臺灣博碩士論文加值系統

根據行政院衛生署2004年統計資料顯示，國人十大死因中，心臟病和腦血管疾病高居第二及第三位，依據流行病學研究報告指出，血脂異常及高血脂是導致心血管疾病的最大因素，此外，近年來已有許多研究指出活性氧及游離自由基會對人體健康造成傷害，甚至引發疾病，例如癌症或老化等。本研究目的係評估鹿角靈芝多醣體抗氧化活性與安全性及其調節血脂之功效。在鹿角靈芝多醣體性質測定部份，以β-1,3-葡聚醣含量的測定為指標，其含量以Laminarin equivalent(LE)表示，結果顯示每克子實體可萃取的粗多醣含量為10.03 mg，而其中所測得的β-1,3-葡聚醣為粗多醣之19.78%。抗氧化活性方面，利用清除DPPH自由基之能力、螯合亞鐵離子之能力、清除超氧陰離子的能力、還原力及硫氰酸鐵法評估，並以熱水及甲醇萃取物相比較，結果顯示在清除DPPH自由基能力及螯合亞鐵離子能力方面，以甲醇萃取物和多醣體效果最顯著；在清除超氧陰離子能力中，以多醣體效果最佳；然而在還原力及硫氰酸鐵法評估其抗氧化力之結果中，無論是熱水、甲醇萃取物及多醣體，皆隨著濃度的增加有上升的趨勢。藉由安氏試驗進行安全性評估，結果顯示，鹿角靈芝多醣體濃度為3、15、30 mg/ml時，不具有毒性及致突變性，且具有抗致突變性。動物實驗部分，在高油脂、高膽固醇的飲食條件下，經餵飼鹿角靈芝多醣體12週後，可藉由降低血漿總膽固醇及三酸甘油脂的濃度，達到調節血脂的功效，於病理組織切片的結果中，發現餵飼多醣體可達到抑制肝臟發生急性腫脹、壞死性肝炎及其他病灶的功效。

Based on the annual report of Department of Health ROC, heart disease and cardiovascular were ranked on second and third among ten leading causes of death in Taiwan 2004. According to epidemiologic studies, the hyperlipidemia and dyslipidemia are the greatest factors which cause cardiovascular disease (CHD). Recently research on the role of active oxygen and free radicals in tissue damage in various human diseases, cancer and aging are becoming increasingly recognized. The major objective of the current study was asessment of antioxidant activity and safety of Ganoderma amboinense and its hyperlipidemia effects on hamster. To
quantitatively determined the β-1,3-glucans isolated from Ganoderma amboinense and the results was presented as Laminarin equivalent (LE). The contents of extracted crude polysaccharides were 10.03 mg per gram of fruiting body and the extracted crude polysaccharides contained 19.78% of β-1,3-glucans. To elucidate antioxidant activity by scavenging effect on 1,1-dipheny-2-picryhydrazyl (DPPH) radical, chelating ability on ferrous ions, scavenging superoxide anion activity, ferric thiocyanate method and reducing power were well conducted. Results showed that methanol extract and polysaccharides illustrated the best effect on scavenging effect on 1,1-dipheny-2-picryhydrazyl (DPPH) radical and chelating ability on ferrous ions. On scavenging superoxide anion activity, polysaccharides demonstrated the highest effect. For antioxidant activity measured by the ferric thiocyanate method and reducing power of methanol, hot water extracts and polysaccharides increased with increasing amount of sample. Three different dosage of polysaccharides, 3, 15 and 30 mg/plate, depicted no toxicity and mutagenicity under specified conditions. Furthermore, the polysaccharides extracts from Ganoderma amboinense. indicated the best antimutagenic effect. In animal study, the results illustrated that intake polysaccharides can lower serum total cholesterol and triacylglycerol with a high fat and high cholesterol diet after 12 weeks experimental diet period. Histopathological changes of the liver, swelling, hepatitis and other symptom, were observed and can be
inhibited by feeding polysaccharides.

