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研究生:朱桓儀
研究生(外文):Huan-yi Chu
論文名稱:綠藻對老化促進小鼠抗氧化狀態及學習記憶力之相關研究
論文名稱(外文):Effect of Chlorella on antioxidative status and learning memory ability in senescence accelerated mice
指導教授:王銘富王銘富引用關係
指導教授(外文):Ming-Fu Wang
學位類別:碩士
校院名稱:靜宜大學
系所名稱:食品營養研究所
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2008/01/
畢業學年度:96
語文別:中文
論文頁數:111
中文關鍵詞:老化促進小鼠抗氧化學習記憶綠藻
外文關鍵詞:antioxidative statussenescence-accelerated prone 8 mice.learning and memorychlorella
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研究顯示綠藻(Chlorella)具抗氧化、降血脂、抗腫瘤及降血壓等功效。本研究目的為探討給予不同形式之綠藻對老化促進小鼠 (senescence accelerated mice; SAMP8)抗氧化狀態及學習記憶能力之影響。實驗以三及六月齡之雄性、雌性SAMP8系小鼠為對象,隨機分為對照組 (Control組) 、商業化破壁綠藻組(MP組) 、商業化綠藻組(GP組) 、物理性球磨破壁綠藻組(BM組) 共4組,每組各15隻。綠藻實驗組之飼料含2.92g/kg綠藻,採自由攝食方式給予;實驗為期12週,期間紀錄小鼠攝食量、體重變化、活動量測試及老化指數評估;並以單次被動迴避試驗及主動迴避試驗評估小鼠學習記憶能力。犧牲後進行血液生化分析,檢測肝臟超氧歧化酶(SOD)、觸酶(CAT)及麩胱苷肽過氧化酶(GPx)之活性,另分析肝中丙二醛(MDA)之含量,並觀察腦部類澱粉蛋白(Aβ)沉積。結果顯示:實驗各組攝食量、體重變化量及活動量,各組之間並無顯著差異。在老化指數評估方面,3與6月齡雄性及雌性小鼠綠藻實驗組之老化總分皆顯著低於對照組(P<0.05)。於學習記憶方面,主動迴避試驗實驗顯示綠藻組均較對照組有較佳的學習記憶能力;且單次被動迴避試驗結果顯示,綠藻組於訓練後24小時可增加滯留時間,其中又以三月齡小鼠效果為佳,顯著高於對照組(P<0.05)。實驗綠藻組皆能顯著降低三及六月齡之雄性、雌性SAMP8系小鼠肝中MDA含量(P<0.05),且SOD與CAT活性有增加之趨勢。此外綠藻可顯著降低三月齡小鼠之腦部類澱粉蛋白的沉積(P<0.05),六月齡小鼠方面,於雄性小鼠有顯著差異,但雌性小鼠則無顯著影響。綜合上述結果,不同形式之綠藻皆能提升抗氧化狀態及改善學習記憶能力,但在本實驗中,不同形式之綠藻並不影響其抗氧化酵素活性,因此何種類型之綠藻能有最佳的抗氧化效果,仍需做再進一步探討。
It was demonstrated that chlorella has the effect of antioxidation , antitumor, lowering the blood fat and reducing blood pressure. The purpose of this study was to investigate the effect of different types of chlorella on learning and memory ability and antioxidative status in senescence-accelerated mice. 3 and 6-month-old senescence-accelerated male and female mice were divided into four groups (n=15 per group) : control group, commercial mechanical chlorella powder (MP) , commercial granule chlorella powder (GP) , ball milling chlorella powder (BM) .The ideal diet of experimental group contained chlorella 2.92g/kg fed food for mice. During experimental period, body weight, food intake, aging score and open flied activity were recorded for each group of mice. Single-trail passive avoidance test and active shuttle avoidance test were performed after 12 weeks of feeding. The biochemical parameters of serum was analyzed after the mice sacrificed. The antioxidative enzyme activities such as superoxide dismutase (SOD) , catalase (CAT) , glutathione peroxidase (GPx) and MDA concentrations were estimated in the liver. Furthmore, the β-amyloid of brain was observed. The results showed that there were no significant differences in the food intake, body weight and locomotion among four groups. The aging score of experimental groups were significantly lower than the control group in 3 and 6-month-old senescence accelerated male and female mice(P&lt;0.05). In learning and memory test, experimental groups of 3 and 6-month-old senescence accelerated male and female mice had better improvement in active shuttle avoidance test, and single-trail passive avoidance test showed that 3-month-old senescence accelerated male and female mice significantly were better than the control group (P&lt;0.05) .The SOD and CAT activities of experimental groups of 3 and 6-month-old senescence accelerated male and female mice were increased, and the MDA concentrations in the liver significantly lower than the control group. The β-amyloid protein deposition of brain was significantly reduced in 3-month-old senescence accelerated mice (P&lt;0.05) ,but there was not significantly different in 6-month-old senescence accelerated mice. It is concluded that the supplement of chlorella could promote the antioxidative status and improve learning and memory ability. In this study, there was no significant difference between the experimental groups, which were made from different types. And that which type of chlorella possesses the best antioxidant capacity will further investigate.
