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請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/48068
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor蘇慧敏(Hui-Min Su)
dc.contributor.authorJian-Rong Linen
dc.contributor.author林建融zh_TW
dc.date.accessioned2021-06-15T06:45:31Z-
dc.date.available2011-10-05
dc.date.copyright2011-10-05
dc.date.issued2010
dc.date.submitted2011-06-27
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/48068-
dc.description.abstract婦女停經後,體內雌激素 (estrogen) 含量下降,對某些婦女而言,容易造成認知功能及情緒上的障礙。雌激素補充療法雖能有效改善停經婦女的認知功能以及焦慮、憂鬱等情緒,然而卻也增加罹患乳癌的風險。因此發展其他方式改善停經婦女的認知功能下降及情緒障礙,便是一個重要的課題。
根據研究顯示,雌激素與二十二碳六烯酸 (DHA,docosahexaenoic acid) ,均能幫助學習記憶 (learning and memory) 並改善焦慮 (anxiety) 及憂鬱(depression) 等情緒。而雌激素與DHA 在體內含量似乎有某種程度的關聯。因此本論文探討,是否能藉由補充富含DHA 的魚油,進而改善去卵巢大鼠因雌激素缺造成的學習記憶功能下降,並降低大鼠焦慮憂鬱的程度。
本研究將出生後六個月的Wistar 大鼠分成四組: 假手術組(sham operation)、去卵巢組 (ovariectomy)、假手術魚油補組、去卵巢魚油補充組。手術4 個月後開始進行行為實驗,測試大鼠空間參考記憶(spatial reference memory) 、工作記憶 (working memory) 以及焦慮、憂鬱的程度,並於鼠齡為12個月時將大鼠犧牲,分別對與學習記憶功能相關的海馬 (hippocampus) 與基底前腦 (basal forebrain) 進行組織切片染色;觀察海馬齒狀回(dentate gyrus) 神經新生 (neurogenesis),計算齒狀回體積及CA1 (cornu ammonis 1) 神經細胞面積,並計算基底前腦內膽鹼神經元 (cholinergic neuron) 的細胞數。另一方面對情緒相關的背側縫核 (dorsal raphe) 進行組織切片染色,計算其內血清素神經元(serotonergic neuron) 的細胞數。
實驗結果發現去卵巢大鼠相較於假手術大鼠,在工作記憶實驗的表現較差;給予魚油補充的大鼠相較於未補充魚油的大鼠,在空間參考記憶及工作記憶實驗的表現均較佳,且魚油補充能提高大鼠齒狀回的神經新生能力,且能減少齒狀回IV因去除卵巢後造成的體積下降。然而去除卵巢或補充魚油並未對大鼠CA1 神經細胞面積、基底前腦膽鹼神經元細胞數造成顯著的影響。而在焦慮、憂鬱程度及血清素神經元細胞數方面,去除卵巢或魚油補充也無顯著影響。
根據本論文的實驗結果,推測魚油補充可能藉由增加齒狀回神經新生的能力,避免齒狀回因去除卵巢而減少體積,進而增進大鼠的空間參考記憶及工作記憶,因此或許能利用魚油的補充,幫助缺乏雌激素之停經婦女的學習記憶能力。
zh_TW
dc.description.abstractCognitive decline and mood disorder may happen in postmenopausal women. Estrogen replacement therapy can delay cognitive decline, reduce the risk of Alzheimer’s disease and prevent mood disorder. However the therapy increases the risk of breast cancer in postmenopausal women. Therefore, it is important to find an alternative way to ameliorate the cognition and mood in postmenopausal women.
Docosahexaenoic acid (DHA, 22:6n-3) and estrogen have similar effects on neurological function. According to some studies, the concentration of DHA is related to that of estrogen in human body. The aim of this thesis is to study whether DHA enriched fish oil supplementation would prevent the decline of learning and memory and ameliorate anxiety and depression in ovariectomized rats. Six-month- old female rats were ovariectomized or sham operated and then fed n-3 fatty acids deficient diet with or without fish oil supplementation for six months. We hypothesized that ovariectomy (OVX) may decrease the ability of learning and memory, dentate gyrus volume, CA1 neuron cell size, neurogenesis in hippocampus, the number of cholinergic neurons in basal forebrain. Moreover, OVX may decrease the number of serotonergic neurons in dorsal raphe leading to anxiety and depression. While, fish oil
supplementation may retrieve back those changes.