目錄

摘要 I
Abstract II
誌謝 IV
目錄 V
圖表目錄 IX
第1章 前言 1
第2章 文獻回顧 3
2.1 靈芝 3
2.1.1 生長與栽培 5
2.1.2 分類與特徵 6
2.1.3 活性成分 7
2.1.4 功效 8
2.2 抗氧化 12
2.2.1 氧化機制 12
2.2.2 天然抗氧化物 14
2.2.3 含抗氧化的活性成分物質 18
2.3 飲食與癌症 19
2.3.1 抗致突變劑的作用機制 19
2.3.2 食品中的致突變物 21
2.3.3 食物中的抗致突變物 24
2.3.4 安氏試驗 26
2.4 血脂 29
2.4.1 血脂分類與形成 29
2.4.2 天然調節血脂物質 38
第3章 材料與方法 40
3.1 實驗設計 40
3.2 實驗藥品 41
3.2.1 鹿角靈芝來源 41
3.2.2 一般化學藥品 41

3.2.3 多醣性質測定藥品 41
3.2.4 抗氧化藥品 41
3.2.5 安氏試驗藥品 42
3.2.6 動物試驗藥品 42
3.3 儀器設備 42
3.3.1 一般儀器 42
3.3.2 微生物試驗設備 43
3.3.3 調節血脂試驗設備 43
3.4 實驗方法 43
3.4.1 一般成分之分析 43
3.4.1.1 水分之測定 43
3.4.1.2 灰分之測定 43
3.4.1.3 粗脂肪之測定 44
3.4.1.4 蛋白質之測定 44
3.4.1.5 碳水化合物之測定 45
3.4.2 多醣性質之測定 45
3.4.2.1 粗多醣及總醣量的檢測 45
3.4.2.2 鹿角靈芝所含的β-1,3-D-葡聚醣檢測 46
3.4.3 抗氧化活性之測定 46
3.4.3.1 鹿角靈芝甲醇萃取物之製備 46
3.4.3.2 鹿角靈芝熱水萃取物之製備 47
3.4.3.3 清除DPPH自由基能力之測定 47
3.4.3.4 螯合亞鐵離子能力之測定 47
3.4.3.5 清除超氧陰離子之能力測定 48
3.4.3.6 還原力之測定 48
3.4.3.7 以硫氰酸鐵法測定之抗氧化力 48
3.4.4 安氏試驗 49
3.4.4.1 試驗菌株之活化 49
3.4.4.2 試驗菌株之保存 49
3.4.4.3 基因型態之確認 49
3.4.4.4 溶液之配製 50

3.4.4.5 毒性試驗 53
3.4.4.6 致突變性實驗 54
3.4.4.7 抗致突變性試驗 54
3.4.5 調節血脂動物試驗 55
3.4.5.1 攝取量之換算 55
3.4.5.2 飼料組成與製備 55
3.4.5.3 實驗動物與分組 55
3.4.5.4 樣品收集 57
3.4.6 血脂質分析 57
3.4.6.1 血漿脂蛋白的分離 59
3.4.6.2 血漿總膽固醇之測定 59
3.4.6.3 血漿三酸甘油酯之測定 59
3.4.6.4 血漿高密度脂蛋白膽固醇之測定 60
3.4.6.5 血漿低密度脂蛋白膽固醇之測定 60
3.4.6.6 肝臟、心臟及腎臟病理組織切片製作 60
3.4.7 統計分析 60
第4章 結果與討論 61
4.1 鹿角靈芝之一般成分分析 61
4.2 多醣性質之測定 64
4.2.1 粗多醣及總醣量的檢測 64
4.2.2 鹿角靈芝所含的β-1,3-D-葡聚醣檢測 64
4.3 鹿角靈芝抗氧化活性之探討 65
4.3.1 鹿角靈芝甲醇、熱水萃取液及多醣體之萃取率 65
4.3.2 清除 DPPH 自由基之能力 67
4.3.3 螯合亞鐵離子之能力 70
4.3.4 清除超氧陰離子之能力 72
4.3.5 還原力 74
4.3.6 以硫氰酸鐵法測定之抗氧化力 76
4.4 安氏試驗 78
4.4.1 試驗菌株基因型態之確認 78
4.4.2 毒性試驗 80

4.4.3 致突變性試驗 83
4.4.4 抗致突變性試驗 86
4.5 鹿角靈芝多醣體對倉鼠調節血脂功效評估 86
4.5.1 體重變化與飼料攝取量 86
4.5.2 血脂質分析 90
4.5.2.1 血漿總膽固醇(TC)濃度 90
4.5.2.2 血漿三酸甘油酯(TG)濃度 93
4.5.2.3 血漿高密度脂蛋白膽固醇(HDL-C)濃度 95
4.5.2.4 血漿低密度脂蛋白膽固醇(LDL-C)濃度 97
4.5.3 心臟、腎臟、肝臟及盲腸重量對於體重之值 99
4.5.4 外觀觀察及紀錄 99
4.5.5 組織切片 99
第5章 結論 109
參考文獻 110
作者簡介 135

















