臺灣博碩士論文加值系統

　　本研究目的在探討蜂膠之咖啡酸苯乙酯對人類白血球在高溫環境下歷經24小時後耐受度之影響。受試者分別為參加專業自由車訓練達三年以上的優秀自由車選手（三名男性，平均年齡20.00±0.00歲，平均身高175.67±3.48公分，平均體重69.00±0.58公斤）與無規律運動習慣之對照組（三名男性，平均年齡20.67±0.33歲，平均身高176.00±1.73公分，平均體重64.67±3.33公斤）。所有受試者皆採集安靜時血液，分離單核白血球（mononclear cells）並加入不同濃度之蜂膠的有效成份－咖啡酸苯乙酯（caffeic acid phenethyl ester, CAPE）分別置於37℃及43℃（1小時）培養箱中歷經24小時後進行生長曲線、形態學、機能學分析。另外，以流式細胞儀測定細胞內glutathione及superoxide anion的含量。每位受試者必須接受原地型腳踏車運動測驗以測得最大攝氧量及採集安靜狀態之血液樣本進行生化分析。研究結果發現，在基礎生理、生化值中，運動員的最大攝氧量與安靜時肌酸激值較非運動員高、運動後血乳酸峰值表現則較非運動員低。高溫環境會使人體單核細胞於生長曲線變化中的存活數下降；於形態學中的壞死率上升；於機能學變化中的NBT還原試驗之陽性率上升；在氧化還原測定中superoxide含量增加。蜂膠之咖啡酸苯乙酯（CAPE）除不會改變gultathione含量外，能有效地保護人體單核細胞在高溫環境下所受的損傷，包括：生長曲線隨濃度增加其存活數上升；形態學中隨濃度增加其壞死率有下降之趨勢；降低細胞釋放出superoxide的能力。另外，無論有無高溫環境、蜂膠之咖啡酸苯乙酯處理，運動員的單核細胞存活數、壞死率與superoxide含量較非運動員低外，還具有放出大量superoxide的能力。綜合上述，本研究證實，保持長期運動訓練的優秀自由車男性選手之單核細胞對高溫環境的容忍度較一般健康人高外，蜂膠之咖啡酸苯乙酯可保護高溫環境對運動員與非運動員單核細胞的傷害。由此對一般大眾提出有效地於高溫環境下的對策外，也提供運動員於參加競賽或訓練時的參考，進而締造佳績。

The purpose of this research is to determine the effect of propolis on the tolerance of human leukocytes exposed to hyperthermia. The Subjects were separated into two groups. The first group was well-trained competitive cyclists (n=3, male, 20.00±0.00 years, 175.67±3.48 cm, 69.00±0.58 kg) who had trained more than 3 years; the second group was sedentary controls (n=3, male, 20.67±0.33 years, 176.00±1.73 cm, 64.67±3.33 kg) with no special training experience. We isolated peripheral blood mononuclear cells (MNC) from the subjects, and treated MNC with various doses of caffeic acid phenethyl ester (CAPE); MNC were separated into two subjects；the first was incubated at 37℃, the second was incubated at 43℃; after 24 hours, the proliferation, morphological changes and functional changes were assessed. We used flow cytometry to detect intracellular gultathione and superoxide anion content. Each subject underwent a stationary bicycle test for VO2 peak. Their blood was sampled for biochemical analysis after rest. The results show that the values of VO2 peak and resting creatine kinase (CK) in cyclists are higher than those of sedentary controls. However, the post-exercise peak value of blood latic (BLa) in cyclists is lower than controls. The hyperthermic effects on MNC of both cyclists and controls include as follows: decrease in viability, increase in necrosis, augmentation in superoxide production and increase in intracellular superoxide content, CAPE treatment can prevent the hyperthermia-induced changes above. Neither hyperthermia nor CAPE significantly changes intracellular glutathione content of MNC. Moreover, the viability, necrotic ratio and superoxide content of MNC in cyclists are lower than controls with or without hyperthermia/CAPE treatment. We condude that the MNC from cyclists is more tolerable to hyperthermic injury and CAPE which may protect MNC from both cyclists and sedentary controls against this injury.

