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研究生:黃雅芬
研究生(外文):Ya-Fen Huang
論文名稱:蜜環菌醱酵物增長戊巴比妥鈉誘導小鼠睡眠效果
論文名稱(外文):The enhancement of pentobarbital sodium-induced hypnosis in mice by fermentation products of Armillariella mellea
指導教授:李順來李順來引用關係
指導教授(外文):Shun-Lai Li
學位類別:碩士
校院名稱:南台科技大學
系所名稱:生物科技系
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
中文關鍵詞:蜜環菌睡眠
外文關鍵詞:Armillariella mellea
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蜜環菌(Armillariella mellea,AM),分佈於全球各地區,為口磨科植物,也是偶見木棲腐生或木棲寄生的野菇之一種,為食療皆可。本實驗利用蜜環菌與中藥材天麻及酸棗仁進行共同醱酵,所得醱酵物餵食小鼠七日後進行睡眠實驗測試,以戊巴比妥鈉(pentobarbital sodium) 60mg/kg誘導小鼠入睡,並觀察小鼠入睡時間與睡眠時間是否改變。入睡時間是從注射戊巴比妥鈉後到失去翻正反射動作期間,而睡眠時間則是從失去到恢復反射動作期間稱之。
利用基質一的蜜環菌天麻醱酵物餵食組,小鼠比未餵食組別小鼠的睡眠時間延長70%,餵食蜜環菌酸棗仁發酵物也可延長48%睡眠時間。而餵食不同劑量蜜環菌天麻及蜜環菌酸棗仁醱酵物小鼠入睡時間較低於未餵食組別78%到88%。 單純餵食基質二醱酵物組別能縮短入睡時間76%,此結果証實經由基質一的中草藥蜜環菌醱酵及單純基質二蜜環菌醱酵物產品對幫助睡眠具有相當顯著的效果。
故選用基質一與蜜環菌天麻醱酵物10 mg/20g、20 mg/20g、與酸棗仁醱酵物2 mg/20g及基質二與蜜環菌醱酵物20 mg/20g,此四個組別經初篩後具有顯著性的縮短入睡時間或延長睡眠時間效果做為實驗組別,並進一步測定重覆實驗、次濃度誘導小鼠入睡和抗耐藥性實驗。重複進行睡眠實驗結果與次濃度45mg/kg注射戊巴比妥鈉誘導小鼠入睡實驗中,經由餵食蜜環菌醱酵物組別是入睡時間上有明顯縮短效果,在睡眠時間上並未出現顯著影響;另外在抗耐藥性實驗上,連續注射初期七日,各組入睡時間縮短效果明顯,睡眠延長效果不佳;連續注射至後期,睡眠效果才有明顯的延長功效表現。
將進行餵食一週與四週後小鼠進行其大腦皮質Na+-K+-ATPase活性測試,餵食一週後之實驗組別僅餵食基質一天麻醱酵物有活性下降趨勢,而餵食基質一酸棗仁醱酵組及基質二醱酵組別其活性則增高;餵食四週後小鼠Na+-K+-ATPase活性,在四組餵食實驗組別之活性皆顯著高於控制組,與其他前人研究相反。故醱酵物影響睡眠效果機制點應可往Na+-K+-ATPase相關機制做探討。
Armillariella mellea is one of the Tricholomataceae mushrooms that spread worldwide and can be used as food and medicine. This study uses the fermentation products of Armillariella mellea with the addition of Gastrodia elata Blume or Ziziphus spinosa to explore the phenomenon of hypnosis in mice with the pentobarbital sodium(60mg/kg)-induced .Mice were fed with fermentation products twice daily in a 7 days period prior to hypnosis test. The tests are going to see whether the sleeping onset time and duration time would be changed or not in the pentobarbital-induced mice. The onset time is the duration of time from the injection of pentobarbital sodium till the loss of righting reflex ability; the sleeping duration time is the period of time from loss the ability of righting reflex to its recovery.
