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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/14581

Title: 吳郭魚為動物模式評估一條根降血脂、降膽固醇功能及抗發炎與抗環境緊迫之功能
Development of tilapia as an animal model for evaluation of I-Tiao-Gung (Glycine tomentella) on hyperlipidemia, hypercholesterolemia, inflammation and environmental stress
Authors: Tsui-Yao Chen
陳翠瑤
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: 一條根;吳郭魚;抗發炎;降血脂;降膽固醇
I-Tiao-Gung, Glycine tomentella;tilapia;anti-inflammation;hypolipidemia;hypocholesterolemia
Date: 2005
Issue Date: 2011-06-30T07:40:04Z
Abstract: 摘 要 金門產闊葉大豆屬一條根 (Glycine tomentella Hayata) 民間用於治療風濕、關節炎、筋骨酸痛,但其藥理作用未明。本研究先以體外試驗 (in vitro) 探討一條根之抗氧化及抗發炎作用,再以吳郭魚飼養試驗 (in vivo),探討一條根之生理活性,建立以吳郭魚為模式動物初步篩選保健食品之功效。 一條根 95% 乙醇萃取物 (GT-E) 抑制亞麻油酸自氧化之 IC50 為 4.6 μg/mL,抗氧化力是水溶性抗氧化劑 trolox (IC50=14.2 μg/mL) 的 2.3 倍。清除 DPPH 的 EC50 為 18.4 μg/mL,與 trolox 相似 (17.1 μg/mL)。GT-E 抑制誘發型環氧合酶 (cyclooxygenase-2;COX-2) 的 IC50 為 42.0 μg/mL,較 indomethacin 高 (0.61 μg/mL),抑制 5-脂氧合酶 (5-lipoxygenase; 5-LOX) 的 IC50 為 0.40 μg/mL,較 nordihydroguaiaretic acid (NDGA) 的低 (2.3 μg/mL)。 以商用水產飼料 (蛋白質 53.0%,脂質 7.4%,灰分11.4%、粗纖維 1%,碳水化合物 21.4%,熱量 3194 kcal/kg) 餵食體重 185.0±19.3 g 雜交種吳郭魚 (Oreochromis niloticus × O. aureaus) 1.6 % body mass/day ,8 週後吳郭魚出現高血脂症及高膽固醇血症,血漿總三酸油酯為 1218.6±79.2 mg/dL,總膽固醇為740.3±276.3 mg/dL,低密度脂蛋白-膽固醇 (low density lipoprotein-cholesterol ; LDL-C) 392.1±56.3 mg/dL,另以相同條件飼養體重約 60 g 的莫三比克吳郭魚 (Oreochromis mossambicus) ,餵食 2% body mass/day 8 週後,血漿三酸甘油酯變為原來的 10.6倍,總膽固醇為 4.5 倍,LDL-C 為5.9 倍,與餵食 1% body mass/day 12 週後,均發生高血脂症及高膽固醇血症。 餵食體重 59.5± 9.5 g 雄性純種莫三比克吳郭魚 (Oreochromis mossambicus) 成魚,實驗組16尾,在飼料中添加1% GT-E 及控制組16 尾,餵食1.0% body mass/day 12 週後,GT-E 組較控制組之血漿三酸甘油酯低41.9%,膽固醇低30.4%、LDL-C也低 36.1%,均具有顯著差異 (p<0.05)。GT-E組血漿總抗氧化力 (total antioxidant status ; TAS) 顯著高於控制組 (p<0.05),其 LDL 粒子內的α-tocopherol 含量為 13.3±3.0 α-tocopherol molecule/LDL ,顯著高於控制組的 8.8±2.5 α-tocopherol molecule/LDL (p<0.05)。 以銅離子誘導LDL氧化之 in vitro 試驗,GT-E (3.2 μg/mL) 延長 LDL 氧化遲滯期的時間接近 trolox 0.5 μg/mL 的效果。離體 (ex vivo) 實驗, GT-E 組較控制組的 LDL 氧化遲滯期長,氧化速率較慢,其LDL 脂肪酸組成中,單元不飽和脂肪酸較控制組高、多元不飽和脂肪酸則較低,餵食 GT-E吳郭魚可保護 LDL 延緩氧化,並改變 LDL 脂肪酸的組成。 吳郭魚在氯化銨環境緊迫下血漿之 LOX 活性上升,攝食 GT-E 可抑制 LOX 活性。以 40 尾雄性雜交種吳郭魚成魚,分別餵食添加GT-E 飼料之實驗組 (20 尾)、與控制組 ( 20 尾) 12 個月後,曝露於 100 ppm 氯化銨水中,GT-E組與控制組的血比容積、血糖、AST、ALT 均無顯著變化,且兩組之間無顯著差異,但兩組的滲透壓在曝露氯化銨12 及 24 小時均較未曝露者低。在曝露至12 小時兩組吳郭魚的血小板5-LOX 活性明顯增加,控制組的活性顯著高於 GT-E 組 (p<0.05),其 12-LOX 及 15-LOX 活性也上升,GT-E 組的 12-LOX 及15-LOX則無明顯增加 (p>0.05),吳郭魚攝取GT-E 後在環境緊迫時,較能維持正常生理狀況。 整體而言,GT-E 可抑制 LDL 氧化、 LOXs 及COX-2 的活性,應具有抗發炎及抗動脈粥狀硬化的功效。GT-E 在ex vivo 及 iv vivo 實驗中,對高膽固醇血症及高血脂症的吳郭魚有降血脂、保護血漿中 LDL,延緩氧化的功效。由吳郭魚會發生高血脂症,其脂蛋白形態、脂肪酸代謝與膽固醇合成及利用類似哺乳動物,吳郭魚可做為評估降血脂的實驗動物,初步篩選具有降血脂功效的保健食品。
Abstract I-Tiao-Gung (Glycine tomentella Hayata) has been wildly cultured in Kinmen and used as a folk medicine for treatment of rheumatism, gout, arthritis, and ache of physique indicative of its anti-inflammatory activities. But its biological activities and pharmacological functions have not been studied systematically. The purpose of this study was to elucidate the long-claimed anti-inflammatory activity of I-Tiao-Gung by in vitro test and to assess its potential effects on atherosclerosis, using tilapia as an alternative animal to evaluate the in vivo effects of I-Tiao-Gung. The 95% ethanol extract of I-Tiao-Gung (GT-E) showed antioxidant activity on hemoglobin-catalyzed oxidation of linoleic acid. The IC50 value was 4.6 μg/mL lower than that of trolox (IC50=14.2 μg/mL). GT-E had the ablility to scavenge 1,1-diphenyl-2- picrylhydrazyl radicals (DPPH). Its EC50 value was 18.4 μg/mL similar to trolox (17.1 μg/mL). GT-E inhibited tilapia