探讨灵芝子实体及深层培养菌丝体之多醣体结构特徵与其对免疫调节.doc

探討靈芝子實體及深層培養菌絲體之多醣體結構特徵 與其對免疫調節之影響 Comparisons of immunomodulatory effects and structural features of polysaccharides fractionated from fruiting body and submerged culture of Ganoderma lucidum 研究生：王韞皓 Wang, Yun-Hao 指導教授：郭嘉信 Guo, Jia-Shin 【摘要】 靈芝Ganoderma lucidum 萃取物具有調節免疫反應，增加人體的先天性及後天性免疫功能。靈芝萃取物之主要成分為β-多醣體β-polysaccharides 、三帖類triterpenes 、蛋白質、核酸、固醇steroids 等，而其中最受矚目且研究生理活性最多的應屬高分子多醣體，高分子多醣體會與免疫細胞作用而啟動免疫細胞內的訊息傳遞，進而活化免疫細胞，增強淋巴細胞的增生能力及加強巨噬細胞的吞噬能力，以達到增進人體免疫功能之效果。靈芝菌絲體培養液經酒精沉澱之多醣體與靈芝子實體經30℃或80℃水萃獲得之多醣體經FPLC搭配S300管柱分離純化後，獲得不同分子量多醣體之液相層析指紋圖譜及分液樣品fractionated samples ，實驗規畫，分別進行分子量、單醣組成及鍵結型態的鑑定，並測定其免疫細胞增殖能力、巨噬細胞吞噬能力及細胞激素的釋放量，以了解多醣體結構特徵與其免疫活性間的關聯性。結果發現，經純化後的多醣體分液樣品，分子量約分佈在1 – 190 kDa之間，且其化學結構是由β- 1→3 與β- 1→6 的葡萄糖組成佔多數，其餘還包含β- 1→4 等鍵結型態存在，而在細胞測試中發現，經純化後的多醣體分液樣品誘導總免疫細胞增殖及T細胞與B細胞增殖可達控制組的3.7 – 4.22倍；吞噬活性方面，可誘導巨噬細胞吞噬能力高於控制組21 %。由此可知，純化後的靈芝多醣具有增進免疫活性之功能，且經結構鑑定後判定，各分液樣品皆具同樣的化學結構組成。關鍵字：靈芝，多醣體，免疫調節 【Abstract】Ganoderma lucidum are considered to be capable of regulating immunomodulation and boosting innate and acquired immune systems in human body. The major components of the Ganoderma lucidum extracts are β-polysaccharides, triterpenes, proteins, nucleic acids, steroids, etc. Among those compounds, β-polysaccharides are the one which attract the most attention as well as research the most often on their physiological functions. β-Polysaccharides can interact with immunocytes and trigger their signal transductions, which may then activate the immunocytes, increase the multiplication of lymphocytes, and enhance the phagocytosis of macrophages, to boost up the functions of the human bodys immune system. we are using a FPLC equipped with a S300 column to fractionate the polysaccharides of Ganoderma lucidum from different sources to obtain polysaccharides fingerprintings and their fractionated samples containing polysaccharides of different molecular weight . In this research, The molecular weight, saccharide compositions and structure linkage of each fractionated polysaccharide were de