We found that ovariectomized rats showed significantly poorer working memory than sham operated rats and fish oil supplementation rescued the memory performance. In addition, fish oil supplementation increase hippocampal neurogenesis in ovariectomized rats. Decreasing size of dentate gyrus in ovariectomized rats was ameliorated by fish oil supplementation. In CA1 neuron size and the number of cholinergic neurons, there was no significant difference between ovariectomized and sham operated rats with or without fish oil supplementation. On the other hand, there was no significant effect of OVX or fish oil supplementation on anxiety, depression and the number of serotonergic neurons.
According to our study, fish oil supplementation improves working memory and increases hippocampal neurogenesis in ovariectomized rats. These results suggest that fish oil supplementation has the benefit on learning and memory in postmenopausalwomen.
en
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dc.description.tableofcontents目錄
誌謝……………………………………………………………………Ⅰ
中文摘要………………………………………………………………Ⅲ
英文摘要……………………………………………………………..Ⅴ
目錄……………………………………………………………………Ⅶ
圖目錄………………………………………………………………ⅩⅠ
表目錄………………………………………………………………ⅩⅢ
第一章 引言……………………………………………………………1
第二章 文獻回顧……………………………………………………3
第一節 雌激素………………………………………………………3
第二節 雌激素與神經系統…………………………………………4
第三節 二十二碳六烯酸……………………………………………6
第四節 二十二碳六烯酸與神經系統………………………………7
第五節 雌激素與二十二碳六烯酸的關聯…………………………9
第三章 實驗目的與假設……………………………………………..10
第四章 材料與方法…………………………………………………..11
第一節 實驗設計……………………………………………......11
第二節 動物飼料……………………………………………………14
第三節 去卵巢手術及假手術………………………………………16
第四節 行為實驗……………………………………………………17
(一) 十字迷宮實驗…………….........…………………...17
(二) 強迫游泳實驗…..………………………………...…….18
(三) 被動閃避學習實驗..…………………...……………….19
(四) 雙向主動閃避學習實驗…..………………………………21
(五) 水迷宮實驗...……………..…………………………….22
5-1 空間參考記憶實驗...…………………………………23
5-2 工作記憶實驗...………………………………………23
第五節 BrdU 注射…...……………………………………………24
第六節 組織染色………………………..…………………………25
(一) 心臟灌流..…...………………………………………….25
1-1 藥品…………………………………………………….25
1-2 步驟…………………………………………………….26
(二) 免疫組織化學染色………..…………...……………….26
2-1 切片…………………………………………………….26
2-2 BrdU 免疫組織化學染色………………………………27
2-2.1.藥品……………………………………………….27
2-2.2.步驟……………………………………………….28
2-3 BrdU 免疫組織螢光染色………………………………29
2-4 ChAT、tryptophan hydroxylase免疫組織化學染色.30
2-4.1.藥品……………………………………………….30
2-4.2.步驟……………………………………………….31
(三) 尼氏染色………………..…………………………………32
3-1 藥品…………………………………………………….32
3-2 步驟…………………………………………………….32
第七節 細胞計數………………..………………………………..32
(一) BrdU 陽性反應細胞之計數……………………………….32
(二) ChAT 陽性反應細胞之計數……………………………….33
(三) Tryptophan hydroxylase 陽性反應細胞之計數……….34
第八節 海馬齒狀回體積的量測……………………………………34
第九節 海馬CA1 神經細胞細胞體面積的量測……………………35
第十節 統計分析……………………………………………………36
第五章 實驗結果….………………………………………………….37
第一節 子宮及體重變化……………………………………………37
第二節 水迷宮實驗..……………………………….…………….39
(一) 空間參考記憶實驗…………………………………………39
(二) 工作記憶實驗…………………….……………………….44
第三節 BrdU陽性反應細胞…………………………………………48
第四節 海馬齒狀回之體積..……………………………..………51
第五節 海馬CA1 神經細胞細胞體面積……………………………54
第六節 ChAT 陽性反應細胞……………………….………………58
第七節 十字迷宮實驗…..……………………………..…………63
第八節 強迫游泳實驗…………………………..…………………66