目　　錄
致謝…………………………………………………………………I
中文摘要……………………………………………………………II
英文摘要……………………………………………………………III
目錄…………………………………………………………………IV
第一章　緒論………………………………………………………1
　一、前言…………………………………………………………1
　二、研究背景與動機……………………………………………2
　三、研究目的……………………………………………………4
　四、研究假設……………………………………………………6
　五、研究範圍與限制……………………………………………6
　六、名詞操作性定義……………………………………………7
第二章　文獻探討…………………………………………………9
　一、蜂膠與咖啡酸苯乙酯………………………………………9
　　（一）蜂膠的定義及來源……………………………………9
　　（二）蜂膠之基本物理性質…………………………………10
　　（三）蜂膠與咖啡酸苯乙酯的組成份………………………11
　　（四）蜂膠與咖啡酸苯乙酯的效用…………………………12
　　　　1.抗腫瘤、抗誘突變作用………………………………12
　　　　2.抗發炎作用……………………………………………12
　　　　3.抗菌作用………………………………………………13
　　　　4.抗病毒作用……………………………………………13
　　　　5.抗氧化作用……………………………………………13
　　　　6.免疫活化作用…………………………………………14
　　　　7.蜂膠與咖啡酸苯乙酯之其他活性……………………14
　　　　8.蜂膠與咖啡酸苯乙酯之生理療效……………………15
　　（五）小結……………………………………………………16
　二、運動與免疫的關係…………………………………………16
　三、運動與白血球的關係………………………………………20
　　（一）運動與淋巴球的關係…………………………………21
　　（二）運動與毒殺細胞的關係………………………………24
　　（三）運動對人體血液中白血球、淋巴球、單核球等數量之影響30
　　（四）小結……………………………………………………34
　四、運動、人體免疫系統與高溫環境的關係…………………36
　　（一）運動與高溫環境之探討………………………………37
　　（二）高溫環境與免疫功能之探討…………………………41
　　（三）小結……………………………………………………43
第三章　材料與方法………………………………………………45
　一、實驗材料……………………………………………………45
　　（一）儀器設備………………………………………………45
　　（二）實驗耗材用品…………………………………………47
　　（三）化學試劑………………………………………………48
　　（四）細胞製備………………………………………………48
　二、實驗方法……………………………………………………48
　　（一）受試者…………………………………………………48
　　（二）運動能力檢測項目流程………………………………49
　　（三）單核細胞之測定及分析………………………………50
　　（四）蜂膠中咖啡酸苯乙酯之製備…………………………54
　　（五）血液抽取程序…………………………………………54
　　（六）資料處理………………………………………………54
第四章　結果………………………………………………………55
　一、受試者基本資料……………………………………………55
　二、高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之生長曲線分析…………………………………………………………………55
　三、高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之形態學分析…………………………………………………………………56
　四、高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之機能學分析…………………………………………………………………57
　五、高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之氧化還原測定…………………………………………………………………57
　　（一）細胞內gultathione含量……………………………58
　　（二）細胞內superoxide含量……………………………58
第五章　討論………………………………………………………60
　一、實驗模式探討………………………………………………61
　　（一）高溫環境條件之探討…………………………………61
　　（二）單核細胞之探討………………………………………62
　　（三）蜂膠中咖啡酸苯乙酯角色之探討……………………64
　二、實驗結果探討………………………………………………66
　　（一）受試者基本資料………………………………………66
　　（二）高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之生長曲線的影響……………………………………………67
　　（三）高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之形態學的影響…………………………………………………68
　　（四）高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之機能學的影響…………………………………………………70
　　（五）高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之氧化還原的影響………………………………………………71
　　　　　1.細胞內gultathione含量之探討…………………71
　　　　　2.細胞內superoxide含量之探討……………………72
第六章　結論與建議………………………………………………75
　一、結論…………………………………………………………75
　二、建議…………………………………………………………76
表……………………………………………………………………77
圖……………………………………………………………………83
參考文獻……………………………………………………………97
附錄…………………………………………………………………114
表　　次
表一：受試者基本資料與基礎生理、生化值表…………………77
表二：高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之生長曲線分析……………………………………………………78
表三：高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之形態學分析………………………………………………………79
