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研究生:王品雯
研究生(外文):Pin-Wen Wang
論文名稱:以小鼠探討性別及停經因子影響間歇性缺氧引發認知功能損害之病理機轉
論文名稱(外文):The Pathological Mechanisms of Sex and Menopause Factors Affecting Intermittent Hypoxia-Induced Cognitive Impairments in Mice
指導教授:楊靜修楊靜修引用關係郭博昭郭博昭引用關係
指導教授(外文):Cheryl C.H. YangTerry B.J. Kuo
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:腦科學研究所
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:2020
畢業學年度:108
語文別:中文
論文頁數:58
中文關鍵詞:睡眠呼吸中止症間歇性缺氧更年期去卵巢母鼠認知功能損害自律神經功能睡眠結構腦波認知功能行為測試
外文關鍵詞:sleep apneaintermittent hypoxiamenopauseovariectomized micecognitive impairmentautonomic functionsleep architectureelectroencephalographycognitive task
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背景與目標:睡眠呼吸中止症 (sleep apnea, SA) 引發的間歇性缺氧 (intermittent hypoxia, IH) 和睡眠片斷化及高比例伴隨產生的自律神經失調、氧化壓力及免疫發炎等問題,均是造成失智症的風險因子之一。女性在更年期後歷經荷爾蒙極大的轉變,罹患阻塞型睡眠呼吸中止症 (obstructive sleep apnea, OSA) 比率會非常明顯的上升,然而性別及停經因子對於SA引發認知功能問題甚至失智症的影響程度及其中機轉仍不清楚。因此本研究擬以不同性別及有無去卵巢之母鼠探討此議題。假設:IH引發小鼠自律神經失調及睡眠片斷化,進而導致認知功能衰退,且去卵巢 (ovariectomized, OVX) 母鼠較無去卵巢 (sham-operated, Sham) 母鼠及公鼠更嚴重。材料與方法:本研究使用C57BL/6J的公鼠及Sham與OVX母鼠在16週時手術植入電極,一週後記錄24小時腦波、心電波及肌電波之無線生理訊號,並給予連續兩週,每天八小時,每小時30次的IH,同時每週記錄一次24小時生理訊號,進行後續分析自律神經及睡眠結構,接著進行新物體辨識與水迷宮認知測驗。結果:不論在公鼠、Sham或OVX母鼠中,IH相較空氣控制組在IH介入期間皆為睡眠總時間較短,清醒總時間較長,睡眠片斷次數較高,睡眠維持能力亦較差。此外,IH使公鼠在清醒和睡眠期的交感活性降低。在新物體辨識結果中,公鼠及Sham母鼠的工作記憶能力在經歷IH後皆顯著變差,而OVX母鼠的學習記憶本身就較差,沒有受到IH影響;在水迷宮結果中,IH的介入並沒有讓公鼠與Sham母鼠的空間學習記憶受到影響,但OVX母鼠在IH後認知功能與IH期間的睡眠品質呈顯著正相關,與睡眠中斷次數呈顯著負相關。結論:IH會使公母鼠的睡眠品質下降,公鼠與Sham母鼠的自律神經失調及認知功能表現下降,而OVX母鼠本身就具有較差的認知功能。
Background and Aims: Sleep apnea (SA) causes intermittent hypoxia (IH) and sleep fragmentation, which induces autonomic dysfunction, oxidant stress and inflammation. Those mentioned above are the risks of dementia. Women have increased incidence rate of obstructive sleep apnea (OSA) after menopause. However, it is still unclear whether and how gender and menopause factors will affect SA-induced cognitive impairments or even dementia. Therefore, the study aimed to explore differential mechanisms of SA-induced cognitive impairments between male, female sham-operated (Sham) and ovariectomized (OVX) mice. Hypothesis: IH causes autonomic dysregulation and sleep fragmentation, and further results in cognitive impairments in mice, which would be aggravated in OVX female mice. Materials and Methods: Experiment were carried out on 16-week-old mice. One week after the mice had electrodes implanted in the skulls, the 24-hour physiological signals were recorded wirelessly in freely moving mice. Male, female sham and OVX mice were exposed to 30 IH/hour for 8 hours during the light cycle daily for 2 weeks, and physiological signals were simultaneously measured. Moreover, Novel object recognition test and Morris water maze were performed to assess the cognitive function in mice. Results: In male, female sham and OVX mice, IH group spent less time and duration in quiet sleep (QS) and paradoxical sleep (PS), more time in active waking (AW), and had more interruptions of QS during IH intervention than did room air (RA) control group. Additionally, in male mice, sympathetic activity during AW and QS was lower in IH group than RA group. In the Novel object recognition test, IH group of male and sham female mice had worse working memory than did RA group. However, it had poor cognitive function in OVX female mice, so there was no difference between IH and RA. In the Morris water maze test, IH intervention had no impact on spatial learning memory in male and sham female, but correlation analysis had revealed that spatial memory was positive correlated with the delta power% during QS, and negative correlated with the interruption during QS. Conclusions: IH causes sleep fragmentation in both male and female mice, autonomic dysregulation and cognitive impairments in male and sham female mice, whereas it exhibited poor cognitive function in OVX female mice before IH intervention.
