臺灣博碩士論文加值系統

肉蓯蓉 (Cistanche tubulosa) 為多年生寄生草本植物，分布於溫暖的沙漠、荒漠等地區，為傳統珍貴藥材並素有“沙漠人參”之美譽，研究指出肉蓯蓉具有抗氧化、對於肝及腦損傷皆有保護作用，但對於抗疲勞之功效尚未明瞭。本研究目的在探討肉蓯蓉對老化促進小鼠抗疲勞之功能評估。實驗以3月齡雄性老化促進小鼠SAMP8 (senescence accelerated mouse prone-8, SAMP8) 為對象，隨機分有跑步機訓練組和無跑步機訓練組，每組又分為控制組和實驗組：控制組 (sedentary control, SC)、實驗組 (sedentary treatment, ST) 運動組 (exercise control, ExC)、運動實驗組 (exercise treatment, ExT)，每組10隻，總共4組，實驗組管灌200 mg/kg BW肉蓯蓉萃取物 (Cistanche tubulosa Extract, CE)，實驗為期4週。實驗期間記錄體重、攝食量和飲水量，實驗第3週進行老化評估，且所有組別於第1、2和4週執行運動測試 (exercise program)，記錄小鼠運動期間因疲勞而被電擊次數，隨後犧牲。分析血中乳酸、葡萄糖含量，及一般血液生化值，檢測肌肉和肝臟肝醣含量以探討可能增加運動能力之機轉，觀察血液、肝臟和腦之抗氧化狀態，包括丙二醛含量、麩胱苷肽過氧化酶活性和血液超氧歧化酶活性。另以免疫組織化學染色觀察腦部β-類澱粉蛋白 (β-amyloid protein, Aβ) 沉積情形。研究結果顯示，各組體重、攝食量和飲水量均無顯著差異。老化評估方面，運動實驗組之老化總分顯著較其他三組低 (p < 0.05)。跑步機訓練方面，運動實驗組在實驗期間第1、2和4週其運動表現比其他三組好、被電擊次數最少；實驗組和運動實驗組運動後其血液乳酸含量顯著比控制組低 (p < 0.05)；在肝醣檢測方面，運動實驗組其肝臟和肌肉肝醣含量高於其他三組；在血液抗氧化方面，運動實驗組其丙二醛含量為三組最低者、而麩胱苷肽過氧化酶活性為三組最高者，特別是超氧歧化酶活性，實驗組和運動實驗組其活性皆顯著高於控制組和運動組 (p < 0.05)；在肝臟和腦之抗氧化狀態，實驗組和運動實驗組其丙二醛含量低於控制組和運動組，麩胱苷肽過氧化酶活性則是實驗組和運動實驗組較其他二組高，特別是腦部的麩胱苷肽過氧化酶活性，實驗組和運動實驗組顯著較控制組和運動組高 (p < 0.05)。在小鼠腦部β-類澱粉蛋白沉積百分比方面，控制組和運動組其β-類澱粉蛋白沉積較實驗組和運動實驗組高，其中運動實驗組之β-類澱粉蛋白沉積百分比顯著較控制組低 (p < 0.05)。由實驗結果推論，肉蓯蓉萃取物有助於跑步機運動表現，降低運動後血液代謝產物生成乳酸，提升肌肉和肝臟肝醣的儲存量以及改善體內抗氧化狀態，進而達到抗疲勞效果，另外在腦部β-類澱粉蛋白沉積中發現肉蓯蓉能有助於小鼠的學習記憶能力。

Cistanche tubulosa is perennial parasite herbs and mainly distributed in warm deserts and arid lands, which is a precious plant for traditional medicine and herbs earned the honor of “Ginseng of the deserts”. Experimental results have demonstrated that Cistanche tubulosa has been shown to have anti-oxidant, and protective effect of liver and brain damage. However, no reference was published for investigating the effect of anti-fatigue. The purpose of this study was to investigate the effects of Cistanche tubulosa on anti-fatigue in senescence-accelerated mice (SAMP8). 3-month-old male SAMP8 mice were used in this study. The mice were randomly divided into four groups, sedentary and exercise were administered by treadmill to control and treatment groups. Groups were divided: sedentary control (SC, n=10), sedentary treatment (ST, n=10), exercise control (ExC, n=10), and exercise treatment (ExT, n=10). The control groups were given double-distilled water (ddH2O) by gavage and the treatment groups were given 200 mg / kg body weight (BW) Cistanche tubulosa extract (CE) by gavage for 4 consecutive weeks. The body weight, food intakes and water consumption were measured. And on the third week, the aging scores were measured. The exercise program was performed after trained 1, 2 and 4 week by all groups, and during exercise that the electric shock frequencies of exercise program were record. The blood glucose and lactic acid concentrations and biochemical parameters of blood were analyzed after the mice sacrificed. And we explored the possible mechanisms in increasing the capability of exercise by glycogen content of muscle and liver. Anti-oxidation state were estimated in blood, liver and brain, malondialdehyde concentration (MDA), glutathione peroxidase actives (GPx) and superoxide dismutase actives (SOD) of blood, and β-amyloid protein (Aβ) deposition of brain were evaluated. The results showed that there were no significant difference in body weight, food intakes and water consumption among groups. The aging score of ExT group was significantly lower than other groups (p < 0.05). Results from the exercise program, during the experiment, at the first, second and fourth week of exercise program that retention frequencies of ExT group was the least. After running, blood lactic acid concentrations of ST and ExT groups were significantly lower than SC and ExC groups (p < 0.05). The content of hepatic and muscle glycogen of ExT group was significantly higher than other groups (p < 0.05). Results from anti-oxidation state of blood showed that the MDA concentration of ExT group was lower than other groups, and GPx activity of ExT group was higher than other groups, especially the SOD activity of ST and ExT groups were significantly higher than SC and ExC groups (p < 0.05). In addition, the anti-oxidation state of liver and brain showed that the MDA concentration of liver and brain in ST and ExT groups were lower than other groups, and the GPx activity of ST and ExT groups were higher than other groups, particularly the GPx activity of brain in ST and ExT groups were significantly higher than SC and ExC groups (p < 0.05). In addition, the β-amyloid protein deposition of brain in ExT group was significantly lower than SC group (p < 0.05). In summary, the supplement of Cistanche tubulosa extract could increase capability of exercise, decrease blood lactic acid after exercise, increase capability of exercise by increasing glycogen content in muscle and liver and improve the antioxidant defense system. There effects show that Cistanche tubulosa extract have anti-fatigue activity. In addition, β-amyloid protein deposition of brain showed Cistanche tubulosa extract was taken benefit of learning and memory ability.