圖表目錄

圖 1、 抗致突變物的分類.........……………………………………........ 20
圖 2、 烹煮肉類食品中所分離之異環芳香胺類結構式………….…… 23
圖 3、 冠狀動脈的機制.......................………………………………….. 34
圖 4、 低密度脂蛋白代謝途徑……………………….………………… 35
圖 5、 鹿角靈芝子實體與粗多醣.……………………….……………... 62
圖 6、 鹿角靈芝萃取液清除DPPH自由基之能力……………………... 69
圖 7、 鹿角靈芝萃取液螯合亞鐵離子之能力...……………………….. 71
圖 8、 鹿角靈芝萃取液清除超氧陰離子之能力...…………………….. 73
圖 9、 鹿角靈芝萃取液之還原力.....…….……………………………... 75
圖10、 鹿角靈芝萃取液以硫氰酸鐵法之抗氧化力……………………. 77
圖11、 各組倉鼠於實驗期間血漿總膽固醇濃度變化..………………... 92
圖12、 各組倉鼠於實驗期間三酸甘油脂濃度變化……………………. 94
圖13、 各組倉鼠於實驗期間血漿高密度脂蛋白濃度變化......………... 96
圖14、 各組倉鼠於實驗期間血漿低密度脂蛋白濃度變化......………... 98
圖15、 各組心臟外觀之觀察.…………………........................................ 102
圖16、 各組腎臟外觀之觀察……………................................................. 103
圖17、 各組肝臟外觀之觀察....……………………………..................... 104
圖18、 各組心臟組織病理學鏡檢............................................................. 105
圖19、 各組腎臟組織病理學鏡檢......………………............................... 107
圖20、 各組肝臟組織病理學鏡檢....……………………………………. 108

表1、 活性氧和相關化合物之例……….…………………………….... 13
表2、 自由基之例…………………….......…………………….............. 15
表3、 抗氧化活性成分……………………............................................. 16
表4、 食物中的化學防護劑…………………......................................... 25
表5、 常見之基因型態….……………………………………………… 27

表6、 十大死因…………………………………………………............. 30
表7、 心血管疾病危險因子…………………………………................. 31
表8、 脂蛋白分類…………………………………................................. 32
表9、 低密度脂蛋白膽固醇、總膽固醇及高密度脂蛋白膽固醇的分級………………………………………......................................... 36
表10、 調節血脂的藥物…………..…………….……………………… 37
表11、 基礎飼料成分……………………………………………………. 56
表12、 動物實驗分組................................................................................. 58
表13、 鹿角靈芝子實體之一般組成......................................................... 63
表14、 鹿角靈芝熱水萃取物之區分......................................................... 66
表15、 靈芝甲醇、熱水萃取液及多醣體之萃取率................................... 68
表16、 試驗菌株S. typhimurium TA 98及TA 100之基因形態............... 79
表17、 鹿角靈芝多醣體對S. typhimurium TA98之毒性試驗.................. 81
表18、 鹿角靈芝多醣體對S. typhimurium TA100之毒性試驗................ 82
表19、 鹿角靈芝多醣體對S. typhimurium TA98之致突變性試驗.......... 84
表20、 鹿角靈芝多醣體對S. typhimurium TA100之致突變性試驗........ 85
表21、 鹿角靈芝多醣體對S. typhimurium TA98之抗致突變性試驗...... 87
表22、 鹿角靈芝多醣體對S. typhimurium TA100之抗致突變性試驗.... 88
表23、 不同飲食之倉鼠於實驗飲食前、實驗飲食十二週後之起始體重、最終體重及每日體重增加量.................................................. 89
表24、 不同飲食之倉鼠於實驗飲食前、實驗飲食十二週後之起始飼
料攝取量、最終飼料攝取量及攝食增加量................................... 91
表25、 不同飲食之倉鼠於實驗飲食十二週後之心臟、腎臟、肝臟及
盲腸重量相對於體重之百分比..................................................... 100
表26、 不同飲食之倉鼠於實驗飲食十二週後之肝臟、心臟、腎臟之
病理組織切片記錄......................................................................... 101

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