thrombocyte (the nucleated platelet) 5-, 12-, 15-lipoxygenase (LOX). The IC50 values were 0.43, 0.72 and 0.42 μg/mL respectively, while those of nordihydroguaiaretic acid (NDGA) on 5-, 12-, 15-LOX were 2.3, 1.6, and 1.7 μg/mL respectively. The IC50 on cyclooxygenase-2 (COX-2) inhibition by GT-E was 42.0 μg/mL while that for indomethacin (as a positive control) was 0.61 μg/mL. GT-E showed antioxidant activity by prolonging the lag phase (△Tlag) of human LDL oxidation induced by Cu2+ in a dose-dependent manner. The △Tlag of LDL added with 3.2 μg/mL GT-E was similar to that with trolox at 2.0 μM (ca. 0.5 μg/mL ). Hybrid tilapia (Oreochromis niloticus × O. aureaus), body mass 185.0±19.3, were fed daily with a commercial fish feed (protein 53.0%, lipid 7.4%, ash 11.4%, crude fiber 1%, carbohydrate 21.4%, total energy 3194 kcal/kg) at 1.6% body mass for 8 weeks. The plasma total triacylglycerol increased to 1218.6±79.2 mg/dL, total cholesterol increased to 740.3±276.3 mg/dL, and low density lipoprotein (LDL-C) increased to 392.1±56.3 mg/dL, respectively. Hyperlipidemia and hypercholesterolemia were found in fish. Mossambicus tilapia (Oreochromis mossambicus) (n=32), body mass 60±6.4 g, cultured with the same condition, fed daily 2% body mass for 8 weeks, and 1% body mass for 12 weeks were found hyperlipidemia and hypercholesterolemia. Matured male mossambicus tilapia (body mass 59.5±9.5, n=32) were randomly divided into two groups and fed with feed containing 1% (w/w) GT-E for 12 weeks. Total triacylglycerol of the GT-E group was lower than that of the control group by 41.9%, total cholesterol was lower by 30.4%, LDL-C was lower by 36.1%. Plasma total antioxidant status (TAS) of the GT-E group was higher than that of the control group by 79%. GT-E fed group had longer lag phase of Cu2+-induced LDL oxidation and retained significantly more α-tocopherol (13.3±3.0 α-tocopherol molecule/LDL) in LDL particles than the control group (8.8±2.5 α-tocopherol molecule/LDL). LDL from the GT-E group had more monounsaturated fatty acids and less polyunsaturated fatty acids than the control, indicating that GT-E may affect fatty acids metabolism. These evidences demonstrated that GT-E had hypolipidemic and hypocholesterolemic effects. Using tilapia exposed to ammonium chloride to evaluate the fuction of GT-E on anti-environmental stress. Hybrid male tilapia (n=40) weighed 185 g were randomly divided into two groups and fed with feed either containing 1% (w/w) GT-E or control diet for 12 months than exposed to 100 ppm ammonium chloride for up to 24 hours. Both groups decreased in osmosis measured at 12 and 24 hours. 5-LOX activity increased when exposed for 12 hours. Thrombocyte 12- and 15-LOX increased in the control group but not in the GT-E group. Intake of GT-E seemed to retard the changes in tilapia exposed to stress as abnormal concentration of ammonium chloride. The prevention of LDL oxidation and the dual inhibition of LOXs and COX-2 are indicative of the possible roles of GT-E on anti-atherosclerosis and anti-inflammation. Since GT-E inhibited LDL oxidation of tilapia in vitro, ex vivo and in vivo, GT-E had hypolipidemic and hypocholesterolemic effects in tilapia similar to that previously reported in hamster. Tilapia can serve as a surrogate animal model for prescreening hypocholesterol and anti-atherosclerosis effects of nutraceuticals and health foods.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0D90320004
http://ntour.ntou.edu.tw/ir/handle/987654321/14581
Appears in Collections:[食品科學系] 博碩士論文

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