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研究生:翁靜儀
研究生(外文):Ching-Yi Weng
論文名稱:人參精華液於四氯化碳誘導大鼠肝損傷之動物模式中透過調節氧化壓力產生護肝功效
論文名稱(外文):Protective effects of ginseng essence on CCl4-induced oxidative stress and liver injury in rats
指導教授:沈立言沈立言引用關係
指導教授(外文):Lee-Yan Sheen
口試委員:邱智賢謝淑貞李宗貴
口試委員(外文):Chih-Hsien ChiuShu-Chen HsiehChong-Kuei Lii
口試日期:2013-07-02
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:2013
畢業學年度:101
語文別:中文
論文頁數:133
中文關鍵詞:人參精華液;四氯化碳;氧化壓力;護肝;人參皂苷
外文關鍵詞:ginseng essencecarbon tetrachlorideoxidative stresshepatoprotectionginsenoside
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肝臟是維持人體生命的重要器官,具有調節營養素的代謝、解毒及其他生化反應等功能。許多研究證實與生活習慣有關的文明病,如:慢性發炎、癌症等皆與活性氧屬 (Reactive oxygen species, ROS) 有關,因此降低自由基產生可能可以降低許多疾病的發生率。而人參精華液由四個主要的中草藥所組成:西洋參 (Panax quinquefolius)、人參 (Panax ginseng)、蓮子 (Nelumbo nucifera) 及百合 (Lilium longiflorum)。人參中最主要的活性成分是人參皂苷,許多研究顯示人參皂苷具有護肝、預防肥胖、預防腫瘤形成、預防糖尿病及減緩發炎等效果。
本研究旨在探討人參精華液樣品其有效成分人參皂苷 (ginsenosides) 含量,以及人參精華液對以四氯化碳誘導大鼠慢性肝纖維化實驗模式下的護肝功效評估及調節氧化壓力之效果。實驗使用雄性Wistar大鼠隨機分成6組:(A) 正常控制組;(B) 四氯化碳 (負控制) 組;(C) silymarin (正控制) 組 (0.5 g/kg bw/day);(D) 低劑量樣品組 (0.625 g/kg bw/day);(E) 中劑量樣品組 (1.250 g/kg bw/day);(F) 高劑量樣品組 (3.125 g/kg bw/day)。管餵 20% 四氯化碳 (1.5 ml/kg bw) 誘導大鼠產生慢性肝損傷,實驗期持續9週,包含8週的四氯化碳誘導期。探討其對於體內肝功能、氧化壓力及發炎反應相關評估指數之影響。
結果顯示,在活性成分分析方面,發現人參精華液含有人參皂苷Rg1、人參皂苷Re、人參皂苷Rb1、人參皂苷Rc、人參皂苷Rd和人參皂苷Rg3,且先前研究也指出,人參皂苷Rb1、Rg3及Rg1具有護肝功效。在動物實驗結果方面,隨飼養週數增加,各實驗組之平均體重皆有增加之趨勢,且相較於負控制組,人參精華液試驗樣品處理組有穩定的體重增加現象。經四氯化碳誘導後的負控制組臟器相對重量顯著高於控制組,在給予試驗樣品後,可以改善臟器發炎的情形。在肝功能評估方面,人參精華液可以有效地使血清中天門冬胺酸轉胺脢 (AST) 和丙胺酸轉胺酶 (ALT) 活性下降、血清白蛋白含量上升。另外,人參精華液亦可以降低肝臟中脂質,改善四氯化碳造成脂質代謝異常的現象。在組織病理切片分析結果中,亦顯示中給予人參精華液確實可以減緩肝損傷的程度。在氧化壓力評估方面,相較於四氯化碳處理組,餵食人參精華液也可以提高體內抗氧化酵素活性及抗氧化物質含量,並且減少肝臟中脂質過氧化物含量。在發炎反應評估方面,四氯化碳會引起肝毒性,造成發炎反應並促使發炎相關調控因子產生,給予人參精華液可以降低促發炎的細胞激素TNF-α、INF-γ、IL-6和TGF-β濃度,並抑制肝臟星狀細胞活化。
綜合上述動物實驗結果,推論人參精華液可以改善肝功能相關指標和脂質代謝異常的現象,並可以藉由提升體內抗氧化酵素活性和抗氧化物質含量,因而減緩氧化壓力和發炎反應,抑制星狀細胞活化而達到降低纖維化的情形。未來可望將人參精華液開發作為護肝保健食品。


