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研究生:鍾宛儒
研究生(外文):Chung Wen-Ju
論文名稱:補充精胺酸對於大鼠衰竭運動後心臟損傷的影響
論文名稱(外文):Effect of L-arginine Supplementation on Exhaustive Exercise-Induced Cardiac Injury
指導教授:郭家芬郭家芬引用關係蔡秀純蔡秀純引用關係
指導教授(外文):Fannie C.F. KuoShiow-Chwen Tsai
學位類別:碩士
校院名稱:實踐大學
系所名稱:食品營養與保健生技學系碩士班
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2010
畢業學年度:98
語文別:中文
論文頁數:86
中文關鍵詞:精胺酸衰竭運動抗氧化酵素細胞凋亡
外文關鍵詞:L-arginineantioxidantexhaustive exerciseenzymesapoptosis
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近年來運動的好處一直不斷的被提倡,但目前有許多研究者證實,激烈或過度運動訓練不但對身體不利,也會造成體內自由基過度增加,導致心肌組織受到傷害。先前有文獻指出,精胺酸 (L-arginine) 能促進生長激素的分泌,提升肌肉生長及增強肌力,還具有減低氧化壓力與改善組織損傷等效果,但對應用於運動科學之研究,目前機制仍未清楚。因此本實驗目的有二: (一) 為探討精胺酸是否能清除激烈運動後所產生的大量自由基,進而降低心臟的氧化傷害與細胞凋亡; (二) 觀察補充精胺酸是否能幫助運動後恢復期的心臟復原,並分析可能機制。實驗將 72 隻 Sprague-Dawley 大白鼠隨機分為靜態生活飲食控制組 (SC)、靜態生活補充精胺酸組 (2 g/kg BW) (SC+Arg) 及運動組,再將運動組分為衰竭運動組 (E)、衰竭運動補充精胺酸組 (E+Arg) 、衰竭運動後 24 小時組 (E24) 及衰竭運動後 24 小時補充精胺酸組 (E24+Arg)。經 30 天餵養後,控制組大白鼠立即犧牲,衰竭運動組大鼠在跑步機從事單次急性衰竭運動後分成立即犧牲和 24 小時後犧牲。結果顯示,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。另外,精胺酸能夠立即增加運動後過氧化氫酶的活性與運動後24 小時超氧化歧化酶活性及榖胱甘肽和氧化型榖胱甘肽的比值。蛋白表現方面,發現衰竭運動及精胺酸的補充均會增加Bcl-2 及 connexin-43 蛋白的表現。心肌損傷方面,發現衰竭運動組心臟組織中 collagen 的分佈都有明顯的增加;除此之外,衰竭運動及衰竭運動後 24 小時均有 DNA 斷裂片段的產生,並且利用螢光染色觀察到同樣具有細胞凋亡的反應。精胺酸的補充可以減少心臟組織中 collagen 的分佈、DNA 斷裂片段以及心臟組織凋亡的情形,以達到減少衰竭運動對心臟立即性和延遲性的傷害。由本研究結果可知,補充精胺酸可能藉由增加抗氧化酵素的活化及減少細胞凋亡的產生,幫助運動後恢復期的心臟復原,補充精胺酸未來應可作為保護過度運動訓練所衍生之心臟傷害。
The advantage of physical exercise has been emphasized; however, strenuous exercise results in overproduction of free radicals, leading to the damage of cardiac tissues. L-arginine has been reported to promote the secretion of growth hormone, increase muscular strength, and decrease oxidative stress as well as tissue damages. To elucidate the effect of L-arginine supplementation on exercise, this experiment aims to (1) study the effect of L-arginine supplementation on the reduction of oxidative stress as well as apotosis during exhaustive exercise; (2) study the effect of L-arginine supplementation on the recovery of heart after exhaustive exercise. Seventy-two male Sprague-Dawley rats were randomly assigned to one of the six groups: (1) sedentary control; (2) sedentary control with 2% L-arginine supplementation; (3) exhaustive exercise; (4) exhaustive exercise with 2% L-arginine supplementation; (5) 24 h after exhaustive exercise; (6) 24 h after exhaustive exercise with 2% L-arginine supplementation. The animals were kept in the diets for 30 days. At the end of the study, the rats in control groups were sacrificed, the animals in exhaustive exercise groups were sacrificed right after performing a single bout of strenuous exhaustion exercise on a motorized treadmill, and the animals in “24 h after exhaustive exercise” groups were sacrificed 24 h after performing a single bout of strenuous exhaustion exercise. 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, superoxide dismutase, and GSH/GSSG ratio in animals of exercise groups. The expression of Bcl-2 and Connexin-43 were both enhanced in exercise groups with or without supplementation of L-arginine. The cardiac collagen distribution, DNA ladder, and TUNEL stain were significantly increased in the rats which performed exhaustive exercise; however, these damages were attenuated by L-arginine supplementation. The findings imply that L-arginine may apply in the protection of heart from damages caused by strenuous exercise.
第壹章 文獻回顧 15
第一節 自由基 15
一、自由基的定義 15
二、自由基的產生與清除 15
三、自由基對生理功能的影響 19
四、自由基誘發細胞凋亡的相關機制 21
第二節、運動 22
一、運動的生理反應 22
二、衰竭運動 24
三、運動與自由基 25
四、運動誘發心肌凋亡的相關機制 26
第三節 精胺酸 27
一、簡介精胺酸 27
二、精胺酸對運動表現的影響 27
三、精胺酸降低自由基的機制 28
四、精胺酸調節細胞凋亡的機制 29
第貳章 研究目的 30
第參章 實驗流程 31
第肆章 材料與方法 32
第一節 動物實驗設計 32
一、動物飼料組成與配置 32
二、動物分組與飼養 32
三、大白鼠適應與單次激烈衰竭運動之操作處方 33
第二節 動物犧牲與組織取樣 35
一、心肌細胞質中蛋白質萃取步驟 35
二、蛋白質含量之分析 36
三、抗氧化酵素檢測 36
四、西方點墨法 38
五、Masson’s trichrome stain 40
六、DNA片段電泳分析 40
七、TUNEL stain 41
第三節 統計分析 41
第伍章 實驗結果 43
第一節、體重及心臟重量變化 43
一、補充精胺酸對於大白鼠體重、心臟重及其比值之影響。 43
第二節、補充精胺酸對跑步適應前後心臟收縮功能的影響 45
一、補充精胺酸對心臟 fraction shortening 的影響 45
二、補充精胺酸對心臟心搏量的影響 45
三、補充精胺酸對心臟射出分率的影響 45
第三節、血液生化值 49
一、血糖濃度 49
二、血乳酸濃度 49
三、血漿肌酐酸激酶濃度 49
四、血漿尿素氮濃度 49
五、血漿尿酸濃度 49
六、血漿三酸甘油酯、肌酸酐、丙胺酸轉胺酶濃度 50
第四節、補充精胺酸對於心臟抗氧化能力的表現 57
第五節、補充精胺酸對於心臟蛋白表現的影響 61
一、觀察衰竭運動後左心室中Bcl-2蛋白的表現 61
二、觀察衰竭運動後左心室中connexin-43蛋白的表現 61
三、觀察衰竭運動後左心室中超過氧歧化酶蛋白的表現 61
第四節、以切片染色分析補充精胺酸對於心臟的影響 66
一、以Masson’s trichrome stain分析補充精胺酸對於心臟的影響 66
二、以DNA 片段分析補充精胺酸對於心臟的影響 66
三、以TUNEL stain分析補充精胺酸對於心臟的影響 66
第陸章 討論 71
第柒章 結論 78
第捌章 參考文獻 79
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