臺灣博碩士論文加值系統

薔薇紅景天(Rhodiola rosea)，生長在東歐及亞洲海拔4000公尺以上的高寒環境，屬於珍貴食用植物為我國傳統的藥用植物之一，在中醫古籍醫典中均有詳載和介紹是目前廣為研究的主題之一。本研究針對薔薇紅景天萃取物之抗氧化及抗疲勞功效，先利用高效液相層析質譜儀(HPLC-MS)將其中之特殊成份定量，接著探討薔薇紅景天特有單一成份及複方成份萃取物之抗氧化能力。此外，研究從動物實驗模式中觀察給予大鼠補充薔薇紅景天萃取物飲用水四週後，是否降低大鼠游泳運動後體內之氧化傷害、提升抗氧化能力、增加大鼠運動能力及改善疲勞情形。抗氧化功效之檢測是以化學冷光分析方法，分別觀察單一成份及複方成份萃取物降低O2.-、H2O2、HOCl自由基產生之能力。動物實驗則以雌性 wistar rat (180~200g)，分別餵食含有低(5mg/day/rat)、中(25mg/day/rat)、高(125mg/day/rat)三種劑量之含薔薇紅景天萃取物飲用水四週後給予負重游泳測試，觀察補充薔薇紅景天萃取物是否增加游泳時間改善其運動能力；另以不負重游泳九十分鐘模式，觀察肝臟之抗氧化酵素，包括Cu/Zn SOD、Mn SOD、CAT(Catalase)等酵素之蛋白質表現量、血液及肌肉中脂質過氧化物丙二醛（MDA）濃渡及血液中代謝產物包括：尿素氮（BUN）、肌酸酐（Creatinine）、乳酸（Lactic acid）等之產生量，並且探討可能增加運動能力之機轉，例如：能量代謝途徑之改變、增加肝臟及肌肉之肝醣儲存量、脂肪合成相關酵素(SREBP-1、FAS)之蛋白質表現量、提升體內組織含氧能力或肌肉纖維種類之改變。研究結果顯示薔薇紅景天不論其單一成份或複方成份萃取物，皆有顯著降低O2.-、H2O2、HOCl自由基產生之能力。動物實驗方面，大鼠補充含薔薇紅景天萃取物飲用水四週後，低、中、高劑量組別游泳時間皆顯著高於對照組，分別增加20.9%、49.6%、65%；不負重游泳運動後與對照組相互比較，飲用含有中、高劑量薔薇紅景天萃取物組別之大鼠，游泳後血液及肌肉中MDA濃度顯著降低、血液中之BUN及Creatinine亦有顯著降低的情形；飲用含中、高劑量薔薇紅景天萃取物組別大鼠之抗氧化酵素Cu/Zn SOD、Mn SOD蛋白質表現量上升；低、高劑量組別CAT酵素蛋白質表現量上升；飲用含中、高劑量薔薇紅景天萃取物組別之未游泳大鼠肝臟肝醣儲存量較高；飲用含薔薇紅景天萃取物組別大鼠之SREBP-1、高劑量薔薇紅景天萃取物組別大鼠之FAS酵素蛋白質表現量皆增加。大鼠靜脈注射薔薇紅景天萃取物(0.05mg/30min、0.1mg/30min)後，肝臟及肌肉組織含氧量上升，但在肌肉纖維種類表現方面，腿部肌肉的紅肌與白肌比例並沒有改變。研究結果推論：薔薇紅景天萃取物具有抗氧化功效、可提升大鼠體內之氧化酵素之蛋白質表現量、降低運動引起之氧化壓力且可能經由增加肝臟肝醣之儲存量、組織含氧量及脂肪合成相關之蛋白質增加運動能力、並且可能藉由能量利用之途徑改變，降低血液中代謝產物之生成量，改善運動後疲勞產生。