目錄

中文摘要.......................................................................................................I
英文摘要.....................................................................................................III
誌謝.............................................................................................................VI
目錄............................................................................................................VII
表目錄..........................................................................................................X
圖目錄.........................................................................................................XI
第一章 前言.................................................................................................1
第二章 文獻回顧.........................................................................................4
第一節 肉蓯蓉.........................................................................................4
一、肉蓯蓉簡介.................................................................................4
二、肉蓯蓉的生理功能.....................................................................5
第二節 運動與疲勞.................................................................................8
第三節 運動與氧化壓力........................................................................11
一、運動氧化壓力來源....................................................................11
二、運動與體內抗氧化狀態............................................................14
三、運動與組織傷害 ......................................................................16
第四節 老化促進小鼠.............................................................................17
一、老化促進小鼠來源.....................................................................17
二、老化促進小鼠之特徵與應用.....................................................17
三、老化指數系統.............................................................................19
第三章 材料與方法.....................................................................................26
第一節 實驗動物.....................................................................................26
第二節 實驗材料.....................................................................................27
一、實驗分組.....................................................................................27
二、劑量換算.....................................................................................27
第三節 實驗方法與步驟.........................................................................29
一、實驗流程.....................................................................................29
二、跑步機運動.................................................................................32
三、老化指數.....................................................................................33
四、血液生化學分析.........................................................................33
五、血液乳酸分析.............................................................................34
六、肝臟肝醣和肌肉肝醣分析.........................................................34
七、抗氧化能力分析.........................................................................35
八、腦部病理切片觀察.....................................................................41
第四節 統計分析...................................................................................50
第四章 結果.................................................................................................51
第一節 體重變化和平均每日飲水量及攝食量.....................................51
第二節 運動電擊次數.............................................................................53
第三節 老化指數.....................................................................................58
第四節 器官重量.....................................................................................61
第五節 血液生化學分析.........................................................................61
第六節 血液葡萄糖和乳酸含量.............................................................64
第七節 肝醣含量.....................................................................................66
第八節 血液、肝臟和腦抗氧化能力分析.............................................68
一、血液抗氧化能力.........................................................................68
二、肝臟和腦抗氧化能力.................................................................68
第九節 腦部β-類澱粉蛋白之沉積........................................................75
第五章 討論.................................................................................................78
第六章 結論.................................................................................................85
第七章 參考文獻.........................................................................................86

表目錄

表一 SAM品系之病理特徵......................................................................................22
表二 SAM品系之病理表現與平均生命期..............................................................23
表三 老化指數判定標準..........................................................................................24
表四 3月齡雄性小鼠飼養4週之體重變化和平均每日飲水量及攝食量...........52
表五 3月齡雄性小鼠飼養3週後之老化指數比較...............................................60
表六 3月齡雄性小鼠飼養4週後之器官重量.......................................................62
表七 3月齡雄性小鼠飼養4週後之血液生化值比較...........................................63
表八 3月齡雄性小鼠飼養4週之跑步機測試後血液葡萄醣和乳酸含量...........65
表九 3月齡雄性小鼠在跑步機運動後之肝臟和肌肉肝醣含量...........................67
表十 3月齡雄性小鼠飼養4週後之血液丙二醛含量及抗氧化酵素麩胱苷肽過氧化酶和超氧歧化酶活性.70

圖目錄

圖一 SAM品系小鼠的世代發展.............................................................................21
圖二 實驗流程圖.....................................................................................................31
圖三 跑步機裝置圖.................................................................................................32
圖四 腦部Ｆ區域切面圖.......................................................................................48
圖五 病理組織切片流程圖...................................................................................49
圖六 (A) 3月齡雄性小鼠第1週運動測試之電擊次數......................................55
圖六 (B) 3月齡雄性小鼠第2週運動測試之電擊次數......................................56
圖六 (C) 3月齡雄性小鼠第4週運動測試之電擊次數......................................57
圖七 3月齡雄性小鼠飼養3週後之外觀...........................................................59
圖八 3月齡雄性小鼠飼養4週後之肝臟丙二醛含量.......................................71
圖九 3月齡雄性小鼠飼養4週後之腦部丙二醛含量.......................................72
圖十 3月齡雄性小鼠飼養4週後之肝臟麩胱苷肽過氧化酶活性...................73
圖十一 3月齡雄性小鼠飼養4週後之腦部麩胱苷肽過氧化酶活性...............74
圖十二 SAMP8小鼠腦部β-類澱粉蛋白沉積情形............................................76
圖十三 3月齡雄性小鼠飼養4週後之腦部β-類澱粉蛋白沉積平均面積百分比. 77

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