The study showed that the sleeping duration time induced by pentobarbital sodium(60mg/kg)was 70% longer than that of control by treating with the fermentation product of Gastrodia elata Blume and substance1 fermented by Armillariella mellea. Mice Treated with product of Ziziphus spinosa and substance1 fermented by Armillariella mellea prolonged 48% of the sleeping duration time than that of control. The onset time by treating with the fermentation products of Armillariella mellea with Gastrodia elata Blume or Ziziphus spinosa was 78% to 88% shorter than that untreated. But those treated with product of chinese herbs and substance2 fermented by Armillariella mellea did not affect hypnosis in mice.
The onset time and sleeping duration time that were treated with the fermentation products of Armillariella mellea with Gastrodia elata Blume and substance1 in 10 mg/20g、 20 mg/20g, Ziziphus spinosa and substance1 in 2mg/20g and substance2 in 20mg/20g as test groups are different from control(p<0.05).The aforementioned four test groups were again to evaluate the hyponosis phenomena of mice under subhypnotic dosage and their anti-tolerance effect. The sleeping onset time of mice treated with fermentation products by subhypnotic dosage (45mg/kg) and 60mg/kg of pentobarbital are shorter than control, but sleeping duration time have not been affected. The onset time is shorter than control for mice treated with fermentations products with continuous injection of pentobarbital sodium (60mg/kg) in 7 primary days in anti-tolerance evaluation, and the prolonged sleeping duration time did not appeared until the 7th day of fermentation product treatment.
The Na+-K+ ATPase activities in cerebral cortex of mice were inhibited after treated with fermentation products of Armillariella mellea with Gastrodia elata Blume and substance1 in 10 mg/20g、 20 mg/20g for 1 week. However the activities were increased for mice treated with fermentation products of Armillariella mellea with Ziziphus spinosa and substance1 in 2mg/20g and with substance2 in 20mg/20g. The ATPase activities in mice treated with fermentation products for 4 weeks, however, were significantly higher than control. In conclusion, the mechanism of hyponotic effects between Na+-K+ ATPase activities and the fermentation products of Armillariella melle should be further investigated.
目次
摘要…………………………………………………………………..…….………….…i
英文摘要………………………………………………………………………………...ii
目次………………………………………………………………………….…….……iv
表目錄………………………………………………………………………………......vi
圖目錄……………………………………………………………………….…………vii
第一章 前言…………………………………………………………………….………1
第二章 文獻整理……………………………………………………………….………3
2.1 蜜環菌藥理研究…………………………………….………………..….………4
2.2 天麻(Gastrodia elata Blume)介紹……………….………………………....……5
2.3 酸棗仁(Ziziphus spinosa)介紹………………………….………………….…….6
2.4 造成失眠的原因…………………………………………………...………..…...7
2.5 與睡眠相關分子機制………………….……………………...……………..…10
2.6 Na+-k+ATPase活性與機制………………………………...………….……....14
2.7 戊巴比妥鈉的介紹…………………………….…………………….…………20
第三章 實驗架構………………………………………..………………………….…24
第四章 材料與方法
4.1 菌種來源…………………………...……………..…………………………….25
4.2 實驗設備及藥品……………………...…………..…………………………….25
4.3 液態醱酵培養菌絲體………………...………..……………………………….26
4.4 多醣含量測定………………………...………..……………………………….27
4.5 改善睡眠功能實驗…………………………….……………………………….29
4.6 Na+-k+ATPase 活性測試………….………….…...…………………………..30
4.7 統計分析……………………..……………...………………………………….33
第五章 結果與討論
5.1 蜜環菌生長曲線測定…………..…….………………………………...…........34
5.2 蜜環菌多醣測定……………...………………………………………………...36
5.3 蜜環菌醱酵物對利用戊巴比妥鈉所誘導睡眠效果之篩選………….….. …..37
5.4 蜜環菌醱酵物改善以戊巴比妥鈉誘導睡眠效果之重覆實驗結果…..………45
5.5 次濃度戊巴比妥鈉誘導小鼠睡眠效果探討…..…………………………........48
5.6 抗耐藥性實驗…………………………………………………………………..50
5.7 Na+-K+ ATPase活性測試..……….……………………………………………60
第六章 結論…………………………………….……………………………………62
參考文獻………………………………………………………………………….……65
附件……………………………………………………………………………….……68
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