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研究生:蔡韶恩
研究生(外文):Shao-En Tsai
論文名稱:多功能組合性保健食品配方之護肝效應
論文名稱(外文):Hepatoprotective effect of multi-functional and multi-ingredient health food formula
指導教授:顏國欽顏國欽引用關係
學位類別:碩士
校院名稱:國立中興大學
系所名稱:食品暨應用生物科技學系所
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
畢業學年度:97
語文別:中文
論文頁數:148
中文關鍵詞:保健食品氧化壓力抗氧化四氯化碳護肝
外文關鍵詞:health foodoxidative stressantioxidantcarbon tetrachloride and hepatoprotection
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近年來國人使用保健食品預防疾病的風氣日盛,而肝病為我國國病,故如何以保健食品之概念來預防肝病亦顯重要。因此,本研究選取多種具生理活性之食材,組成多功能組合性保健食品 I 及 K (簡稱配方 I 及 K),期能利用組合性的方式獲得比單方產品更佳之生理功效,進而開發出具有護肝功效之組合性保健食品。首先利用抗氧化分析及四氯化碳誘導大鼠慢性肝損傷實驗,評估配方 I 及 K 之護肝功效。結果顯示,配方 K 在抗氧化分析實驗及四氯化碳誘導大鼠慢性肝損傷實驗中,皆比配方 I 有較好的表現。綜合以上結果進一步修正配方並開發出多功能組合性保健食品 J (簡稱配方 J)。抗氧化分析結果顯示,配方 J 於 TEAC (1.9 mmol trolox equivalent/g formula) 及 ORAC (2.5 mmol trolox equivalent/g formula) 試驗中均具良好之抗氧化力。而由抗氧化物含量試驗結果顯示,配方 J具有豐富的總多酚 (153 mg gallic acid equivalent/g formula) 及類黃酮 (76 mg rutin equivalent/g formula)。更進一步以 HPLC 分析後發現配方 J 主要含有 chlorogenic acid、epicatechin 及 apigenin 等多酚及類黃酮化合物。動物護肝功效評估實驗方面,餵食大鼠四氯化碳 8 周並給予配方 J 中劑量 (0.3 g/kg bw) 組,其血清中 AST 活性可顯著降低 (p < 0.05)。由肝臟抗氧化酵素分析結果得知,餵食配方 J 0.1 g/kg bw (低劑量) 組即能顯著提昇 GRd 及 GST 之酵素活性。而經由組織病理切片 H&E 及 Masson''s 染色顯示,配方 J 0.9 g/kg (高劑量) 組能有效減輕肝臟中纖維化的生成。除此之外,經由細胞實驗證實,配方 J 對大鼠肝細胞 Clone 9沒有毒性,但對大鼠肝星狀細胞生長卻有明顯之抑制效果 (p < 0.05),配方 J 可能透過此方式而具有減緩肝纖維化之效果。綜合上述結果,推測配方 J 具有良好的抗氧化力,並能提高抗氧化素活性表現進而有效抑制由 CCl4 所誘導所產生之肝毒性,證實配方 J 具有減緩肝纖維化護肝之潛力。
In recent years, people have started to eat health foods to prevent different kinds of diseases. Liver diseases including chronic hepatitis, liver cirrhosis and hepatocellular carcinoma are common diseases in Taiwan. Therefore, maintenance a healthy liver is an important issue for the Taiwanese population. In this study, we choose several beneficial and functional herbs to develop multi-ingredient and multi-functional health food formulas I and K (formula I and K), and expected these complex formulas would better physiological activity than any single component. Further, we developed multi-ingredient and hepatoprotective health food. At first, we assessed hepatoprotective effects of formula I and K by analyzing antioxidant capacity and chronic liver damage induced by carbon tetrachloride (CCl4) in rats. We found formula K showed better efficiency than formula I. According to this result, we amended and developed the multi-ingredient and multi-functional health food formula J (formula J). Formula J showed potent antioxidant capacity by TEAC and ORAC. The TEAC and ORAC values of formula J were equal to 1.9 and 2.5 mmol trolox equivalent/g formula, respectively. The phenolics and flavonoids contents were equal to 153 mg gallic acid equivalent/g formula and 76 mg rutin equivalent/g formula. Furthermore, by HPLC-photo diode array (HPLC-PDA) analysis, we found chlorogenic acid, epicatechin and apigenin were the major phenolic and flavonoid compounds in formula J. In the animal study, rats were induced with CCl4 for 8 weeks we found that a supplement of middle-dose formula J (0.3 g/kg of bw) significantly (p < 0.05) reduced serum AST activity in rats induced with CCl4. When hepatic antioxidant enzymes activity was measured, low-dose formula J (0.1 g/kg of bw) significantly (p < 0.05) reversed glutathione reductase (GRd) and glutathione S-transferase (GST) activities. In liver histopathology H&E and Masson’s stains, high-dose formula J (0.9 g/kg of bw) reduced fatty liver and collagen accumulation. In cell culture experiment, we found formula J had no toxicity on rat hepatocyte Clone 9 cells. However, formula J could significantly inhibited the cell growth of rat liver stellate (HSC-T6) cell (p < 0.05), which may play a part in the effect of formula J in diminishing liver fibrosis. In conclusion, these data suggest that formula J has good increased antioxidant capacity and suppresses chronic liver damage induced by CCl4 through increasing activity of hepatic antioxidant enzymes and decreasing collagen accumulation. Therefore, formula J could be a potent hepatoprotective product.
