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研究生:黃郁婷
研究生(外文):Huang,Yu-ting
論文名稱:探討北冬蟲夏草之培養條件及其抗疲勞作用
論文名稱(外文):Study on the cultivated conditions and the anti-fatigue effect ofCordyceps militaris
指導教授:張惠香
口試委員:王志傑、李築宜
口試日期:2013-01-14
學位類別:碩士
校院名稱:大仁科技大學
系所名稱:生物科技研究所
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:2013
畢業學年度:101
語文別:中文
論文頁數:93
中文關鍵詞:北冬蟲夏草、海洋深層水、抗氧化、抗疲勞、耐久性運動
外文關鍵詞:Cordyceps militaris、Deep ocean water、Antioxidative、antifatigue、endurance exercise
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北冬蟲夏草(Cordyceps militaris)是由感染昆蟲幼蟲或蛹體所形成之藥材。北冬蟲夏草之功能性成分有蟲草素、蟲草酸、核苷類、蟲草多醣等,其在人體醫療、保健方面有重要作用的應用。海洋深層水中富含豐富無機鹽及離子性礦物元素,其中礦物元素無法由人體自行合成必須由外在補充的方式取得,而微量元素Ca2+、Mg2+離子能提升酵母菌生長或麴菌及樟芝菌活性功能。結果顯示,比較液態培養於市售不同廠牌海洋深層水之北冬蟲夏草,其中在菌絲體質量並無明顯差異。而在總多醣含量分析方面以 Taiwan Yes 100% (TY100) 深命力海洋深層水之北冬蟲夏草含量為較高。因此,以此為後續研究選用的水源。利用不同濃度 DOW 濃縮液去探討添加過多離子對於北冬蟲夏草影響,結果得知原本一倍 DOW 的濃度就能使總多醣生成增加。此外,我們另外利用一倍DOW濃度中單一離子去做後續實驗。結果發現去離子培養基額外添加 Mg2+、Ca2+、K+ 離子能增加蟲草素的含量,而 Mg2+ 離子能提升總多醣生成。在固態培養方面,以TY 100之50%濃度含量培養下所得之子實體的生物質量較高,而在75% 濃度的 TY 培養下,添加豆粕粉(SM)則可提升蟲草素及總多醣的含量。在抗氧化能力部分在10 mg/mL 濃度下 DPPH 清除自由基能力跟 Vitamin C 清除效果一樣。在不同光質照射下北冬蟲夏草於固體培養結果以綠光有助於菌體量及總多醣之生成。另外餵食北冬蟲夏草市售產品進行動物實驗模式依照衛生署公佈之健康食品抗疲勞評估方法做檢測。利用老鼠游泳耐力試驗(Endurance Exercise)作為模式,藉以測試運動前、後之血液中血糖、乳酸、尿素氮等濃度之變化,來探討北蟲草對延緩運動期間疲勞發生之能力影響。本研究以10週齡SD大鼠50隻共分為五組,除控制組外另分一組正控制組及三組分別每天餵食0.5、1及2.5倍之北蟲草產品之實驗組。經28天的灌食後,結果顯示北蟲草實驗組比控制組有較長游泳時間及較高的肝醣濃度。另外實驗組血中尿素氮及乳酸濃度的上升率也都比控制組低。因此由研究顯示,老鼠攝食北蟲草產品可以提高抗疲勞情形並對耐久性之運動具有改善生理功能之作用。
Cordyceps militaris is a herbal medicine grew on infected insect larvae or pupae. The Cordyceps militaris contains functional ingredients including cordycepin, cordyceps acid, nucleoside and Cordyceps polysaccharide, and is an important medicine in health and medicare. The deep ocean water(DOW), which contains rich ionic minerals and trace elements that could not be synthesized by human body and could only be supplemented by intake, and also contains trace elements Mg2+ and Ca2+ ions, is suitable for promoting yeast growth or for culturing the mycelia of Aspergillus and Antrodia camphorate. The results of this study show that the Cordyceps militaris, liquid-cultivated in deep ocean water obtained from different manufacturers, showed no significant difference in quality and content of the mycelium. In the aspects of total polysaccharide content, the Cordyceps militaris cultivated in the Taiwan Yes 100% (TY100) deep life force deep ocean water is found to be relatively higher, thus such water is chosen as clultivation liquid in the follow-up study. Concentrated DOW, in different concentrations, was evaluated for the effect of adding excess ions on Cordyceps militaris cultivation. The preliminary result indicated that the use of DOW in the original concentration can increase the total polysaccharide content significantly. We further used individual Mg2+, Ca2+, K+ ions at concentration equivalent to that in DOW of the original concentration in the follow-up experiments. The results showed that the use of deionized medium with added Mg2+, Ca2+ and K+ ions can resulted in increased cordycepin content, while Mg2+ ions can improve the total polysaccharides content. In the solid-state culture, the use of TY 100 at 50% concentrations yielded fruiting bodies with greater biomass content. Meanwhile, the use of TY in 75% concentration, with added soybean meal (SM), raised the content of cordycepin and total polysaccharide. Comparing irradiation effect under different light quality, the green light afforded the greatest bacteroid and total polysaccharide content in Cordyceps militaris cultivated in solid culture. Concentration of 10 mg / mL in the antioxidant capacity DPPH scavenging effect of free radical with Vitamin C. Comparing irradiation effect under different light quality, the green light afforded the greatest bacteroid and total polysaccharide content in Cordyceps militaris cultivated in solid culture. Another feeding of Cordyceps militaris commercially available products animal model anti-fatigue assessment methods to do testing in accordance with the Department of Health announced the health food was adopted, and mice swimming endurance exercise was performed. Cordyceps militaris-treated mice was evaluated for change in blood glucose, blood lactate, blood urea nitrogen and glycogen concentration before and after exercises, in order to investigate the efficacy of Cordyceps militaris in fatigue reduction after exercise. Fifty male adult SD rats of 10 weeks of age were studied by dividing them into five groups. Except for the control group, the positive control group and the rest o three groups were all fed with 0.5, 1 and 5 fold Cordyceps militaris products in the experiment. After 28 days of feeding, the result showed that the experimental groups exhibited longer exercise time and higher glycogen concentration than that of the control group. Moreover, blood urea nitrogen and blood lactic acid content of the experimental groups was also found to be significantly lower than that of the control group. Besides, the result suggested that the feeding of Cordyceps militaris products could reduce the degree of fatigue and improve the physiological condition of the tested rats.
