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研究生:周琬瑜
研究生(外文):Wan Yu Chou
論文名稱:低氧運動訓練對於嗜中性白血球殺菌效益之影響
論文名稱(外文):Effect of Hypoxic Exercise Training on Bactericidal Capacity of Neutrophil
指導教授:王鐘賢王鐘賢引用關係
指導教授(外文):J. S. Wang
學位類別:碩士
校院名稱:長庚大學
系所名稱:復健科學研究所
學門:醫藥衛生學門
學類:復健醫學學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
論文頁數:155
中文關鍵詞:低氧運動訓練多形核性白血球補體活化吞噬作用呼吸氧爆作用凋亡
外文關鍵詞:hypoxiaexercisetrainingpolymorphonuclear leukocytecomplement activationphagocytosisrespiratory burstapoptosis
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背景與目的:血液當中嗜中性白血球在發炎過程中佔有很重要的
角色; 主要形成體內第一道防線來抵抗外來侵入的微生物。雖然單次
急性低氧運動已被證實,能夠引起嗜中性白血球調節發炎過程;然而
長期低氧運動訓練對於嗜中性白血球的殺菌能力,目前仍不清楚。本
篇研究主要目的研究長期低氧運動訓練,對於嗜中性白血球的殺菌能
力之影響及相關分子的機制。
方法:60 位健康靜態生活的年輕男性,進行低氧合併運動的固
定式腳踏車訓練,分別執行五種訓練模式:兩種運動訓練(在百分之
十五氧氣濃度之下,分別合併百分之六十最大攝氧量及百分之六十的
心跳儲備量運動強度)、單純低氧訓練(百分之十五氧氣濃度)、單純
運動訓練(百分之六十最大攝氧量),以及控制組。訓練時間每天運
動二十分鐘,每週運動五天,共進行四週。於介入前和介入後立即與
兩小時後抽取前臂靜脈血液,並以流式細胞儀檢測嗜中性白血球的噬
菌作用、呼吸氧爆作用及移行作用、與其細胞膜之黏著分子(CD62L,
CD11b, CD11a)、補體接受器(CD11b, CD35 and CD88)、Fc III receptor
(CD16b)、清除受器(CD36)之表現,以及嗜中性白血球之凋亡相關現
象;包括細胞膜磷脂質絲胺酸外翻及凋亡蛋白3 的檢測。
結果:在訓練之前,劇烈運動能提升大腸桿菌刺激的嗜中性白血
iii
球吞噬作用、氧爆作用、移行作用;並且增加嗜中性白血球CD62L、
CD11b、CD11a、CD16b、CD93 及CD88 的表現。在凋亡表現方面,劇
烈運動會減少嗜中性白血球細胞膜磷脂質絲胺酸外翻及凋亡蛋白3
的表現。然而在經過四週的低氧運動訓練後,會減弱由劇烈運動引起
的嗜中性白血球殺菌能力與調理/補體接受器以及嗜中性白血球凋亡
的反應,低氧運動訓練有效提昇先天免疫殺菌能力的閾值。
結論:雖然劇烈運動可藉由增加調理/補體接受器的表現,提升嗜
中性白血球的功能,減少凋亡表現;但長期低氧運動訓練之後,會提
昇先天免疫殺菌能力的閾值,減少急性運動對於嗜中性白血球殺菌能
力的改變。
臨床意義:低氧運動訓練為有效訓練的訓練方式,藉由調節嗜中性
白血球吞噬相關的分子表現提昇嗜中性白血球殺菌作用之閾值。
Background and Purpose: Blood neutrophil play an important role in the
inflammation process and are the first-line defense to against invading
microorganisms. Although hypoxic exercise can enhance neutrophilmodulated
inflammation process at acute stage, the effects of long-term
hypoxic exercise training in bactericidal capacity of neutrophils remain
unclear. This study investigates the effect of long-term hypoxic exercise
training on bactericidal capacity of neutrophils and further explores the
underlying mechanisms.
