臺灣博碩士論文加值系統

背景與目的:源自骨髓的循環前驅細胞具有修復血管內皮損傷與血管新生的能力。雖然許多研究已證實，運動可提升週邊血液循環中循環前驅細胞的數量，但不同的運動模式對循環前驅細胞的影響仍未知。高強度有氧間隔式運動訓練已經被證明比等熱量的中低強度運動訓練對心血管功能和有氧代謝能力有更深的影響。本研究設計是想了解高強度有氧間隔式運動訓練如何影響循環前驅細胞，並與中強度持續性運動訓練比較對於缺血性低氧以及缺氧性低氧的適應能力，在細胞數量與功能上的變化。方法:收取32位無規律運動(一年內每週運動少於一次、每次運動少於20分鐘)的大學男性，隨機分配到1)有氧間隔式運動組：以最大攝氧量的40% 3分鐘和最大攝氧量的80% 3分鐘交替為一回合訓練，共進行5個回合。每天利用直立式腳踏車進行訓練30分鐘，一周5天，為期5周；2)中度持續性運動組：以最大攝氧量的60%持續訓練30分鐘。每天利用直立式腳踏車進行訓練30分鐘，一周5天，為期5周。運動訓練前後分別進行一次急性低氧運動測試(12%氧氣濃度，模擬海拔4500m之高山，以最大攝氧量60%做為運動強度，進行30分鐘腳踏車運動。)在急性低氧運動測試前後抽取週邊靜脈血液，使用流式細胞儀分析CD34+幹細胞、CD34+/KDR+循環血管新生細胞、CD34+/KDR+/CD117+造血幹細胞、CD34+/KDR+/ CD133+內皮前驅細胞、CD34+/KDR+/CD31+循環內皮細胞以及凋亡的循環血管新生細胞數量。再藉細胞培養方式，以顯微鏡及流式細胞儀觀察循環前驅細胞增生的能力。結果：無論是有氧間隔式運動或中度持續性運動訓練皆可提升心肺適能與體能表現。血液分析方面得知：首次急性低氧測試可提升血液中幹細胞、循環血管新生細胞、內皮前驅細胞數量。經五週訓練後，有氧間隔式運動訓練以幹細胞、循環血管新生細胞、內皮前驅細胞、循環內皮細胞增加為主。而中度持續性運動訓練則增加較多循環血管新生細胞。有氧間隔式運動訓練後，對於二次急性低氧測試，循環血管新生細胞抗凋亡的能力提升。結論：有氧間隔式運動訓練能加速幹細胞分化成較具血管新生功能的內皮前驅細胞並進入周邊血液循環中，並在經過低氧刺激後減少其凋亡的數量。臨床應用與未來研究方向：本實驗首次釐清有氧間隔式運動和中度持續性運動訓練對於循環前驅細胞重新分佈的影響。有關循環前驅細胞藉由物理方式刺激釋出後，在人體中分化、增生以及促進血管新生的能力，未來還需要更進一步的研究證實。希望運動治療能對內皮功能障礙的病患提供安全有效的治療策略。

Background and Purpose: Bone marrow-derived, circulating progenitor cells (CPCs) is contributing to the maintenance of endothelial function by mechanisms that endothelial repair to postnatal angiogenesis and vasculogenesis. Although exercise has been suggested that can upregulate CPCs number in blood, the effect of different mode of exercise on CPCs subsets remains unclear. High intensity aerobic interval training (AIT) has been shown to have a more profound influence on cardiovascular function and aerobic capacity than isocaloric moderate intensity continue training (MCT). The aim of this study was to investigate how AIT influences the mobilization of CPCs subsets into peripheral blood compartment. A second objective was to compare MCT with AIT that the adaptation of ischemic and hypoxic hypoxia changing in cell number and function. Methods: Thirty-two sedentary healthy men were participating in the study, which were randomly distributed to AIT exercise group (reciprocal 40%, 80%VO2max on a bicycle ergometer for 30min per day, 5 days per week, total 25 times) or MCT exercise group (60%VO2max). Acute severity hypoxia exercise testing was performed at pre and post chronic exercise training (60% VO2 max exercise intensity for 40 min under 12%O2 in air). At rest and immediately after severity hypoxia exercise, total of stem cells (SCs, CD34+ cells), circulating angiogenesis cells (CACs, CD34+/KDR+ cells), hematopoietic stem cells (HSCs, CD34+/KDR+/CD117+ cells), endothelial progenitor cells (EPCs, CD34+/KDR+/CD133+ cells), circulation endothelial cells (CECs, CD34+/KDR+/CD31+ cells), and the apoptosic cells number of CACs (CD34+/KDR+/phosphotidylserine exposed cells) were measured by three-color flow cytometry. Subsequently, we measured the ability of CPCs and subtypes cell proliferation by cell culture. Result：Exercise training improved cardiopulmonary fitness in AIT and MCT group. In blood analysis, acute hypoxia exercise testing could improve CACs and EPCs in first time. After five weeks training, AIT group increased the number of SCs, CACs and EPCs in peripheral blood compartment and MCT group significantly increased the number of CACs in the blood. Conclusion：Aerobic interval training markedly induced SCs differentiation to the functional CPCs into peripheral blood. Clinic Application and Future Research ：This study clarify the relationships between AIT and MCT modulated redistribution of CPCs. In future, require more research relevant to CPCs differentiation and proliferation capacity and improve angiogenesis ability in vivo by physical therapy method. We expect to provide a safe and effective strategy of exercise therapy for patients with endothelial dysfunction.