目錄

全文摘要 (中) ……………………………………………… І
全文摘要 (英) ……………………………………………… III
目次………………………………………………………….. …. V
圖次……………………………………………………………… VIII
表次……………………………………………………………… V




前言…………………………………………………………………1
第一部分、文獻回顧 …………………………………………… 3
壹、肝臟的構造、功能與病變 ………………………………… 4
一、肝臟簡介……………………………………………………… 4
二、肝臟功能 …………………………………………………… 7
三、慢性肝疾病的成因 ………………………………………… 8
四、肝纖維化的形成機制 ……………………………………… 11
五、四氯化碳造成慢性肝損傷及肝纖維化之機轉……………… 18
貳、自由基與氧化傷害 ………………………………………… 21
一、自由基種類與來源………………………………………… 21
二、氧化傷害與疾病…………………………………………… 26
三、生物體內抗氧化防禦系統………………………………… 31
參、天然食材介紹 ……………………………………………… 34
一、綠茶………………………………………………………… 36
二、刺五加……………………………………………………… 37
三、金針菇……………………………………………………… 39
四、洋甘菊……………………………………………………… 40
肆、研究目的………………………………………………… 45
伍、研究架構…………………………………………………… 47
第二部分、多功能組合性保健食品配方 I 及 K 對四氯化碳大鼠誘導慢性肝損傷之保護效應………………… 49
中文摘要………………………………………………………… 50
英文摘要………………………………………………………… 51
前言……………………………………………………………… 53
實驗架構………………………………………………………… 54
材料與方法……………………………………………………… 55
結果與討論 …………………………………………………… 63
一、配方 I 及 K 抗氧化物含量分析…………………… 63
二、配方 I 及 K 抗氧化性分析………………………… 63
三、配方 I 及 K 之 HPLC 成分分析..………………… 64
四、體重與臟器重量變化………………………………… 65
五、血清生化值分析……………………………………… 66
六、抗氧化酵素分析……………………………………… 67
七、組織病理學分析……………………………………… 68
第三部分、多功能組合性保健食品配方 J 對四氯化碳誘導
鼠慢性肝損傷之保護效…………………………… 82
中文摘要………………………………………………………… 83
英文摘要………………………………………………………… 84
前言……………………………………………………………… 86
實驗架構………………………………………………………… 88
材料與方法……………………………………………………… 89
結果與討論……………………………………………………… 98
一、配方 J 之抗氧化成分及活性…………………………… 98
二、HPLC 成分分析…………………………………………… 98
三、血清生化值分析………………………………………… 99
四、體重與臟器重量變化…………………………………… 102
五、抗氧化酵素分析………………………………………… 103
六、組織病理學分析………………………………………… 105
七、明膠蛋白酶活性電泳分析法…………………………… 106
八、配方 J 對 TGF-β1 及 type І collagen 的 mRNA表現之影響 106
九、以配方 J培養星狀細胞及大鼠肝細胞 24 小時後之
之存活率影響…………………………………………… 107
十、以配方中所含 chlorogenic acid 或 apigenin 培養星狀細胞 24 小時後之對細胞生長之影響……………………… 108
十一、MMP-2 activity 電泳分析……………………………… 108
十二、胞內活性氧分析……………………………………… 108
總結論…………………………………………………………… 137
參考文獻………………………………………………………… 138
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