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標題: | 牛樟芝多醣體之免疫活性分析及結構鑑定 Structural determination of immunoactive polysaccharides from Antrodia cinnamomea |
作者: | Shih-Hao Zhan 詹世豪 |
指導教授: | 吳世雄(Shih-Hsiung Wu) |
關鍵字: | 牛樟芝,多醣,免疫活性,膠體過濾,氣相層析質譜儀,腫瘤壞死因子,一氧化氮, Antrodia cinnamomea,polysaccharides,immunoactivity,gel filtraton,GC-MS,TNF,NO, |
出版年 : | 2010 |
學位: | 碩士 |
摘要: | 牛樟芝傳統上多用來做為中藥用,先前的研究也指出,牛樟芝多醣體擁有免疫調節上的活性,而對於它的多醣的結構鑑定尚未明瞭。
首先利用氰化甲烷以及冷熱水來萃取牛樟芝菌絲體,之後將水層所得到的多醣再經由膠體過濾層析法來分離,得到了三個不同分子量大小的多醣體,依序為 HMPS、LT70和GT70。根據生物活性以及多醣產量的結果,我們選擇了LT70並利用離子交換樹脂層析法來做進一步的純化萃取並且獲得了三部分的多醣樣本(QA、QB和QC)。 在本篇研究中,牛樟芝多醣刺激了一氧化氮(NO)產生,並且也進一步測試了牛樟芝參與老鼠巨噬細胞在腫瘤壞死因子-α(TNF-α)以及細胞介質素-6(IL-6)的產生。由氣相層析質譜儀的分析結果上,QB的主要醣類成分組成為甘露糖和半乳糖且兩者比例約為4:1。同時也利用氣相層析質譜儀分析在QB樣本中含有:鍵結terminal mannopyranose,1,2 linked galactopyranose,1,3 linked mannopyranose,以及 1,2,6 linked mannopyrnose。同時並利用化學以及光譜分析來鑑定由牛樟芝所分離出的活性多醣QB結構,初步推測應為: 而單醣D應是接在A或C的6號位置的一個分支醣基。然而完整的結構待需進一步的分析來鑑定。 Antrodia cinnamomea is used as traditional Chinese medicine. In previous studies, polysaccharides from A. cinnamomea possess immune-modulating activity, and the structure determination is still not clear. Mycelia of A. cinnamomea were first extracted with acetonitrile and followed with hot and cold water, in which the extract was further separated by gel filtration chromatography to give three fractions containing polysaccharides HMPS, LT70, and GT70. Due to the biological activities and amounts of polysaccharides, LT70 was selected for further purification and identification. By using ion exchange chromatography, LT70 was separated into three polysaccharide fractions; QA, QB, and QC. In the present study, A. cinnamomea polysaccharides participated in the stimulation of nitric oxide(NO) production and stimulation of murine wild-type macrophages predominantly in tumor necrosis factor-α (TNF-α) and interleukin-6(IL-6) production. From sugar analysis by gas-chromatography mass spectrum(GC-MS), the major components of A. cinnamomea polysaccharides QB are mannose and galactose in the molar ratio of 4:1. The main glycosidic linkages were also characterized by GC-MS. The results showed that there are terminal mannopyranose, 1,2 linked galactopyranose, 1,3 linked mannopyranose, and 1,2,6 linked mannopyranose. The structures of bioactive polysaccharides from A. cinnamomea were determined by chemical and NMR methods, and we preliminarily presume that the structure is as following: And D might be a 6 linked monosaccharide to A or C as a branched terminal residue.The completed structure must be determined by further analysis. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/27718 |
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顯示於系所單位: | 生化科學研究所 |
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