表四：高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之機能學分析………………………………………………………80
表五：高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之氧化還原測定－細胞內gultathione含量………………………81
表六：高溫環境與不同濃度咖啡酸苯乙酯對人類白血球之氧化還原測定－細胞內superoxide含量………………………82
Figure Content
Fig 1:Growth inhibitory……………………………………83
Fig 2:Morphology of lymphocytes…………………………84
Fig 3:Morphology of monocytes……………………………85
Fig 4:NBT reduction test…………………………………86
Fig 5:Intracellular GSH content of lymphocytes……87
Fig 6:Intracellular GSH content of monocytes………88
Fig 7:Intracellular superoxide content of lymphocytes…………89
Fig 8:Intracellular superoxide content of monocytes90
Fig 9:Morphology of mononuclear cells （MNC）obsered under light microscope (400X)－37℃……………………………91
Fig10:Morphology of mononuclear cells （MNC）obsered underlight microscope (400X)－43℃, 1hr…………………92
Fig11:Morphology of mononuclear cells （MNC）obsered under light microscope (400X)－CAPE than 43℃, 1hr…………93
Fig12:NBT reduction test of MNC obsered under light microscope (400X)－37℃………………………………………94
Fig13:NBT reduction test of MNC obsered under light microscope (400X)－43℃, 1hr…………………………………95
Fig14:NBT reduction test of MNC obsered under light microscope (400X)－CAPE than 43℃, 1hr……………………96
附　　錄
附錄一：受試者須知…………………………………………114
附錄二：受試者同意書………………………………………115
附錄三：健康情況調查表………………………………………116

一、中文部份
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吳粹文、張復興、方兵兵、李熠、丁烈佛、張國南（1997），真理蜂膠營養液抗疲勞作用實驗研究，中國養蜂，6，9-10。
吳梅桂（1997），檳榔嚼塊去除口臭及其氧化特性之探討，中山醫學院營養科學研究所，碩士論文。
李文源、陳莉萍（1992），蜂膠酊治療外耳道炎的臨床和實驗觀察，密蜂雜誌，2，3-4。
李浤岦（1997），熱休克前置處理對過敏性休克的心臟循環功能保護作用之探討，高雄醫學院醫學研究所碩士論文。
李樹榮、尹培輝、王豔芬（1997），蜂膠制劑對羊鼻繩幼蟲禽體殺滅試驗，養蜂科技，6，5-6。
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林銘隆（1998），男性肥胖者在不同強度下運動中心管及體溫調節反應之比較研究，國立體育學院運動科學研究所碩士論文。
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茅力、楊森、陳景衡、金念組、魏國勤（1998），蜂膠醇溶液營養成分分析，南京醫科大學學報，18 (6), 543-544。
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陳惠英（1996），茶葉萃取物之抗突變及抗氧化特性，國立中央大學食品科學研究所，博士論文。
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童麗霞（2000），蜂膠抗氧化性之研究，中山醫學院營養科學研究所，碩士論文。
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黃文誠（1997），蜂膠對小白鼠腹水癌的抗腫瘤作用，蜜蜂雜誌，1，7。
黃永任（2000），運動與乳癌的預防，國民體育季刊，29，(1)，68-74。
黃森芳（1998），運動對人體免疫功能之影響，教育部國民體育季刊，27 (2)，81-89。
黃森芳（民87），運動對人體免疫系統中自然殺手細胞功能之影響，教育部國民體育季刊，27 (1)，32-45。
黃森芳（民88），一次運動後血液中淋巴細胞稀少現象Cortisol可能扮演之角色，教育部國民體育季刊，28 (3)，86-94。
黃森芳（民88），一次運動對人體血液中白血球及淋巴球數量之影響，教育部國民體育季刊，28 (1)，62-79。
黃衛平、劉放（1998），蜂膠的研究進展，養蜂科技，5，6-11。
楊世達、豐東洋、黃廖植、林倩華、周啟雄、陳俊忠（1995），高溫環境下最大衰竭運動之溫度調節現象探討，體育學報，20，399-410。
楊朝環（1999），蜂產品民間應用法，蜜蜂雜誌月刊，6，14。
楊瑞成（1997），熱休克前置處理對過敏性休克的心臟循環功能保護作用之探討，高雄醫學院醫學研究所碩士論文。
解玉珍（1999），不同種類蜂膠之組成分分析及抗生特性研究，大業大學食品工程研究所，碩士論文。
廖大昆（1999），用蜂膠治療高脂血症，蜜蜂雜誌，3，7。
劉家全（1999），醣皮質類固醇可延長熱中風大白鼠之存活時間，國立陽明大學生理學研究所碩士論文。
劉淑珍（1991），蜂膠是萬金油還是一種法定藥品，蜜蜂雜誌，3，11。
劉增城（1997），蜜蜂如何採膠利用，蠶蜂業專訊，p.14-17。
德水勇治郎（1996），蜂膠可去除體內毒素，書春出版社，台北市。
駱尚驊（1999），蜂膠質量控制的幾個指標和檢測方法，中國養蜂，50 (3)，31。
顏貽本（1996），蜂膠對血小板凝聚的體外效應，蜜蜂，3，19。
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二、英文部份
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