Methods: Healthy sedentary young men were trained on a bicycle
ergometer, engaging five groups in a normobaric hypoxia chamber: two
hypoxic exercise training group (i.e., 15% O2 with 60%VO2max and
60%HRR), hypoxic training group (i.e., 15% O2), exercise training group
(60%VO2max), and control group. Duration of training is 20 minutes per
day, 5 days per week for 4 weeks. Phagocytosis, oxidative burst activity
and transmigration of neutrophil as well as expressions of adhesion
molecules (i.e., CD62L, CD11b and CD11a), complement receptors
(CD35 and CD88), FcγIIIB receptor (CD16b), and scavenger receptor
(CD36) on neutrophil were measured. Annexin-V binding to
phosphatidylserine (PS) and caspase 3 expression were used as markers
of neutrophil apoptosis. All results were measured by using flow
cytometery.
Results: Before training, strenuous exercise enhanced the extents of
E.coli-induced phagocytosis, oxidative burst activity and transmigration
of neutrophil, which was accompanied by the increases in expressions of
CD62L, CD11b, CD11a, CD16b, CD93 and CD88 on neutrophils.
Therefore, PS exposure and caspase 3 expressions decreased immediately
after strenuous exercise. However, 4 weeks of hypoxic exercise training
attenuated the enhancements of neutrophil bactericidal capacity,
opsonic/complement receptor and apoptosis expressions by acute
strenuous exercise. Hypoxic exercise training promoting the threshold of
neutrophil bactericidal capacity.
Conclusion: Although strenuous exercise can evoke the phagocytosis,
oxidative burst activity and transmigration of neutrophil by increasing the
expressions of opsonic/complement receptors and decreasing apoptosis
expression on neutrophil, long-term hypoxic exercise training increasing
the threshold of innate immune and suppresses the acute exercise effect
v
on the neutrophil bactericidal capacity.
Clinical relevance: Hypoxic exercise regimen is a good strategy for
promoting the threshold of bactericidal capacity which modulated by
different phagocytotic molecular expression.
論文目錄
誌謝 ... i
摘要 ... ii
Abstract ... iv
目錄 ... vi
圖表目錄 ... viii
第一章 緒論(Introduction)... - 1 -
第一節 研究背景及目的 ... - 1 -
第二節 研究假設 ... - 4 -
第二章 文獻回顧(Literature Review)... - 5 -
第一節 低氧運動訓練簡介 ... - 5 -
第二節 低氧環境下的生理適應反應 ...- 7 -
第三節 低氧與氧化壓力的相關性 ... - 10 -
第四節 免疫系統和發炎的相關性 ... - 12 -
第五節 補體系統在發炎反應中的角色 ...- 14 -
第六節 嗜中性白血球(Neutrophils)與發炎反應的關聯 ... - 19 -