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國家圖書館 博碩士論文電子檔案上網授權書 ... iii
長庚大學博碩士論文著作授權書... iv
致謝 ... v
摘要 ... vii
Abstract ... ix
目錄 ... xi
第一章 緒論 ... -1-
一、研究背景 ... -1-
1.1 心血管疾病 ... -1-
1.2 血管內皮前驅細胞 ... -1-
1.3 幹細胞療法 ... -2-
1.4 非侵入式治療 ... -3-
二、研究動機與目的 ... -4-
第二章 文獻回顧 ... -6-
一、心血管患者的新希望 ... -6-
二、循環前驅細胞來自骨髓幹細胞 ... -6-
三、血管內皮生長因子接受器-2 為與血管新生有關之細胞標記 ... -6-
四、造血幹細胞的判定須靠多重細胞面標記輔以細胞培養觀察 ... -7-
五、內皮前驅細胞、循環中的內皮細胞 ... -7-
六、五種與內皮前驅細胞有關的重要激素 ... -9-
6.1 低氧誘發因子(HIF-1α) ... -9-
6.2 血管內皮生長因子(VEGF) ... -9-
6.3 基質細胞衍生因子(SDF-1) ... -10-
6.4 基質金屬蛋白酶(MMP-9) ... -10-
6.5 一氧化氮(NO) ... -10-
七、細胞培養內皮細胞 ... -11-
八、影響內皮前驅細胞數量與功能的因素 ... -11-
九、低氧對生理的影響 ... -12-
十、運動對生理的影響 ... -15-
十一、運動時局部肌肉組織氧氣力利用增加，使血液再分佈 ... -15-
十二、運動時心血管的調節 ... -15-
十三、運動時呼吸系統的調節 ... -16-
十四、運動對於循環前驅細胞的效果 ... -16-
十五、有氧間隔式運動訓練(Aerobic interval training) ... -18-
第三章 材料與方法 ... -19-
3.1 受試者 ... -19-
3.2 運動測試 ... -19-
3.2.1 最大運動測試 ... -19-
3.2.2 急性低氧運動測試 ... -21-
3.2.3 停止運動條件 ... -22-
3.3 肢段缺血再灌流測試 ... -22-
3.4 血液實驗 ... -23-
3.4.1 淋巴球萃取 ... -23-
3.4.2 CD34+幹細胞定量 ... -24-
3.4.3 CD34+/KDR+循環血管新生細胞定量 ... -24-
3.4.4 CD34+/KDR+/CD117+造血幹細胞定量 ... -25-
3.4.5 CD34+/KDR+/CD133+內皮前驅細胞定量 ... -26-
3.4.6 CD34+/KDR+/CD31+循環內皮細胞定量 ... -27-
3.4.7 循環血管新生細胞凋亡定量(CD34+/KDR+/PS+) ... -27-
3.5 體外細胞培養實驗 ... -28-
3.5.1 淋巴球萃取 ... -28-
3.5.2 細胞培養第 0 天 ... -29-
3.5.3 細胞培養第 3 天 ... -30-
3.5.4 細胞培養第 5 天 ... -30-
3.6 實驗設計 ... -35-
3.7 資料處理與分析 ... -37-
第四章 實驗結果 ... -38-
4.1 經五週有氧間隔式運動訓練或中度持續性運動訓練後，肺部通氣量與換氣效益的變化 ... -38-
4.2 有氧間隔式運動訓練和中度持續性運動訓練對心臟的影響 ... -39-
4.3 有氧間隔式運動訓練或中度持續性運動訓練對大腦血流的影響 ... -40-
4.4 有氧間隔式運動訓練可提升骨骼肌氧氣利用的效益 ... -40-
4.5 有氧間隔式運動訓練和中度持續性運動訓練後，整個體能表現與心肺適能的變化 ... -40-