Liver is a vital organ, which is responsible for the regulation of nutrients metabolism, detoxification functions and others biochemical reactions. Many reaserches have reported that civilization diseases, such as chronic inflammation, and cancer are related to reactive oxygen species (ROS). Therefore, reducing free radicals maybe decrease the incidence of many diseases.
The major components of ginseng essence include four herbs, Panax quinquefolius, Panax ginseng, Nelumbo nucifera and Lilium longiflorum. Ginsenosides are major active compounds in ginseng essence, and have been reported as the active compounds of hepatoprotection, anti-obesity, anti-tumor, anti-diabetic, and anti-inflammatory effects in many researches.
Therefore, the objective of this study is to evaluate the hepatoprotective effect of ginseng essence on carbon tetrachloride (CCl4)-induced liver fibrosis in animal model, investigate the potential mechanism in the regulation of oxidative stress and determine the contents of acive compounds (ginsenosides) in ginseng essence.
In the CCl4-induced animal model male Wistar rats were randomized into six groups: (A) control group was only treated with vehicle; (B) negative control group was treated with CCl4; (C) positive control group was treated with CCl4 and silymarin (0.5 g/kg bw/day); (D) low dose group was treated with CCl4 and 0.625 g/kg bw/day ginseng essence; (E) medium dose group was treated with CCl4 and 1.25 g/kg bw/day ginseng essence; (F) high dose group was treated with CCl4 and 3.125 g/kg bw/day ginseng essence. Using oral administration of 20% CCl4 (1.5 ml/kg bw) induced chronic liver damage in rats. This experiment was performed for 9 weeks, including 8-week induction of CCl4. The effects of ginseng essence on liver function, oxidative stress and inflammation asseeement index in vivo were investigated.
In the results of determinations of active compounds, there are ginsenoside Rg1, Re, Rb1, Rc, Rd and Rg3 in ginseng essence. Three of them, ginsenoside Rb1, Rg3 and Rg1 have been reported to exert hepatoprotective effect by previous studies. In the results of animal experiment, all ginseng essence treatments stablely increase body weight as compared to negative control group. After induction of CCl4, negative control group have higher relative organ weight than control group. However, after the oral administration of ginseng essence can improve the inflammation in organs. Regarding liver function, ginseng essence can effectively descrease the activities of AST and ALT, and increase albumin in serum. Additionaly, ginseng essence can also descrease lipids in liver, and improve CCl4-induced abnormal metabolism of lipid. In the results of histopathological analysis, the oral administration of ginseng essence can reduce the score of liver damage. In terms of oxidative stress, as compared to carbon tetrachloride group, the oral administration of ginseng essence can also elevate the activities of antioxidative enzymes and the conten of antioxidant, and reduce the content of lipid peroxides. Moreover, CCl4 can cause hepatotoxicity, and results in inflammation and promote the production of inflammatory mediators. The oral administration of ginseng essence can descrease the content of pro-inflammation cytokine, such like TNF-α、INF-γ、IL-6 and TGF-β, as well as inhibit the activation of hepatic stallate cells.
In conclusion, ginseng essence could ameliorate the oxidative stress and inflammation to improve liver function and abnormal metabolism of lipid in CCl4–induced rats. Additionally, it could inhibit the activation of hepatic stallate cells by elevating the activities of antioxidative enzymes and the content of antioxidant, and then can ameliorate liver fibrosis. Therefore, ginseng essence could be a promising hepatoprotective product in the future.


謝誌 Ⅰ
中文摘要 II
英文摘要 Ⅲ
目錄 Ⅴ
圖次 Ⅷ
表次 IX
第一章、前言 1
第二章、文獻回顧 3
第一節、肝臟之簡介 3
一、肝臟介紹 3
二、肝臟之生理功能 3
三、與肝臟相關之疾病 6
第二節、自由基之簡介 9
一、何謂自由基與活性氧 9
二、自由基與活性氧的形成 11
三、自由基和活性氧對於生物體的傷害 12
第三節、人體內的抗氧化防禦系統 13
一、抗氧化物 14
二、抗氧化酵素 15
第四節、四氯化碳 16
第五節、水飛薊素之護肝功效 18
第六節、本實驗使用之人參精華液複方簡介 19
一、西洋參 (Panax quinquefolius L.) 19
(一) 西洋參簡介 19
(二) 西洋參之生理活性 21
二、人參 (Panax ginseng C.A. Meyer) 23
(一) 人參簡介 23
(二) 人參之生理活性 24
(三) 人參中活性成分 25
三、百合 (Lilium longiflorum) 30
(一) 百合簡介 30
(二) 百合之生理活性 30
四、蓮子 (Nelumbo nucifera Gaertn) 31
(一) 蓮子簡介 31
(二) 蓮子之生理活性 32
第三章、實驗架構 35
第四章、材料與方法 36
第一節、實驗材料 36
一、實驗動物 36
二、樣品之來源 36
三、試藥 36
四、儀器設備 37
第二節、實驗方法 38
一、人參精華液食療方製備方法 38
二、人參精華液中有效成分分析 38
三、動物實驗設計 39
四、血清生化值檢測 41
五、肝臟中抗氧化能力測定 44
六、發炎反應相關細胞激素測定 50
七、組織病理切片觀察 52
八、免疫組織化學 52
九、統計分析 53
第五章、結果與討論 55
一、人參精華液中活性成分人參皂苷之分析 55
二、平均體重變化 55
三、血清中ALT及AST之結果 56
四、血清中白蛋白含量 57
五、肝臟、脾臟、和腎臟絕對及相對重量 58
六、血清和肝臟中TG及TC含量 60
七、體內抗氧化防禦系統 61
八、肝臟中TBARS之結果 63
九、血清中細胞激素TNF-α及IFN-γ之結果 64
十、血清中細胞激素IL-6及total TGF-β1之結果 66
十一、免疫組織化學染色 α-SMA之結果 67
十二、組織病理切片 68
第六章、結論 69
第七章、結果圖表 70
第八章、參考文獻 91

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