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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 盧虎生(Huu-Sheng Lur) | |
dc.contributor.author | Ching-Wen Chang | en |
dc.contributor.author | 張景雯 | zh_TW |
dc.date.accessioned | 2021-06-14T16:44:32Z | - |
dc.date.available | 2013-08-16 | |
dc.date.copyright | 2011-08-16 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-08-12 | |
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The structure of arnillarizin, a new p rotoilludane sesquiterpenoid aromatic ester from Armillaria mellea. Chinese Chemical Letters, 2, 732-741. Yang, J. S., Su, Y. L., & Wang, Y. L. (1991). Two novel protoilludane norsesquiterpenoid ester, armillasin and armillatin, from Armillaria mellea. Planta Medica, 57, 4782-4801. Yang, J. S., Su, Y. L., & Yu, D. Q. (1993). Carbon nuclear magnetic resonance spectra of some protoilludane sesquiter penoid aromatic esters from Armillaria mellea. Journal of Chinese Pharmaceutical Sciences, 2, 112-171. Yilmaz, N., Solmaz, M., Turlkekul, I., & Elmasras, M. (2006). Fatty acid composition in some wild edible mushrooms growing in the middle Black Sea region of Turkey. Food Chemistry, 99, 168-174. Zang, J. P., Sheng, Y., & Lian, B. (2004). Research Progress of Armillariella mellea. Studies of Trace Elements and Health, 21, 47-50. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40309 | - |
dc.description.abstract | 蜜環菌 (Armillariella mellea) 是中國傳統的中草藥,在過去的研究中被指出其內含多醣體成份具抗發炎能力。因此,得到兼具產量與品質的蜜環菌,便是本研究的目標。本研究發現在培養四十九天時,蜜環菌的乾重產量可以達到15.73 ± 0.16 g/l,同時多醣體和酒精萃取物的產量也可以分別達在到0.63 ± 0.04 g/l和5.51 ± 0.38 g/l。蜜環菌多醣體中以半乳糖和葡萄糖為主要成份,含量分別為1071.30 ± 10.89 和525.45 ± 4.14 mmol/g polysaccharide。蜜環菌酒精萃取物中含有成份ADP、cytidine和adenosine,在每毫克酒精萃取物中含量分別為0.48、0.34和0.74 mg。在抗發炎能力測定中,將蜜環菌萃取出來的多醣體、含硫多醣體和酒精萃取物分別處理在小鼠巨噬細胞RAW264.7及人類臍靜脈細胞EAhy926上,並分別以脂多醣體 (lipoplysaccharide) 和腫瘤壞死因子 (tumor necrosis factor-a) 刺激細胞使其發炎後,偵測tumor necrosis factor-a (TNF-a)、interleukin-6 (IL-6) 和monocyte chemotactic protein-1 (MCP-1) 的含量。結果發現:在RAW264.7中,酒精萃取物的濃度和TNF-a及IL-6的產量有劑量正相關。此外,培養四十九天的蜜環菌多醣體500 mg/ml顯著抑制TNF-a 及IL-6分泌的能力達31.17%和62.74%。含硫多醣體對TNF-a及IL-6產量的抑制達100%及56.82%,其效果優於不含硫的多醣體。EAhy中MCP-1的含量可被培養四十九天蜜環菌的多醣體、酒精萃取物和含硫多醣體抑制到52.17%、37.49%和34.53%。以上結果皆證實蜜環菌萃取物有抗發炎的效果。綜上所述,在本研究的培養條件下,蜜環菌乾重和多醣體的產量在培養四十九天後都能有效的被提升,而半乳糖和葡萄糖則是蜜環菌多醣體中含量最多的單糖成份。除此之外,無論是蜜環菌的多醣體、含硫多醣體或酒精萃取物,都能有效抑制TNF-a、IL-6 和MCP-1等和發炎相關的細胞激素。 | zh_TW |