目 錄
英文摘要 v
中文摘要 vii
研究背景介紹 1
睡眠呼吸中止症盛行率與症狀介紹 1
睡眠呼吸中止症與認知功能之關聯性 1
睡眠呼吸中止症在性別上之差異 2
利用動物模式研究睡眠呼吸中止症的必要性 3
間歇性缺氧的動物模式 4
間歇性缺氧造成認知功能損害之潛在病理因子 5
循環變異性參數與自律神經系統 5
腦波、腦波頻譜分析及睡眠 7
自律神經系統與睡眠對認知功能之影響 8
間歇性缺氧對自律神經系統與睡眠之影響 9
停經對自律神經系統與睡眠之影響 10
研究動機與目的 11
目標 13
重要性 14
研究方法與步驟 15
動物選擇與處理 15
實驗流程 15
自製無線睡眠測量系統手術操作 15
卵巢去除手術 16
訊號收集 16
睡眠與自律神經分析方法 17
心電訊號處理 18
間歇性缺氧動物模式 19
新物體辨識測試 19
水迷宮測試 20
統計分析 20
實驗結果 21
睡眠間歇性缺氧動物模式 21
IH對公鼠、Sham與OVX母鼠心跳間距及自律神經功能之影響 21
IH對公鼠、Sham與OVX母鼠的睡眠結構及品質之影響 21
IH對公鼠、Sham與OVX母鼠在認知功能表現之影響 22
公鼠、Sham與OVX母鼠在IH期間QS期的睡眠結構與IH介入後認知功能表現之相關性 22
討論 24
本論文之重要發現 24
此生理訊號檢測動物模式與其他生理訊號檢測動物模式之差異 24
本論文IH動物模式與人類及其他IH動物模式之差異 24
IH對睡眠及自律神經功能之影響 25
睡眠IH對認知功能之影響 26
本論文之限制 27
本論文之應用價值 28
結論 29
參考文獻 30
附圖 39


圖表目錄
圖一、實驗流程圖 39
圖二、小鼠間歇性缺氧模型 40
圖三、C57BL/6J小鼠在光亮時給予8小時的間歇性缺氧 (IH) 共24小時的生理訊號原始代表圖 42
圖四、C57BL/6J小鼠有無給予每小時30次的間歇性缺氧 (IH) 共1小時的生理訊號原始代表圖 44
圖五、公鼠 (Male)、假手術 (Sham) 與去卵巢母鼠 (OVX) 的間歇性缺氧 (IH) 組與一般空氣 (RA) 組在光亮週期有無IH介入下自律神經活性參數之數值 46
圖六、公鼠 (Male)、假手術 (Sham) 與去卵巢母鼠 (OVX) 的間歇性缺氧 (IH) 組與一般空氣 (RA) 組在光亮週期有無IH介入的睡眠結構 48
圖七、公鼠 (Male)、假手術 (Sham) 與去卵巢母鼠 (OVX) 的間歇性缺氧 (IH) 組與一般空氣 (RA) 組在光亮週期有無IH介入的睡眠品質 50
圖八、公鼠 (Male)、假手術 (Sham) 與去卵巢母鼠 (OVX) 在間歇性缺氧 (IH) 介入後新物體辨識 (Novel object recognition) 之認知行為表現 51
圖九、公鼠 (Male)、假手術 (Sham) 與去卵巢母鼠 (OVX) 在間歇性缺氧 (IH) 介入後莫里斯水迷宮 (Morris water maze) 之認知行為表現 53
圖十、公鼠 (Male)、假手術 (Sham) 與去卵巢母鼠 (OVX) 在間歇性缺氧 (IH) 期間安靜睡眠 (QS) 的睡眠結構與IH介入後新物體辨識 (Novel object recognition) 認知行為表現之相關性 55
圖十一、公鼠 (Male)、假手術 (Sham) 與去卵巢母鼠 (OVX) 在間歇性缺氧 (IH) 期間安靜睡眠 (QS) 的睡眠結構與IH介入後莫里斯水迷宮 (Morris water maze) 認知行為表現之相關性 58
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