中文摘要..................................................................................................I
英文摘要................................................................................................II
致謝............................................................................................... .......IV
目錄.......................................................................................................V
圖次....................................................................................................VIII
表次..................................................................................................... IX
壹、 前言........................................................................................................1
貳、 文獻回顧................................................................................................3
一、 北冬蟲夏草簡介…......................................................................3
(一) 北冬蟲夏草分類.......................................................................3
(二) 北冬蟲夏草之生活史...............................................................3
二、 北冬蟲夏草藥理作用...................................................................8
三、 北冬蟲夏草成份之研究...............................................................8
(一) 一般化學成份...........................................................................8
(二) 機能性成份...............................................................................8
四、 北冬蟲草相關研究....................................................................10
(一) 在維護肝臟功能方面.............................................................10
(二) 在維護腎臟功能方面.............................................................10
(三) 在心血管系統維護方面.........................................................11
(四) 抗腫瘤作用.............................................................................11
(五) 抗氧化作用.............................................................................11
(六) 抗疲勞作用.............................................................................12
五、 北冬蟲夏草培養條件................................................................13
(一) 液態培養.................................................................................13
(二) 固態培養.................................................................................14
六、 海洋深層水................................................................................15
(一) 海洋深層水介紹.....................................................................15
(二) 海洋深層水特性.....................................................................16
(三) 海洋深層水之微量礦物質功能.............................................17
(四) 海洋深層水保健之功效.........................................................18
(五) 海洋深層水於微生物發酵之相關應用................................19
七、 研究動機及目的........................................................................19
參、 北冬蟲夏草之培養條件探討..............................................................22
一、 前言............................................................................................22
二、 材料與方法................................................................................22
(一) 供試菌珠來源及培養...................................................22
(二) 供試材料.......................................................................23
(三) 供試儀器設備...............................................................24
(四) 液態培養方法與條件...................................................27
(五) 固態培養方法與條件...................................................27
(六) 北冬蟲夏草功能性成份分析.......................................28
(七) 北冬蟲夏草之抗氧化能力之測定...............................31
(八) 統計分析.......................................................................33
三、 結果............................................................................................34
(一) 海洋深層水對北冬蟲夏草菌株於液態培養下代謝成份之影響............................................................................................34
(二) 北冬蟲夏草於固體培養海洋深層水之菌體生成量及代謝成份之影響................................................................................46
(三) 北冬蟲夏草抗氧化分析…....................................................53
(四) 北冬蟲夏草於不同光質固體培養下菌體重及成份分析之影響............................................................................................57
四、 討論...........................................................................................59
(一) 北冬蟲夏草於海洋深層水(DOW)液態培養成份之影響............................................................................................59
(二) 北冬蟲夏草於固態培養不同條件下成份之影響............................................................................................59
(三) 北冬蟲夏草抗氧化分析........................................................60
(四) 北冬蟲夏草固態培養於不同光質照射下成份之影響........61
五、 結論............................................................................................62
肆、 北冬蟲夏草之其抗疲勞作用..............................................................63
一、前言..............................................................................................63
二、材料與方法..................................................................................64
(一) 研究材料................................................................................64
(二) 動物器具相關設備................................................................65
(三) 研究設計................................................................................65
(四) 實驗方法................................................................................67
(五) 統計分析................................................................................69
三、結果............................................................................................70
(一) 實驗動物體重與平均飼料攝食量之變化................................70
(二) 北冬蟲夏草產品之游泳耐力試驗...........................................70
(三) 北冬蟲夏草產品對血尿氮濃度之影響...................................70
(四) 北冬蟲夏草產品對血乳酸濃度之影響...................................70
(五) 北冬蟲夏草產品對肝醣濃度之影響.......................................71
(六) 北冬蟲夏草產品大鼠之血液學變化.......................................71
四、討論.............................................................................................79
五、結果.............................................................................................79
伍、 總討論與總結論.................................................................................80
陸、 參考文獻.............................................................................................83


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