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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 葉安義 | |
| dc.contributor.author | Yi-Hsien Chiang | en |
| dc.contributor.author | 江怡嫻 | zh_TW |
| dc.date.accessioned | 2021-06-08T00:55:13Z | - |
| dc.date.copyright | 2015-03-16 | |
| dc.date.issued | 2015 | |
| dc.date.submitted | 2015-02-16 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18216 | - |
| dc.description.abstract | 靈芝是一種使用歷史悠久，具有生理功能的藥用真菌類，傳統上以熱水萃取方式獲得靈芝茶或將子實體乾燥研磨成粉末後使用。本實驗以介質研磨方式製備奈米/次微米靈芝懸浮液，與熱水萃取物之成分以及功能性進行比較，並以細胞毒性試驗以及致突變性試驗，初步確認奈米/次微米靈芝懸浮液之安全性。靈芝子實體在 3小時的介質研磨處理過程中，菌絲的中空結構被撕裂為片狀後，不斷與研磨介質產生摩擦、碰撞，變成碎片，最後變為微小碎片。研磨的產品中，大部分的懸浮粒子之粒徑小於1 μm，且有 62 % 的粒子小於0.1 μm；以掃描式電子顯微鏡 (SEM) 觀察，也確認了樣品的粒徑為奈米/次微米等級。濾除渣質後，奈米/次微米靈芝懸浮液收率為21.5%，靈芝熱水萃取物則為15.4%，且介質研磨樣品中多醣、蛋白質與1,3-β-glucan (3.5倍) 等物質含量也較熱水萃取組高，顯示以介質研磨處理方式萃取效率較佳。以0.45 μm濾膜除菌的奈米/次微米靈芝懸浮液 (S-supernatant) 與靈芝熱水萃取物 (S-extract) 並不會增加沙門氏菌的突變率，也不具有抑制突變的效果。細胞毒性試驗則顯示，樣品對小鼠消化道、肝臟與腎臟的初代細胞不具有毒性，且會刺激細胞生長，提升細胞存活率。奈米/次微米靈芝懸浮液的多醣萃取物中所含的1,3-β-glucan較熱水萃取組多出約1.7倍，但分支少，分子量較低；蛋白質萃取物中有部分蛋白質分子與冷水萃取物蛋白質的分子量相似，但多醣含量較高，熱水萃取組蛋白質的結構則大多已被高溫破壞。免疫活性測定顯示奈米/次微米靈芝與熱水萃取物，以及兩者的粗多醣與粗蛋白萃取物，皆會刺激小鼠腹腔巨噬細胞分泌細胞激素，其中以s-supernatant的效果最強。介質研磨處理方法所製備的三組樣品傾向增加脾臟細胞的Th1細胞激素分泌，並抑制Th2細胞激素，動物實驗也呈現類似結果；熱水萃取方式處理之三組樣品，則傾向同時促進Th1與Th2細胞激素的分泌。本研究初步判定以介質研磨方式製備奈米/次微米靈芝懸浮液，所獲得的萃取物不具有細胞毒性與致突變性，且活性成分與功效不同於傳統熱水萃取物，活性成分結構與免疫調節效果之相關性，則有待更進一步的分析。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-06-08T00:55:13Z (GMT). No. of bitstreams: 1 ntu-104-D96641003-1.pdf: 3532745 bytes, checksum: 2955bea8d310cc6958959edaee075d61 (MD5) Previous issue date: 2015 | en |
| dc.description.tableofcontents | 目錄
摘要 I ABSTRACT III 目錄 V 圖目錄 VIII 壹、前言 1 貳、文獻回顧 2 一、靈芝 2 二、靈芝的生理活性與成分 3 三、奈米科技與濕式介質研磨 6 四、奈米/次微米粒子之安全性 10 五、免疫系統 12 參、實驗架構 15 一、樣品製備 15 二、各種樣品之分析項目 16 三、免疫調節試驗 17 肆、材料與方法 18 一、材料與預處理 18 1. 松杉靈芝預處理 18 2. 熱水萃取方法 18 3. 介質研磨方法 18 二、基本成分分析 19 三、粒徑分析 21 四、顯微觀察 22 五、成分與有效物質分析 22 1固形物含量測定 22 2. 醣含量測定 23 3. 蛋白質含量測定 23 4. 1, 3- β- D-glucan 含量測定 23 5. 三萜類分析 24 6. 可溶性多醣分析 25 7. 蛋白質分析 28 六、奈米/次微米靈芝懸浮液之致突變性試驗與細胞毒性試驗 31 1. 樣品處理與製備 32 2. 奈米/次微米靈芝懸浮液之致突變性試驗 32 3. 細胞毒性試驗 34 七、奈米/次微米靈芝懸浮液免疫調節能力測定 35 1. 樣品 35 2. 小鼠巨噬細胞株 (RAW 264.7) 存活率測定 36 3. 細胞激素TNF-α分泌量之測定 36 4. BALB/c小鼠腹腔細胞取得 37 5. BALB/c小鼠腹腔細胞培養 37 6. BALB/c小鼠脾臟細胞取得 38 7. BALB/c小鼠脾臟細胞培養 38 8. 細胞激素分泌量之測定 39 9. 奈米/次微米靈芝懸浮液對Balb/c鼠體內非特異性免疫反應之影響 39 八、數據統計分析 44 伍、結果與討論 45 一、靈芝子實體顯微觀察與基本成分分析 45 二、粒徑分析與顯微觀察 48 三、松杉靈芝萃取物組成分與有效物質分析 56 四、三萜類含量與組成分析 59 五、致突變性實驗 (安姆試驗) 61 六、細胞毒性試驗 68 七、可溶性多醣萃取物分析 72 1. 單醣組成 72 2. 可溶性1, 3-β-D-葡聚醣與其分支度測定 76 3. 分子量與1, 3-β-D-葡聚醣分佈 79 八、蛋白質萃取與分析 81 1. 分子篩管柱分離 81 2. DEAE管柱分離 83 3. 聚丙烯醯胺凝膠電泳 (SDS page) 85 九、樣品對小鼠巨噬細胞株存活率與細胞激素分泌之影響 91 1. 細胞株存活率 91 2. 細胞激素TNF-α分泌量之測定 95 十、樣品對小鼠腹腔巨噬細胞的細胞激素分泌之影響 96 十一、樣品對小鼠初代脾臟細胞的細胞激素分泌之影響 104 十二、奈米/次微米靈芝懸浮液對Balb/c鼠體內非特異性免疫反應之影響 110 1. 動物飼養 110 2. 血液中吞噬細胞活性之分析 110 3. 脾臟免疫細胞表面抗原分析 111 4. 脾臟細胞代謝活性分析 115 5. 細胞激素分泌量之測定 115 陸、 結論 120 柒、 參考文獻 121 | |
| dc.language.iso | zh-TW | |
| dc.title | 奈米/次微米靈芝懸浮液與其功能性成分對免疫調節之影響 | zh_TW |
| dc.title | Immunomodulatory Effect of the Nano/submicron Ganoderma tsugae and the Bioactive Components | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 103-1 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 蔣丙煌,孫璐西,盧訓,張永和,黃青真 | |
| dc.subject.keyword | 介質研磨,靈芝,細胞毒性,致突變性,免疫, | zh_TW |
| dc.subject.keyword | Ganoderma tsugae,media milling,Immunomodulation,mutageenicity,cytotoxity, | en |
| dc.relation.page | 129 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2015-02-16 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 食品科技研究所 | zh_TW |
| 顯示於系所單位： | 食品科技研究所 | |
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