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題名：長期分解性壓力對骨骼肌AMPK訊息系統之影響
作者：陳宗與
作者(外文)：Chung-Yu Chen
校院名稱：國立體育大學
系所名稱：體育研究所
指導教授：蔡櫻蘭
郭家驊
學位類別：博士
出版日期：2010
主題關鍵詞：糖尿病；醣類代謝；運動；肥胖；葡萄糖轉運體；diabetes；carbohydrate metabolism；exercise；obesity；glucose transporter
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低氧刺激與運動訓練兩種模式已被證實能改變骨骼肌系統代謝功能，目前了解，骨骼肌醣類代謝狀態會受到AMPK與CAMK兩訊息路徑系統調節。因此，本研究第一個實驗為探討不同輕微低氧曝露時程對AMPK訊息路徑與胰島素敏感度之影響；第二個實驗則是比較急性運動與運動訓練對肥胖Zucker鼠葡萄糖耐受度之影響，同時探討AMPK訊息路徑與CAMK II蛋白所扮演之角色。方法：本研究第一個實驗將56隻SD (Spargue Dawley)鼠依體重配對方式分成每日低氧8小時與1小時兩大組，各大組則再細分為控制組、單次低氧組與八週低氧組。第二個實驗將24隻Zucker胖鼠與24隻Zucker瘦鼠依體重配對方式分成胖與瘦兩大組，每大組又細分為控制組、單次運動1小時組及六週長期運動訓練組。結果：在實驗一部份，我們發現每天8小時且連續八週間歇性低氧能明顯減低副睪脂肪量、改善葡萄糖耐受度、增加胰島素敏感度與增加微血管密度；連續八週每天1小時低氧後，僅在胰島素敏感度與微血管密度與控制組比較後有差異。在基因表現部份，單次低氧8小時後明顯增加PGC-1α、VEGF及GLUT4基因表現，在AMPK-PGC-1α-GLUT4及VEGF蛋白表現部份，低氧8小時和1小時兩大組與控制組比較後無明顯差異。在實驗二部份，我們發現長期運動訓練明顯改善Zucker鼠葡萄糖耐受度與增加磷檬酸合成酶的活性，但在GLUT1、GLUT4、PGC-1α基因表現上，單次1小時運動後對Zucker鼠無明顯影響。胖與瘦Zucker鼠在基礎狀態下之AMPK α-thr 172與CAMK II-thr 286磷酸化和CAMK II蛋白表現即有明顯差異，但運動訓練後胖鼠能明顯趨於正常化表現狀態，在PGC-1α蛋白表現部份，胖與瘦Zucker鼠在基礎狀態下有明顯差異，瘦鼠在六週運動訓練後明顯提高PGC-1α蛋白表現。結論：本研究實驗一發現每天低氧8小時且連續八週後明顯改善葡萄糖耐受度與胰島素敏感度，但此正面效益在每天低氧1小時組較不明顯。連續八週每天低氧8小時所帶來改善胰島素敏感度與葡萄糖耐受度之效果，可能與降低脂肪量和增加微血管密度有關，似乎與GLUT4、VEGF、和PGC-1α蛋白表現沒有關連。實驗二發現在基礎狀態下，肥胖Zucker鼠與瘦Zucker鼠之CAMK II-thr 286磷酸化與CAMK II蛋白表現、AMPK α-thr 172磷酸化有明顯差異，但經由六週的游泳訓練後能明顯使此現象趨向於正常化(normalize)。運動訓練改善肥胖Zucker鼠葡萄糖耐受度的效果，部份可能的原因與增加粒線體密度有關，似乎與GLUT1、GLUT4、PGC-1α蛋白表現無直接關連。本研究的結果將有助於了解長期間歇性低氧與運動訓練對骨骼肌醣類代謝調節機轉之影響，此結果在未來可做為治療或改善葡萄糖耐受度與胰島素敏感度的應用策略。
Hypoxia and exercise training are the well known factors, which can cause functional changes in skeletal muscle. Glucose metabolism in skeletal muscle is mediated by AMPK and CAMK II signaling pathway, which is crucial in normal and obese subjects. The first study was purposed to investigate the different duration of mild hypoxia exposure on AMPK signaling and insulin sensitivity. The second study was aimed to compare the acute exercise and exercise training effect on glucose tolerance in obese Zucker rat. This study also demonstrate the role of AMPK signaling and CAMK II protein expression in Zucker rats. Methods: In the first study, Spargue Dawley (n=56) rats were divided into two groups; 8 hr/day mild hypoxia exposure group and 1 hr/day mild hypoxia exposure group. Each group was followed 3 treatments: control, acute mild hypoxia and 8-week mild hypoxia. In the second study, obese Zucker rats (n=24) and lean littermates (n=24) were assigned into following 3 treatments; control, acute 1-hr exercise and six-week chronic exercise training. Results: In first part of the study, significantly decreased epididymal fat mass, improved glucose tolerance, enhanced insulin sensitivity, and increased capillary density was observed with 8 hr/day with 8-week mild hypoxia treatment. Only in insulin sensitivity and capillary density were significantly increased in 1 hr/day with 8-week mild hypoxia treated group. Furthermore, acute 8-hour mild hypoxia exposure significantly elevated PGC-1α, VEGF and GLUT4 mRNA levels. However, the protein expression of AMPK, phospho-AMPK, PGC-1α, GLUT4 and VEGF in both red and white quadriceps muscles were not significantly different among groups under acute and chronic mild hypoxia conditions. In second study, exercise training significantly improved glucose tolerance and increased citrate synthase activity in both obese and lean Zucker rats. However, no significant difference in GLUT1, GLUT4, and PGC-1α mRNA levels were observed in red gastrocnemius muscle between the control and acute exercise groups. Interestingly, the phosphorylation of AMPK-thr 172, CAMK II-thr 286, and PGC-1α protein level in obese Zucker rat were significantly higher than lean littermates under basal state, but this phenomenon could be reversed after six-week exercise training. In addition, the PGC-1α level was significantly increased in lean Zucker rats following six-week exercise training. Conclusions: The first study demonstrated significant improvements in glucose tolerance and insulin sensitivity by 8 hr/day with 8-week mild intermittent hypoxia. However, this beneficial effect was not apparent in 1 hr/day with 8-week mild intermittent hypoxia group. This improvement in glucose tolerance and insulin sensitivity appears to be related to the elevated muscle capillary density and decreased fat accumulation; it seems not to be related with increased GLUT4, VEGF, and PGC-1α protein expression. The second study showed significantly higher CAMK II-thr 286 and AMPK-thr 172 phosphorylation in obese Zucker rat than in lean rats; however, six weeks exercise training could reverse this phenomenon. Improved glucose tolerance by exercise training in both obese and lean Zucker rats appear to be related to increased mitochondria density rather than elevated GLUT1, GLUT4, and PGC-1α protein expression. This study confirmed that mild intermittent hypoxia and exercise training would be affective to improve glucose tolerance and increase insulin sensitivity, which can adopt as therapeutic strategy for type 2 diabetic patients.
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