Abstract
The Rhodiola rosea, which grows in high altitude and cold environment in Eastern European and the Asian, is a precious edible plant for tradition medicine. However, the effect and mechanism on exercise induced fatigue and oxidative stress is uncertain. In this study, we evaluated the possible antioxidant activities of the extracts of Rhodiola rosea on exercise-induced oxidative stress and fatigue in the rats.
First, we identified the special ingredients of Rhodiola rosea by high performance liquid chromatography-mass spectrometer and evaluated the antioxidant activities of the extracts of Rhodiola rosea. In in vivo experiment, we assessed the effect of Rhodiola rosea extracts on exercise induced oxidative stress and fatigue. We fed three doses of Rhodiola rosea extracts in drinking water (5mg, 25mg, 125mg/day/rat) for four weeks in the female Wistar rats. We compared the capability of exercise, reactive oxygen species (ROS) response in blood, liver, skeletal muscle and kidney, the level of peroxidation products malondialdehyde (MDA) in serum and skeletal muscle and the biomarker of fatigue, including blood urea nitrogen (BUN), lactic acid, creatinine, creatinine kinase (CK), lactate dehydrogenase (LDH) and exercised-related enzymes by western blot in the rats undergoing 90- min swimming. We also explored the possible mechanisms in increasing the capability of exercise, like glycogen storage in liver and muscle, the activity of fat metabolism enzymes (SREBP-1 and FAS), the oxygen content in tissue or percentage changes of the muscle fiber types.
Our results showed that Rhodiola rosea extracts have a stronger scavenging activity against O2.-, H2O2 and HOCl. Treatment of Rhodiola rosea extracts in drinking water for four weeks increased the swimming time in a dose-dependent manner (20.9% at 5 mg, 49.6% at 25 mg and 65% at 125 mg). After the swimming test, the serum and muscle MDA concentrations and serum BUN and creatinine level were significantly decreased by Rhodiola rosea extracts treatment when compare to the control group. The expression in the antioxidant proteins like CuZn-superoxide dismutase, Mn-superoxide dismutase, catalase, heat shock protein 70, and Bcl-2 protein were all enhanced and the proapoptotic Bax protein was decreased after Rhodiola rosea treatment. Rhodiola rosea extracts treatment also increased liver glycogen content before swimming. Rhodiola rosea extracts increased SREBP-1 and FAS enzyme expression. After rodents injected vein with Rhodiola rosea extracts, the, the oxygen content show increase, but the ratio of type A and type B muscle never change. In conclusion: Rhodiola rosea extracts with antioxidant activities could enhanced the antioxidant proteins expression, decrease oxidative stress induced by exercise, and increase capability of exercise by increasing glycogen content in liver, increasing the oxygen content in tissues, and by enhancing the activity of fat metabolism enzymes. These effects can consequently decrease the levels of biomarker of fatigue and improve exercise-induced fatigue.

薔薇紅景天改善運動引起之氧化壓力及疲勞
論文口試委員省定書
博碩士論文電子檔案上網授權書
致謝
目錄
表次
圖次
中文摘要………………………………………………………………………………1
英文摘要……………………………………………………………………………．3
第壹章 前言…………………………………………………………………………5
第一節 研究動機…………………………………………………………………．5
第二節 研究目的………………….………………………………………………6
第貳章 文獻探討………………….………………………………………………7
第一節 自由基之介紹………………….…………………………………………7
第二節 運動與氧化壓力……………….…………………………………………12
第三節 運動與疲勞……………….………………………………………………14
第四節 骨骼肌纖維種類………………………………………………………….15
第五節 薔薇紅景天之抗氧化功效…………………………………………………16
第六節 細胞凋亡相關基因…………………………………………………………18
第七節 熱休克蛋白質………………………………………………………………18
第八節 脂肪合成相關酵素…………………………………………………………18
第參章 材料與方法…………………………………………………………………20
第一節 研究設計與架構……………………………………………………………20
第二節 研究材料與方法……………………………………………………………20
第三節 儀器設備……………………………………………………………………33
第四節 實驗藥品及來源……………………………………………………………34
第五節 統計分析……………………………………………………………………34
第肆章 結果…………………………………………………………………………35
第一節 薔薇紅景天之成份分析……………………………………………………50
第二節 薔薇紅景天之抗氧化功效…………………………………………………50
第三節 薔薇紅景天對運動能力之影響……………………………………………53
第四節 薔薇紅景天對疲勞改善之效果……………………………………………55
第伍章 討論…………………………………………………………………………72
第陸章 結論…………………………………………………………………………75
參考文獻………………………………………………………………………………76

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