4.6 有氧間隔式運動訓練和中度持續性運動訓練對於上下肢動脈血流的影響 ... -41-
4.7 有氧間隔式運動訓練可刺激 CD34+骨髓幹細胞釋出 ... -42-
4.8 有氧間隔式運動訓練和中度持續性運動訓練可提升 CD34+/KDR+循環血管新生細胞數量 ... -42-
4.9 急性低氧測試可使 CD34+/KDR+/CD117+造血幹細胞數量增加 ... -43-
4.10 有氧間隔式運動訓練可提升 CD34+/KDR+/CD133+內皮前驅細胞數量 ... -43-
4.11 有氧間隔式運動訓練可提升血液 CD34+/KDR+/CD31+循環內皮細胞數量 ... -43-
4.12 有氧間隔式運動訓練可提升與血管新生相關細胞抗凋亡的能力 ... -44-
4.13 有氧間隔式運動訓練和中度持續性運動訓練對於內皮前驅細胞培養後增生速率的影響 ... -44-
第五章 討論 ... -45-
5.1 運動對心血管疾病的重要性 ... -45-
5.2 有氧間隔式與中度持續性運動訓練對增進心肺適能的影響 ... -46-
5.3 運動可使循環前驅細胞更具方向性 ... -46-
5.4 觀察有氧間隔式和中度持續性運動訓練對於循環前驅細胞的變化 ... -46-
第六章 結論 ... -49-
圖表 ... -50-
圖一 有氧間隔式運動訓練組和中度持續性運動訓練組訓練模式 ... -50-
圖二 實驗流程圖 ... -51-
圖三 99 年台灣十大死因人數圖 ... -52-
圖四 有氧間隔式運動訓練組和中度持續性運動訓練組訓練前後循環系統變化 ... -53-
圖五 有氧間隔式運動訓練組和中度持續性運動訓練組訓練前後大腦血流氧氣利用情形 ... -54-
圖六 有氧間隔式運動訓練組和中度持續性運動訓練組訓練前後肌肉血流氧氣利用情形 ... -55-
圖七 有氧間隔式運動訓練組和中度持續性運動訓練組運動訓練前後在基礎動脈血流的變化 ... -56-
圖八 有氧間隔式運動訓練組和中度持續性運動訓練組運動訓練前後在缺血再灌流動脈血流的變化 ... -57-
圖九 有氧間隔式運動訓練組和中度持續性運動訓練組運動訓練前後肌肉攝氧量的變化 ... -58-
圖十 有氧間隔式運動訓練組和中度持續性運動訓練組運動訓練前後動脈再充氧的變化 ... -59-
圖十一 有氧間隔式運動訓練組和中度持續性運動訓練組運動訓練前後 CD34+幹細胞的數量 ... -60-
圖十二 有氧間隔式運動訓練組和中度持續性運動訓練組運動訓練前後 CD34+/KDR+循環血管新生細胞的量
... -61-
圖十三 有氧間隔式運動訓練組和中度持續性運動訓練組運動訓練前後 CD34+/KDR+/CD117+造血幹細胞的數量
... -62-
圖十四 有氧間隔式運動訓練組和中度持續性運動訓練組運動訓練前後 CD34+/KDR+/CD133+內皮前驅細胞的數量
... -63-
圖十五 有氧間隔式運動訓練組和中度持續性運動訓練組運動訓練前後 CD34+/KDR+/CD31+循環內皮細胞的數量
... -64-
圖十六 有氧間隔式運動訓練組和中度持續性運動訓練組運動訓練前後與血管新生相關細胞凋亡(CD34+/KDR+/PS+)
的數量 ... -65-
表一 從骨髓幹細胞到成熟內皮細胞表面抗原標記演化表 ... -66-
表二 受試者基本資料 ... -67-
表三 有氧間隔式運動訓練組和中度持續性運動訓練組運動訓練前後心臟功能的變化 ... -68-
參考文獻 ... -69-
附錄 ... -74-
附錄一 受試者參與實驗同意書 ... -74-
附錄二 身體狀況與活動情形調查表 ... -78-

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