dc.description.abstract | Armillariella mellea was reported to exhibit anti-inflammatory activity, especially that of the polysaccharide (PS) . To mass produce high quality of A. mellea was the aim of the study. The maximum production of 15.73 ± 0.16 g/l was obtained at 49 days, with the yield of PS and ethanolic extract of 0.63 ± 0.04 g/l and 5.51 ± 0.38 g/l, respectively. Galactose and glucose was the major sugar of PS with value of 1071.30 ± 10.89 and 525.45 ± 4.14 mmol/g PS at 49-day-culture, respectively. The components of ethanolic extract included ADP, cytidine and adenosine with the value of 0.48, 0.34 and 0.74 mg/mg ethanolic extract, respectively. Effects of extracted PS, sulfated polysaccharide (SPS) and ethanolic extract from cultured A. mellea on lipopolysaccharide (LPS) and TNF-a induced inflammation-related cytokines in RAW264.7 macrophages and EAhy926 were evaluated. Secretions of tumor necrosis factor-a (TNF-a) and interleukin-6 (IL-6) were detected in RAW264.7. The results showed that inhibition of TNF-a and IL-6 production were in a dose-dependent manner with ethanolic extract of A. mellea. PS from 49-day-cultured mycelia at 500 mg/ml showed significant inhibition of TNF-a and IL-6 in the value of 31.17% and 62.74%, respectively. Furthermore, SPS exhibited more effective on inhibition of TNF-a and IL-6 secretion as compared that with non-SPS in the value of 100% and 56.82% suppression, respectively. On the other hand, the secretion of cytokine monocyte chemotactic protein-1 (MCP-1) was significantly reduced in 52.17%, 37.49% and 34.53% for 49-day-culture PS, ethanolic extract and sulfated-PS (SPS), respectively. All of the above results suggested that A. mellea exhibited anti-inflammatory activity. In conclusion, in our cultured condition, A. mellea's dry weight and PS production could be enhanced at 49-cultured days. Glactose and glucose was the major component of A. mellea PS. PS, ethanolic extract and SPS of A. mellea significantly inhibited the release of the inflammatory related cytokines of TNF-a, IL-6 and MCP-1. | en |
dc.description.provenance | Made available in DSpace on 2021-06-14T16:44:32Z (GMT). No. of bitstreams: 1 ntu-100-R98621103-1.pdf: 2288682 bytes, checksum: ce739549ee656b50e5787e1edc5bc513 (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 誌謝 i
中文摘要 iii ABSTRACT iv CONTENTS v LIST OF FIGURES ix 第一章 緒論 1 第二章 前人研究 4 1. 蜜環菌 Armillariella mellea (Vahl.:Fr.) Karst. 4 1.1 生物型態 4 1.1.1 分類地位 4 1.1.2 形態與分布 5 1.2 成份 7 1.3 栽培特性 9 1.3.1 栽培方法 9 1.3.1.1 固體栽培技術 9 1.3.1.2 液體發酵培養 11 1.3.1.3 子實體培養 11 1.4 生物活性 12 1.4.1 抗發炎能力 12 1.4.2 抗腫瘤能力 13 1.4.3 抗血栓形成能力 13 1.4.4 對腦部缺血保護作用 14 1.4.5 抗血糖升高能力 14 1.4.6 細胞毒性測試 15 第三章 材料與方法 16 1.材料 16 1.1菌株 16 1.2 細胞 16 1.3 藥品及試劑 17 1.4 儀器及設備 19 2. 方法 22 2.1 培養基之配製方法 22 2.1.1 固態培養基 22 2.1.2 液態培養基 22 2.2 菌絲培養方法 23 2.3 生長曲線之測定 23 2.4 菌絲體萃取物之分離 24 2.5 多醣體分子量分佈分析 26 2.6 單醣組成分析 27 2.7 乙醇萃取物成份分析 28 2.8 細胞培養方法 29 2.9 SRB分析 30 2.10 抗血管新生能力測定 31 2.11 TNF-α 蛋白質ELISA 測定 32 2.12 IL-6 蛋白質ELISA測定 33 2.13 MCP-1蛋白質ELISA測定 34 2.14 西方墨點蛋白質分析 35 2.15 RNA萃取 36 2.16 PCR 37 2.17 統計分析 37 第四章 結果 38 1.