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標題: | 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 Protective effects of ginseng essence on CCl4-induced oxidative stress and liver injury in rats |
作者: | Ching-Yi Weng 翁靜儀 |
指導教授: | 沈立言(Lee-Yan Sheen) |
關鍵字: | 人參精華液;四氯化碳;氧化壓力;護肝;人參皂苷, ginseng essence,carbon tetrachloride,oxidative stress,hepatoprotection,ginsenoside, |
出版年 : | 2013 |
學位: | 碩士 |
摘要: | 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. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17139 |
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