第九節 Tryptophan hydroxylase 陽性反應細胞……………...69
第六章 討論……………………………………………………………72
第一節 去除卵巢對大鼠學習記憶的影響……….……………….72
第二節 補充魚油對學習記憶的影響……………….…………….73
第三節 去除卵巢對海馬的影響……………………………………74
第四節 補充魚油對海馬的影響……………………………………76
第五節 去除卵巢對大鼠情緒的影響………………………………78
第六節 補充魚油對大鼠情緒的影響………………………………79
第七節 去除卵巢對不同神經傳導物質系統的影響………………80
第八節 補充魚油對不同神經傳導物質系統的影響………………81
第七章 結論……………………………………………………………83
Appendix……………………………………………………………….84
A、被動閃避學習實驗………………………………………………84
B、雙向主動閃避學習實驗…………………………………………85
C、免疫組織螢光染色………………………….………………….87
參考文獻……………………………………………………….………89
圖目錄
圖一、動物分組與實驗流程圖……………………………………….12
圖二、十字迷宮……………………………………………………….17
圖三、強迫游泳實驗之水缸………………………………………….18
圖四、Passive avoidance/active avoidance test 被、主動閃避學習系統…….............................................19
圖五、Shuttle Flex Test Chamber Model-SFLX 儀器…………..21
圖六、水迷宮………………………………………………………….22
圖七、各組大鼠子宮重量…………………………………………….37
圖八、各組大鼠於手術後體重變化………………………………….38
圖九、空間參考記憶折線圖……………………..………………….41
圖十、空間參考記憶實驗每天個別4 次訓練的逃脫時間………….42
圖十一、工作記憶折線圖…………………………………………….46
圖十二、免疫組織化學染色-背側海馬齒狀回顆粒下區BrdU陽性反應細胞…….................................................49
圖十三、大鼠背側海馬齒狀回顆粒下區BrdU 陽性反應細胞數……50
圖十四、尼氏染色-海馬齒狀回………………………………………52
圖十五、大鼠海馬齒狀回體積………………………………...……53
圖十六、尼氏染色-海馬CA1 神經元…………………………………55
圖十七、大鼠海馬CA1 神經細胞細胞體面積..…………………….56
圖十八、免疫組織化學染色-Basal nucleus of Myenert 的ChAT陽性反應細胞….............................................59
圖十九、大鼠basal nucleus of Myenert 內ChAT 陽性反應細胞數…………...............................................60
圖二十、免疫組織化學染色-HDB 的ChAT 陽性反應細胞………….61
圖二十一、大鼠HDB內ChAT陽性反應細胞數………………………..62
圖二十二、十字迷宮實驗結果-大鼠探索open arms 的時間總和…64
圖二十三、十字迷宮實驗結果-大鼠進入open arms 的次數………64
圖二十四、十字迷宮實驗結果-大鼠進出open arms 與closed arms 的次數總和….............................................65
圖二十五、強迫游泳實驗結果…………………………….........67
圖二十六、免疫組織化學染色-背側縫核tryptophan hydroxylase 陽性反應細胞…...........................................70
圖二十七、大鼠背側縫核內tryptophan hydroxylase 陽性反應細胞數…………….............................................71
圖A1、被動閃避學習實驗結果……………………………………….84
圖A2、雙向主動閃避學習實驗結果………………………………….86
圖A3-1、SD組OD組大鼠BrdU、NeuN免疫組織螢光染色….…..……87
圖A3-2、SDF組ODF組大鼠BrdU、NeuN免疫組織螢光染色……...…88
表目錄
表一、自製缺乏n-3 脂肪酸飼料之成分組成…………………..….14
表二、自製n-3 脂肪酸缺乏飼料及餵食魚油之脂肪酸組成…..….15
表三、空間參考記憶實驗統計分析結果…..…………..………….43
表四、工作記憶實驗統計分析結果……………………..………….47
表五、海馬染色統計分析…………………………………………….57
表六、ChAT 陽性反應細胞計數統計分析…………………………..62
表七、十字迷宮與強迫游泳實驗統計分析………………………….68
表八、Tryptophan hydroxylase 陽性反應細胞計數統計分析…..71
dc.language.isozh-TW
dc.title探討魚油補充對去卵巢大鼠空間學習記憶及焦慮憂鬱之影響zh_TW
dc.titleEffects of fish oil supplementation on spatial learning,
anxiety and depression in ovariectomized rats
en
dc.typeThesis
dc.date.schoolyear99-2
dc.description.degree碩士
dc.contributor.oralexamcommittee蔡元奮(Yuan-Feen Tsai),盧國賢(Kuo-Shyan Lu),黃青真(Ching-jang Huang)
dc.subject.keyword魚油,去卵巢大鼠,空間學習記憶,焦慮,憂鬱,海馬,神經新生,zh_TW
dc.subject.keywordfish oil,ovariectomized rats,spatial learning,anxiety,depression,hippocampus,neurogenesis,en
dc.relation.page100
dc.rights.note有償授權
dc.date.accepted2011-06-27
dc.contributor.author-college醫學院zh_TW
dc.contributor.author-dept生理學研究所zh_TW
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