中文摘要------------------------------------------------------------------------I
英文摘要-------------------------------------------------------------------------III
目錄-------------------------------------------------------------------------------V
表目錄----------------------------------------------------------------------------X
圖目錄----------------------------------------------------------------------------XI
第一章 前言 --------------------------------------------------------------------1
第二章 文獻回顧 --------------------------------------------------------------4
第一節 綠藻簡介 ---------------------------------------------------------4
壹、 綠藻的生理功能 ----------------------------------------------------4
一、 抗氧化作用 -----------------------------------------------------6
二、抗腫瘤作用 ------------------------------------------------------6
三、 調節血脂 --------------------------------------------------------7
四、降低血壓 ---------------------------------------------------------8
五、免疫作用 --------------------------------------------------------8
六、幫助體內排除戴奧辛毒物 ------------------------------------9
七、安全性評估 -----------------------------------------------------9
第二節 自由基對生物體的影響 ---------------------------------------10
壹、自由基與活性氧之簡介 --------------------------------------------10
貳、活性氧與自由基之來源 --------------------------------------------10
第三節 自由基與老化 --------------------------------------------------14
第四節 氧化傷害對老化疾病之相關性 ------------------------------16
第五節 氧化壓力與腦部之相關性--------------------------------------17
第六節 學習記憶測試常用動物模式與實驗法 ---------------------19
壹、 學習記憶的形成 --------------------------------------------------19
貳、 記憶形成的機制 ---------------------------------------------20
参、 學習記憶實驗方法 ------------------------------------------------20
第七節 實驗動物模式 ----------------------------------------------23
一、老化促進小鼠的來源 ---------------------------------------------23
二、老化促進小鼠之老化特徵及抗氧化狀態 ---------------------23
三、老化促進小鼠之病理特徵 ---------------------------------------25
四、老化促進小鼠對學習記憶影響 ---------------------------------26
五、老化指數系統評估 ------------------------------------------------27
第八節 生物體內抗氧化防禦系統 ------------------------------------30
壹、 酵素型抗氧化劑 --------------------------------------------------30
貳、 非酵素型抗氧化劑 -----------------------------------------------31
一、維生素 E ----------------------------------------------------------31
二、維生素 C ----------------------------------------------------------31
三、酚類化合物 ---------------------------------------------------------30
四、β-胡蘿蔔素 -------------------------------------------------------32
第九節、脂質過氧化物 ----------------------------------------------------32
第三章 材料與方法 ----------------------------------------------------------34
第一節 實驗動物 ---------------------------------------------------------34
第二節 實驗設計 ---------------------------------------------------35
第三節 實驗方法與步驟 ------------------------------------------------37
壹、實驗流程 ----------------------------------------------------------38
貳、活動量測試 -------------------------------------------------------39
叁、老化指數 ----------------------------------------------------------39
肆、學習記憶能力測試 ----------------------------------------------40
ㄧ、主動迴避試驗 -------------------------------------------------40
二、被動迴避試驗 -------------------------------------------------41
伍、血液生化分析 -------------------------------------------------45
陸、抗氧化能力分析-------------------------------------------------45
ㄧ、超氧歧化酶 ----------------------------------------------------45
二、觸酶 -------------------------------------------------------------47
三、麩胱苷肽過氧化酶 -------------------------------------------48
四、丙二醛 ----------------------------------------------------------49
柒、 腦部病理組織切片 ---------------------------------------------51
ㄧ、試藥製備 -------------------------------------------------------51
二、組織切片製作過程 -------------------------------------------53
三、腦組織病理評估 ----------------------------------------------59
第四節 統計分析 --------------------------------------------------------61
第四章 結果與討論 ---------------------------------------------------------62
第一節 體重及攝食量變化 --------------------------------------------62
第二節 活動量 -----------------------------------------------------------62
第三節 老化指數 --------------------------------------------------------67
第四節 學習與記憶能力 -----------------------------------------------72
壹、主動迴避試驗 ------------------------------------------------------72
貳、單次被動迴避試驗 ------------------------------------------------77
第五節 器官重量 ------------------------------------------------------82
第六節 血液生化學分析 ----------------------------------------------82
第七節 抗氧化酵素活性及脂質過氧化物 -------------------------88
壹、 超氧歧化酶活性 ----------------------------------------------- 88
貳、 觸酶活性 ------------------------------------------------------88
叁、 麩胱苷肽過氧化酶活性 -----------------------------------------88
肆、丙二醛含量測定 ---------------------------------------------------88
第八節 腦部-類澱粉蛋白沉積 ------------------------------------98
第五章 結論 -----------------------------------------------------------------102
參考文獻 -----------------------------------------------------------------------103
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