added at 5% level in the deit also had effects on ameliorating hypertriglyceridemia and hypertention and could also decrease the accumulation of liver triglyceride and total cholesterol. Comparing with the fruiting bodies of A.blazei, the liquid fermentation product had less efficacy in reducing plasma glucose and increasing insulin sensitivity on OGTT.
中文摘要………………………………………………………………………………...i
英文摘要………………………………………………………………………...……...iii
目錄……………………………………………………………………………………...v
圖次……………………………………………………………………………………...x
表次……………………………………………………………………...……………..xii

第一章 前言………………………………………………………………………....1
第二章 文獻回顧……………………………………………………………………2
第一節 糖尿病………………………………………………………………….. 2
一、 糖尿病的定義與症狀……………………………………………………..2
二、 糖尿病的診斷及分類……………………………………………………..3
三、 糖尿病的併發症…………………………………………………………..6
第二節 胰島素…………………………………………………………………...7
一、 簡介……………………………………………………………………...7
二、 胰島素接受器…………………………………………………………...7
三、 胰島素對糖類代謝的影響……………………………………………...8
四、 胰島素對脂肪代謝的影響……………………………………………...8
五、 胰島素對蛋白質代謝的影響…………………………………………...9
六、 胰島素阻抗……………………………………………………………...9
第三節 代謝症候群…………………………………………………………….10
一、 代謝症候群的定義與診斷…………………………………………….10
二、 代謝症候群形成之可能機制………………………………………….12
三、 代謝症候群與常見慢性病之關係.........................................................13
第四節 動物模式……………………………………………………………….14
一、 以高果糖飲食誘發代謝症候群……………………………………….14
二、 以streptozotocin誘發糖尿病…………………………………………..14
第五節 巴西洋菇……………………………………………………………….16
一、 巴西洋菇之發現與由來……………………………………………….16
二、 巴西洋菇之分類及型態特徵………………………………………….16
三、 巴西洋菇之組成……………………………………………………….16
四、 巴西洋菇之安全性…………………………………………………….18
五、 巴西洋菇之生理活性………………………………………………….18
第六節 關華豆膠……………………………………………………………….20
一、 來源與應用…………………………………………………………….20
二、 分子結構與溶解度…………………………………………………….20
三、 關華豆膠之生理活性………………………………………………….21
第三章 研究目的與實驗架構……………………………………………………….23
第一節 研究目的……………………………………………………………….23
第二節 實驗架構……………………………………………………………….24
實驗I、急性試驗
探討急性灌食巴西洋菇子實體及其液態發酵產物之熱水萃出物對大鼠葡萄糖耐受性的影響…………………………………………..24
實驗II、長效性試驗
探討連續餵食巴西洋菇子實體及其液態發酵產物對高果糖飲食誘發代謝症候群之大鼠的影響……………………………………….25
第四章 材料與方法………………………………………………………………….30
第一節 實驗材料……………………………………………………………….30
一、 巴西洋菇……………………………………………………………….30
二、 巴西洋菇液態發酵產物……………………………………………….30
三、 巴西洋菇子實體及其液態發酵產物之熱水萃出物………………….30
四、 關華豆膠……………………………………………………………….31
第二節 化學藥品、酵素套組與動物飼料……………………………………..31
第三節 儀器設備……………………………………………………………….34
一、 用於材料之一般成分分析…………………………………………….34
二、 用於實驗材料及動物飼料之製備…………………………………….34
三、 用於動物實驗樣品之收集與分析…………………………………….35
第四節 實驗方法……………………………………………………………….36
一、 一般成份分析………………………………………………………….36
二、 萃出率之測定………………………………………………………….40
三、 還原醣之測定………………………………………………………….40
四、 動物實驗……………………………………………………………….41
實驗I、 急性試驗 ………………………………………………………41
(一) 急性試驗 (A) 正常鼠…………………………………………42
(二) 急性試驗 (B) 代謝症候群鼠…………………………………43
(三) 急性試驗 (C) 糖尿病鼠………………………………………44
實驗II、長效性試驗……………………………………………………..45
五、 動物實驗之測定……………………………………………………….46
(一) 血壓之測量.................................................................................47
(二) 口服葡萄糖耐受性試驗.............................................................47
(三) 血液處理……………………………………………………….47
(四) 血漿葡萄糖濃度之測定……………………………………….48
(五) 血漿胰島素濃度之測定……………………………………….48
(六) 血漿三酸甘油酯濃度之測定…………………………………48
(七) 血漿總膽固醇濃度之測定……………………………………49
(八) 犧牲時樣本收集與處理……………………………………….49
(九) 肝臟脂質萃取.............................................................................49
(十) 肝臟三酸甘油酯濃度之測定………………………………….50
(十一) 肝臟總膽固醇濃度之測定…………………………………….50
(十二) 肝臟 TBARS 濃度之測定........................................................50
第五節 統計分析……………………………………………………………….52
第五章 結果與討論…………………………………………………………………...58
第一節 實驗材料一般成分分析結果………………………………………….58
第二節 動物實驗結果………………………………………………………….59
實驗I、急性試驗………………………………………………………………59
一、 結果…………………………………………………………………….59
(一) 第一階段………………………………………………………...59
1. 急性試驗 (A) 正常鼠………………………………………..59
2. 急性試驗 (B) 代謝症候群鼠………………………………..60
3. 急性試驗 (C) 糖尿病鼠……………………………………..61
(二) 第二階段………………………………………………………...62
1. 急性試驗 (A) 正常鼠………………………………………..62
2. 急性試驗 (B) 代謝症候群鼠………………………………..62
3. 急性試驗 (C) 糖尿病鼠……………………………………..63
二、 討論…………………………………………………………………….63
實驗II、長效性試驗…………………………………………………………….65
一、 結果………………………………………………………………………66
(一) 高果糖動物模式……………………………………………….66
(二) 攝食量與生長狀況………………………………………...…..66
(三) 血壓變化……………………………………………………….66
(四) 空腹血糖值變化……………………………………………….66
(五) 第4週口服葡萄糖耐受性試驗………………………………..67
(六) 第8週口服葡萄糖耐受性試驗………………………………..67
(七) 第12週口服葡萄糖耐受性試驗………………………………67
(八) 組織臟器重量………………………………………………….68
(九) 血漿葡萄糖、胰島素、三酸甘油酯及總膽固醇之濃度變化.68
(十) 肝中三酸甘油酯、膽固醇及 TBARS 之濃度變化………….68
二、 討論………………………………………………………………………68
(一) 高果糖動物模式………………………………………………...68
(二) 攝食量與生長狀況……………………………………………...69
(三) 血糖與胰島素阻抗……………………………………………...69
(四) 高血壓…………………………………………………………...71
(五) 脂質代謝………………………………………………………...71
(六) 氧化壓力………………………………………………………...72
第六章 結論………………………………………………………………………...73
參考文獻……………………………………………………………………………….90
附錄………………………………………………………………………..………….101
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