臺灣博碩士論文加值系統

運動的好處在近幾年不斷被提倡，但目前有許多研究證實，激烈或高強度運動不但會對身體不利，也會造成體內自由基過度增加，導致腎臟組織受到傷害。精胺酸 (L-arginine) 能增加肌肉生長及提升有氧運動的能力，並且具有減低氧化壓力與改善組織受損的效果。但部份文獻證實，在誘發腎小管結晶後，補充精胺酸反而會使腎臟疾病更加嚴重，甚至出現腎結石。故本篇研究目的有兩個：(一) 探討精胺酸是否能清除激烈運動所產生的自由基，進而降低腎臟的氧化壓力；(二) 觀察長期服用精胺酸是否會對腎臟造成負面的影響。實驗將 72隻 Spargue-Dawley 大鼠隨機分為靜態生活飲食控制組 (SC)、靜態生活補充精胺酸 (2 g/kg BW) (SC+Arg)、單次衰竭運動組 (E)、補充精胺酸合併單次衰竭運動組 (Arg+E)、單次衰竭運動後24小時犧牲組 (E-24) 及補充精胺酸合併單次衰竭運動後24小時犧牲組 (Arg+E-24h) 等六組，每組12隻。經30天餵養後，控制組大鼠立即犧牲，衰竭運動組大鼠在從事單次衰竭運動後，分成立即犧牲與24小時後犧牲。犧牲後收集血液及腎臟，檢測血液生化值、腎臟組織的黃嘌呤氧化酶 (xanthine oxidase，XO) 活性和TBARS 產量及抗氧化酵素的活性，包括 superoxide dismutase (SOD)、catalase、glutathione peroxidase 及 glutathione reductase。並以西方點墨法 (Western blot) 檢驗nitrotyrosine 蛋白表現。實驗數據都以SPSS 14.0 統計分析軟體套件進行單因子變異數分析 (One-Way ANOVA)，並以Tukey’s進行事後比較 (post hoc) 組間差異。每次實驗各組至少重複三次，數據以 Mean±SEM 表示，當p＜0.05 時，表示具有統計上之差異。結果顯示，血液生化值方面，衰竭運動顯著增加血漿尿素氮及運動 24 小時後的乳酸濃度，表示衰竭運動造成急性及延遲性的損傷。在抗氧化方面，精胺酸的補充能夠立即增加衰竭運動的過氧化氫酶活性，並將衰竭運動後24小時的過氧化氫酶活性維持在高活性，來降低延遲性的運動傷害。氧化壓力方面，精胺酸的補充確實能降低靜態生活、衰竭運動即衰竭運動後24小時XO的活性，也大大降低了運動所帶來的脂質過氧化的傷害。在nitrotyrosine 蛋白的表現上，補充精胺酸後，不論是靜態生活或是衰竭運動，都有顯著的增加。由本實驗的結果可知，補充精胺酸能夠有效地增加抗氧化酵素的活性，進而降低運動對腎臟所帶來氧化壓力。但若長期服用精胺酸，將會增加腎臟硝基酪胺酸蛋白增加而增加氮化壓力。

The advantage of physical exercise has been emphasized, however, strenuous exercise also resulted in overproduction of free radicals and leading to the damage of multiple tissues. L-arginine has been reported to promote the increasing of muscular growth and enhance the capability of aerobic exercise, and decrease oxidative stress as well as tissue damage. The previous studies indicated that supplementation of L-arginine could enhance the intrarenal oxidative stress in hyperoxaluric kidney with massive CaOx crystal deposition. Therefore, the aims of the study are to (1) study the effect of L-arginine supplementation on the reduction of exercise-induced free radical, (2) study whether the long-term supplementation of L-arginine can cause the negative impact in kidney. Seventy-two male Sprague-Dawley rats will be randomly assigned to six groups: (1) sedentary control, (2) sedentary control with 2% L-arginine supplementation, (3) exhaustive exercise, (4) 2% L-arginine supplementation and exhaustive exercise, (5) 24 h after exhaustive exercise, (6) 2% L-arginine supplementation and 24 h after exhaustive exercise. The animals will be kept in the control diets or L-arginine supplementation for 30 days. The exhaustive exercise groups will be performed with a single bout of strenuous exhaustive exercise on the treadmill. At the end of the study, the rats will be sacrificed as the experimental design. The production of TBARS, XO and the activities of antioxidant enzyme, including superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx) and glutathione reductase (GR) will be measured with commercial kits. Western blot will be performed to detect the protein expression of nitrotyrosine. All value were given as the mean±SEM. In some cases, the treatment means were tested for homogeneity by one-way ANOVA, and the different between specific means was tested for significance by Tukey’s test. A difference between two means was considered statistically significance when p<0.05. The results showed a significant increase in blood urea nitrogen and lactate concentration after exhaustive exercise. Supplementation of L-arginine attenuated the exhaustive exercise-induced increase of plasma creatinine and lactate concentration. Supplementation of L-arginine also increased the activities of catalase. On the oxidative stress, the activity of TBARS and XO were decreased by L-Arginine supplementation. But, long-term L-Arginine administration increased the nitrosative stress in kidney.

原創聲明書............................................... ii
學位考試審定書........................................... iii
中文摘要................................................. iv
英文摘要................................................. vi
謝誌..................................................... viii
目錄..................................................... ix
表目錄................................................... xii
圖目錄................................................... xiii
名詞縮寫................................................. xv

第壹章　緒論............................................. 1
第一節　研究背景...................................... 1
第二節　實驗假設...................................... 1
第三節　實驗目的...................................... 2
第貳章　文獻探討......................................... 3
第一節　腎臟生理...................................... 3
第二節　精胺酸...................................... 6
第三節　氧化壓力...................................... 11
第四節 精胺酸對氧化壓力的調節........................ 16
第參章　材料與方法....................................... 18
第一節　大鼠品種與飼養方法............................ 18
第二節　實驗設計...................................... 18
第三節　樣本分析項目與方............................. 20
第四節 資料處理與統計分析........................... 27
第肆章　實驗結果....................................... 29
第一節　體重及腎臟重量變化............................ 29
第二節　血液生化值.................................... 29
第三節 補充精胺酸對於衰竭運動誘發氧化壓.............. 31
第四節 補充精胺酸對大鼠腎臟中硝基酪胺酸蛋白表現的影響 33
第伍章　討論............................................. 34
第一節　補充精胺酸對衰竭運動大鼠血液生化值的影響...... 34
第二節　補充精胺酸對於衰竭運動後抗氧化的表現.......... 36
第三節 氧化酵素及脂質過氧化程度的表現............... 37
第四節 硝基酪胺酸蛋白的表現.......................... 38
第陸章 結論............................................ 39
參考文獻................................................ 40
中文部分............................................ 40
英文部分............................................ 40

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