第七節 低氧、運動和先天免疫參與發炎作用的相關反應 ... - 27 -
第三章 實驗設計(Experimental Design) ... - 34 -
第一節 實驗材料 ... - 34 -
第二節 實驗方法 ...- 39 -
受試者 ... - 39 -
實驗流程 ... - 39 -
多形核性白血球萃取 ... - 40 -
細胞膜上補體受器與黏著分子的測定 ...- 41 -
嗜中性白血球的吞噬作用測定 ... - 42 -
嗜中性白血球的氧爆作用測定 ...- 43 -
嗜中性白血球的移行作用測定 ... - 44 -
偵測細胞膜phosphatidylserine residues (PS) 外翻試驗- 44 -
活化凋亡蛋白3 試驗 ... - 45 -
統計分析 ... - 47 -
第四章 結果(Result) ...- 48 -
第一節 受詴者基本資料、身體狀況和活動情形調查 ...- 48 -
第二節 不同低氧運動訓練對於運動表現之影響 ...- 49 -
第三節 不同低氧運動訓練對於血球數目之影響 ... - 50 -
第四節 不同低氧運動訓練對於白血球黏著分子之影響 ... - 51 -
第五節 不同低氧運動訓練對於白血球的抗體受器、補體受器及清除受
器之影響 ... - 57 -
第六節 不同低氧運動訓練對於嗜中性白血球細胞的行為影響 .. - 66 -
第七節 不同低氧運動訓練對於嗜中性白血球細胞的凋亡影響 .. - 70 -
第五章 討論 (Discussion) ...- 75 -
低氧運動訓練對於人體測量數據的影響 ... - 75 -
低氧運動訓練對於最大體能的影響 ... - 75 -
低氧運動訓練對於先天免疫反應的影響 ... - 76 -
低氧運動訓練對於嗜中性白血球數目之影響 ...- 77 -
低氧運動訓練對於嗜中性白血球黏著之影響 ...- 78 -
低氧運動訓練對於嗜中性白血球抗原受器、補體受器及清除受器表現之
影響 ... - 80 -
低氧運動訓練對於嗜中性功能性表現之影響 .... - 82 -
低氧運動訓練對於嗜中性白血球凋亡表現之影響 ... - 88 -
第六章 結論 (Conclusion) ... - 90 -
參考文獻(References) ... - 129 -
附錄一 ... - 130 -
附錄二 ... - 130 -
圖表目次
表一 受試者基本資料 ... - 91 -
表二 五組訓練對於劇烈運動刺激下體能表現之影響 ... - 92 -
表三 五組訓練前後對於劇烈運動刺激白血球數目的改變 ... - 93 -
表四 五組訓練前後對於劇烈運動刺激及合併大腸桿菌刺激下白血球CD62L 的螢光表現 ...- 94 -
表五 五組訓練前後對於劇烈運動刺激及合併大腸桿菌刺激下白血球CD11b 的螢光表現...- 95 -
表六 五組訓練前後對於劇烈運動刺激及合併大腸桿菌刺激下白血球CD11a 的螢光表現...- 96 -
表七 五組訓練前後對於劇烈運動刺激及合併大腸桿菌刺激下白血球CD16b 的螢光表現...- 97 -
表八 五組訓練前後對於劇烈運動刺激及合併大腸桿菌刺激下白血球CD93 的螢光表現... - 98 -
表九 五組訓練前後對於劇烈運動刺激及合併大腸桿菌刺激下白血球CD35 的螢光表現... - 99 -
表十 五組訓練前後對於劇烈運動刺激及合併大腸桿菌刺激下白血球CD88 的螢光表現...- 100 -
表十一 五組訓練前後對於劇烈運動刺激及合併大腸桿菌刺激下白血球CD36 的螢光表現 ... - 101 -
表十二 五組訓練前後對於劇烈運動刺激白血球移行作用的表現 ... - 102 -
表十三 五組訓練前後對於劇烈運動刺激白血球吞噬作用的表現 ... - 103 -
表十四 五組訓練前後對於劇烈運動刺激及合併大腸桿菌刺激下白血球氧爆作用的表現... - 104 -
表十五 四組訓練前後劇烈運動刺激對於嗜中性白血球功能及分子表現之影響
...105 -
表十六 四組訓練後,對於嗜中性白血球功能及分子表現之影響 ... - 106 -
表十七 五組訓練前後對於劇烈運動刺激及合併大腸桿菌刺激下白血球磷脂質絲胺酸外翻的表現 ... - 107 -
表十八 五組訓練前後對於劇烈運動刺激及合併大腸桿菌刺激下白血球凋亡蛋白3 的表現 ...- 108 -
表十九 四組訓練前後劇烈運動刺激對於凋亡表現的改變 ... - 109 -
表二十 四組訓練後,對於凋亡表現的改變... - 110 -
圖一 發炎反應的步驟 ... - 111 -
圖二 探討發炎反應的分子機制 ... - 112 -
圖三 補體在發炎中反應中的角色 ... - 113 -
圖四 補體主要的四大功能 ... - 114 -
圖五 補體活化途徑 ... - 115 -
圖六 補體的古典途徑 ... - 116 -
圖七 補體的替代途徑 ... - 117 -
圖八 補體的膜攻擊複合體 ...- 118 -
圖九 補體的調理作用 ... - 119 -
圖十 嗜中性白血球 ... - 120 -
圖十一 嗜中性白血球的滾動、活化及黏著作用 ... - 121 -
圖十二 嗜中性白血球的移行作用 ... - 122 -
圖十三 嗜中性白血球的吞噬作用 ...- 123 -
圖十四 嗜中性白血球的氧爆作用 ...- 124 -
圖十五 實驗流程 ... - 125 -
圖十六 白血球的移行作用 ...- 126 -
圖十七 白血球的吞噬作用 ... - 127 -
圖十八 白血球的氧爆作用 ... - 128 -
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