生長曲線 38 2.多醣體產量及成份分析 41 2.1產量曲線 41 2.2多醣分子量分布 43 2.3單醣組成分析 45 3.乙醇萃取物產量及成份分析 48 3.1 產量曲線 48 3.2 成份分析 50 4. 甲醇萃取物產量 53 5. 多醣體細胞毒殺能力分析 55 6. 抗血管新生能力測試 57 7. 多醣體及乙醇萃取物對RAW264.7細胞製造TNF-α的影響 59 8. 多醣體及乙醇萃取物對RAW264.7細胞製造IL-6的影響 66 9. 多醣體及乙醇萃取物對EAhy926細胞製造MCP-1的影響 72 10. 蜜環菌多醣體、含硫多醣體及乙醇萃取物對IkB-α表現之影響 78 11. 蜜環菌多醣體及乙醇萃取物對TNF-α及IL-6基因表現之影響 81 第五章 討論 84 1.蜜環菌及其多醣體、乙醇萃取物、甲醇萃取物之產量變化 84 2 .蜜環菌多醣體定性分析 86 3.蜜環菌乙醇萃取物成份討論 88 4 .蜜環菌多醣體及乙醇萃取物、含硫萃取物之抗發炎能力比較 89 第六章 結論 91 第七章 參考文獻 92 LIST OF FIGURES 圖 1蜜環菌菌絲體萃取流程 25 圖 2蜜環菌培養49天之外表型 39 圖 3蜜環菌生長曲線 40 圖 4蜜環菌多醣體產量曲線 42 圖 5不同培養天數之蜜環菌多醣體分子量分布變化 44 圖 6比較不同培養天數的蜜環菌多醣體之單醣含量變化 46 圖 7蜜環菌乙醇萃取物產量曲線 49 圖 8不同培養天數之蜜環菌乙醇萃取物成份的含量變化 51 圖 9蜜環菌甲醇萃取物產量曲線 54 圖 10蜜環菌多醣體抗血管新生能力測試 58 圖 11不同蜜環菌多醣體濃度對LPS誘導RAW264.7製造TNF-α的影響 61 圖 12不同蜜環菌乙醇萃取物濃度對LPS誘導RAW264.7製造TNF-α的影響 62 圖 13不同培養天數蜜環菌萃取之多醣體對LPS誘導RAW264.7細胞TNF-α製造之變化 63 圖 14不同培養天數蜜環菌萃取之乙醇萃取物對LPS所誘導RAW264.7細胞TNF-α製造之變化 64 圖 15培養49天之蜜環菌多醣體及含硫萃取法所得之多醣體,對LPS所誘導RAW264.7細胞TNF-α製造之變化 65 圖 16不同蜜環菌多醣體濃度對LPS誘導RAW264.7製造IL-6的影響 67 圖 17不同蜜環菌乙醇萃取物濃度對LPS誘導RAW264.7製造IL-6的影響 68 圖 18不同培養天數蜜環菌萃取之多醣體對LPS誘導RAW264.7細胞IL-6製造之變化 69 圖 19不同培養天數蜜環菌萃取之乙醇萃取物對LPS誘導RAW264.7細胞IL-6製造之變化 70 圖 20培養49天之蜜環菌多醣體及含硫萃取法所得之多醣體,對LPS所誘導RAW264.7細胞IL-6製造之變化 71 圖 21不同蜜環菌多醣體濃度對TNF-α誘導EAhy926製造MCP-1之影響 73 圖 22不同蜜環菌乙醇萃取物濃度對TNF-α誘導EAhy926製造MCP-1之影響 74 圖 23不同培養天數蜜環菌萃取之多醣體對TNF-α誘導EAhy926細胞MCP-1製造之變化 75 圖 24不同培養天數蜜環菌萃取之乙醇萃取物對TNF-α所誘導EAhy926細胞MCP-1製造之變化 76 圖 25培養49天之蜜環菌多醣體及含硫萃取法所得之多醣體,對TNF-α所誘導EAhy926細胞MCP-1製造之變化 77 圖 26培養49天之蜜環菌多醣體、含硫多醣體和酒精萃取物,對LPS誘導RAW264.7 細胞內IkB-a降解之影響 79 圖 27培養49天之蜜環菌多醣體、含硫多醣體和酒精萃取物,對TNF-α誘導EAhy926 細胞內IkB-a降解之影響 80 圖 28 培養49天之蜜環菌多醣體和酒精萃取物,對LPS誘導之TNF-α基因在RAW 264.7細胞中表現之影響 82 圖29培養49天之蜜環菌多醣體和酒精萃取物,對LPS誘導之IL-6基因在RAW264.7細胞中表現之影響 83 LIST OF TABLES 表 1 蜜環菌的成份 7 表2 Pullulan之組成 26 表 3 HPLC之流洗條件 28 表 4 SRB分析使用之細胞株培養條件 30 表 6. 蜜環菌培養7至49天所萃取得到的乙醇萃取物內含的不同成份在培養期間內的含量變化 52 表 7.蜜環菌多醣體細胞毒殺能力測試結果 56 | |
dc.language.iso | zh-TW | |
dc.title | 蜜環菌栽培體之成份分析與抗發炎活性研究 | zh_TW |
dc.title | Studies on chemical constituents and anti-inflammatory activity of cultivated Armillariella mellea | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 盧美光(Mei-Kuang Lu) | |
dc.contributor.oralexamcommittee | 鄭靜枝(Jing-Jy Cheng),劉啟德(Chi-Te Liu) | |
dc.subject.keyword | 蜜環菌,發炎,細胞激素, | zh_TW |
dc.subject.keyword | Armillariella mellea,anti-inflammatory,TNF-a,IL-6,MCP-1,IkB-a, | en |
dc.relation.page | 101 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-08-12 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 農藝學研究所 | zh_TW |
顯示於系所單位: | 農藝學系 |
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