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研究生:徐至清
研究生(外文):Jee-Ching Hsu
論文名稱:睡眠剝奪對於後繼的腦缺血具有制約效用:鼠海馬結構的神經傷害及膠細胞反應之研究
論文名稱(外文):Sleep deprivation elicits preconditioning effects on the subsequent cerebral ischemia: neuronal damage and glial reaction in the rat hippocampal formation
指導教授:李英雄藍琴臺藍琴臺引用關係
指導教授(外文):Ying-Shiung LeeChyn-Tair Lan
學位類別:博士
校院名稱:長庚大學
系所名稱:臨床醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:英文
論文頁數:110
中文關鍵詞:一氧化氮記憶海馬結構睡眠剝奪暫時性腦缺血微小膠細胞星狀膠細電子顯微鏡
外文關鍵詞:Nitric oxideMemoryHippocampal formationSleep deprivationTransient global cerebral ischemiaMicrogliaAstrogliaElectron microscopy
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本研究顯示在五天的睡眠剝奪(sleep deprivation)後,鼠海馬結構中一氧化氮(nitric oxide)的兩種專一性合成酶NADPH-d與NOS的染色標誌受到壓制,而微小膠細胞(microglia)與星狀膠細胞(astrocyte)的反應被激發。結論為長期睡眠剝奪所導致的記憶減退,其原因可能與一氧化氮的合成降低有關;而微小膠細胞與星狀膠細胞的激發,反映出長期睡眠剝奪後可能造成海馬結構的神經傷害。進一步用甲酚基紫(cresyl violet)染色及膠細胞表面抗原之免疫組織染色,探討睡眠剝奪對續發的腦缺血是否對鼠海馬結構產生影響。實驗結果顯示,暫時性腦缺血(transient global cerebral ischemia)恢復七天後引發的神經傷害與膠細胞反應,被五天的睡眠剝奪前處理所降低。最後用電子顯微鏡技術觀察未經與經過睡眠剝奪前處理的腦缺血鼠,發現引發的海馬結構的劇烈性超微構造破壞,也被睡眠剝奪前處理緩解了。據此,吾等推測睡眠剝奪對繼發性的致命性腦缺血可能具有前行制約(preconditioning)的效應。因此,睡眠剝奪或許可視為腦缺血傷害的預防及改善策略。

This study revealed that after sleep deprivation for 5 days, staining of two specific enzymes for nitric oxide synthesis, i.e., NADPH-d and NOS, were suppressed in rat hippocampal formation while microglia and astrocyes were activated. The inhibition of NADPH-d and NOS reactivities may account for the memory decline after long-term sleep deprivation. The concomitant induced reactions of microglia and astrocytes suggest the involvement of these cells in the deleterious effect in prolonged sleep deprivation. Furthermore, by using cresyl violet staining and immunohistochemistry detectiing glial cellular surface antigens, the influence of sleep deprivation on the subsequent global cerebral ischemia in rat hippocampal formation were studied. The results revealed that the ischemia-induced neuronal damages and glial reactions were ameliorated by pretreatment with sleep deprivation. Finally, the electron microscopic observations corroborated the above-mentioned finding in that sleep deprivation could greatly protect the hippocampal neurons from apoptosis induced by cerebral ischemia. It is postulated that sleep deprivation may have a preconditioning influence on subsequent lethal cerebral ischemia. Hence, sleep deprivation may be considered as a therapeutic strategy in brain ischemic damage.

