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| 題名: | 兩種真菌醱酵液之生物活性成分研究 |
| 作者: | 張雅雯 |
| 貢獻者: | 生藥學研究所 |
| 日期: | 2010 |
| 上傳時間: | 2010-10-21 09:11:31 (UTC+8) |
| 摘要: | 利用抑制一氧化氮合成酶的活性分析平台來篩選真菌培養株，藉以篩檢出具有抗發炎活性的真菌株，發現某些真菌株的醱酵培養液或菌絲體萃取物，對於RAW264.7細胞的產生一氧化氮具有顯著的抑制作用。遂選定：Theissenia rogersii（#92031201）及Stilbohypoxylon elaeicola（#173）兩株進行其活性成分研究。在選擇以麥芽抽取物（malt extract）為培養基加以擴大培養後，針對醱酵液所含代謝產物進行分析與分離，計獲得12個化合物，分別為： 2,5-dihydroxymethylfuran（1）、pycnidione（2）、maltol（3）、tyrosol（4）、hymatoxin K（5）、hymatoxin L（6）、elaeicolactone（7）、5-hydroxymethylfurfural（8）、elaeicolasdie A（9）elaeicolaside B（10）、elaeicolaside C（11）和 elaeicolaside D（12），其中屬於倍半萜之化合物7，及屬於二萜糖苷的化合物9、10及12，皆為新化合物。在100 μM的濃度下，10對於RAW264.7細胞的產生一氧化氮具有顯著的抑制活性且不具細胞毒性。此外，2及10對於A549肺腺癌細胞具有強的毒殺作用，其GI50分別為5.25 nM及9.79 μM。
The ethyl acetate extracts of the fermented broths of Theissenia rogersii (#92031201) and Stilbohypoxylon elaeicola (#173) were found to exhibit significant activity against inducible nitric oxide synthase (iNOS) in our preliminary screening. Therefore, bioassay-guided fraction and separation of the active component from these two broths were carried out which resulted in the isolation of twelve compounds. Their structures were elucidated to be 2,5-dihydroxymethylfuran (1), pycnidione (2), maltol (3), tyrosol (4), hymatoxin K (5), hymatoxin L (6), elaeicolactone (7), 5-hydroxymethylfurfural (8), elaeicolasdie A (9), elaeicolaside B (10), elaeicolaside C (11) and elaeicolaside D (12). Of these compounds identified, 7, 9, 10 and 12, classified as respective a sesquiterpene and three diterpene glycosides, were novel chemical entities. Compound 10 exhibited significant inhibitory activity on NO production of RAW264.7 cells without any cytotoxicity. In addition, 2 and 10 exerted potent cytotoxicity against an A549 lung cancer cell line with GI50 values of 5.25 nM and 9.79 μM, respectively. |
| 關聯: | 181頁 |
| 描述: | 摘要 I
Abstract II
總目錄 III
圖目錄 VII
縮寫表 XII
壹、緒論與研究目的 1
貳、炭角菌科真菌之天然物文獻回顧 5
参、實驗結果與討論 34
3.1.1 2,5-Dihydroxymethylfuran（1）之結構解析 39
3.1.2 Pycnidione（2）之結構解析 43
3.1.3 Maltol（3）之結構解析 54
3.1.4 Tyrosol（4）之結構解析 58
3.1.5 Hymatoxin K（5）之結構解析 61
3.1.6 Hymatoxin L（6）之結構解析 73
3.1.7 Elaeicolactone（7）之結構解析 82
3.1.8 5-Hydorxymethylfurfural（8）之結構解析 92
3.1.9 Elaeicolaside A（9）之結構解析 96
3.1.10 Elaeicolaside B（10）之結構解析 106
3.1.11 Elaeicolaside C（11）之結構解析 116
3.1.12 Elaeicolaside D（12）之結構解析 126
3.2 一氧化氮濃度及細胞毒性測定結果 135
3.3 A549細胞生長抑制測試結果 137
3.4 討論 138
肆、實驗部分 141
4.1 儀器設備與試劑 141
4.2 真菌材料 143
4.3 培養基配置 143
4.4.1 #92031201 培養液之成分分離與純化 145
4.4.2 #173培養液之成分分離與純化 148
4.5 單糖組成分析 152
4.6 單糖旋光度測定 153
4.7 一氧化氮（NO）濃度之測定：Griess reagent assay 154
4.8 細胞毒性測試（alamarBlue assay） 157
4.9 細胞生長測試（sulforhodamine B, SRB） 159
4.10 各成分之物理數據 161
參考文獻 165
附錄一 173
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