指導教授推薦書
口試委員會審定書
授權書 iii
誌謝 iv
目錄 vi
中文總結 xi
英文總結 xii
第一部份 1
中文摘要 2
英文摘要 4
第壹章 導論 5
第貳章 材料與方法 8
第參章 結果 13
第肆章 討論 15
參考文獻 21
圖1. 在海馬結構中選取的細胞計數單位面積 29
圖2A-C. 正常鼠海馬結構NADPH-d反應陽性之神經元 30
圖3A-C. 在5天的睡眠剝奪後NADPH-d染色反應 31
圖4A-C. 正常鼠受OX-42標誌的微小膠細胞 32
圖5A-C. 經5天睡眠剝奪後被激發的微小膠細胞 33
圖6A-C. 一個正常鼠海馬結構中的星狀膠細胞 34
圖7A-C. 在睡眠剝奪鼠的海馬回激發之星狀膠細胞反應 35
圖8. 正常鼠及睡眠剝奪鼠海馬組織的西方墨點定量 36
圖9. 正常鼠及睡眠剝奪鼠的nNOS、OX-42與GFAP
西方墨點染色 37
表一、正常鼠及睡眠剝奪鼠每單位面積中NADPH-d、
OX-42及GFAP染色細胞之平均值與標準差 38
表二、nNOS、OX-42與GFAP免疫活性的平均密度 39
第二部份 40
中文摘要 41
英文摘要 42
第壹章 導論 43
第貳章 材料與方法 46
第參章 結果 50
第肆章 討論 54
參考文獻 62
圖1. 對照組腦缺血組及睡眠剝奪加腦缺血組中海馬結
構之甲酚基紫染色 71
圖2. 未經或經過睡眠剝奪前處理的腦缺血鼠受OX-42
標誌之微小膠細胞 72
圖3. 未經或經過睡眠剝奪前處理的腦缺血鼠的微小膠
細胞之OX-42免疫反應 73
圖4. 未經或經過睡眠剝奪前處理之腦缺血鼠受OX-18
標誌的微小膠細胞 74
圖5. 未經或經過睡眠剝奪前處理之腦缺血鼠受OX-6
標誌的微小膠細胞 75
圖6. 未經或經過睡眠剝奪前處理之腦缺血鼠受ED1
標誌的巨噬細胞/單核球系列細胞 76
圖7. 未經或經過睡眠剝奪前處理之腦缺血鼠受GFAP
染色之星狀膠細胞 77
圖8. 比較腦缺血組與睡眠剝奪加腦缺血組之OX-42、
OX-18、OX-6、ED1與GFAP免疫染色的平均
視覺密度 78
表一、對照組腦缺血組與睡眠剝奪加腦缺血組的神經細
胞損傷平均分數 79
表二、腦缺血組與睡眠剝奪加腦缺血組鼠每單位面積的
OX-42、OX-18、OX-6、ED1與GFAP受染細胞
數目之平均值與標準差 80
第三部份 81
中文摘要 82
英文摘要 83
第壹章 導論 84
第貳章 材料與方法 86
第參章 結果 88
第肆章 討論 90
參考文獻 95
圖1. 睡眠剝奪五天後鼠海馬回神經細胞的超微結構 100
圖2. 睡眠剝奪五天後之鼠海馬回神經氈 101
圖3. 睡眠剝奪五天後之鼠海馬回神經細胞 101
圖4. 腦缺血七天後鼠海馬回出現劇烈破壞性的形態改變 102
圖5. 腦缺血七天後之鼠海馬回中微小膠細胞伸出突起
包圍退化性神經細胞 103
圖6. 腦缺血七天後之鼠海馬回中微小膠細胞伸出突起
包圍退化性神經細胞 104
圖7. 腦缺血七天後之鼠海馬回中的神經氈 104
圖8. 腦缺血七天後之鼠海馬回的CA1區 105
圖9. 腦缺血七天後之鼠海馬回的神經末梢 105
圖10、11. 腦缺血七天後之鼠海馬回的神經末梢 106
圖12. 腦缺血七天後鼠海馬回微小膠細胞吞噬細胞碎屑 107
圖13. 睡眠剝奪加腦缺血之鼠海馬回退化性變化 107
圖14. 睡眠剝奪加腦缺血鼠之海馬回星狀膠細胞的突起
增生現象 108
圖15. 睡眠剝奪加腦缺血之鼠海馬回神經細胞 108
圖16. 睡眠剝奪加腦缺血鼠之海馬回偶見之微小膠細胞
吞噬現象 109
圖17. 睡眠剝奪加腦缺血鼠之海馬回偶見之微小膠細胞
吞噬現象 110

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第二部份
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