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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳水田(Shui-Tein Chen) | |
| dc.contributor.author | Chia-Che Tsai | en |
| dc.contributor.author | 蔡佳哲 | zh_TW |
| dc.date.accessioned | 2021-06-15T02:44:25Z | - |
| dc.date.available | 2010-08-14 | |
| dc.date.copyright | 2009-08-14 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-08-10 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/44196 | - |
| dc.description.abstract | 人類利用天然物作為預防以及治療疾病已有長久的歷史，本實驗利用單子葉植物小麥草以及靈芝多醣作為材料，鑑定其中具有免疫調控活性物質的結構與活性機制。利用細胞體篩選模式，我們發現小麥草的活性多醣成分WG-PS3 具有活化成人週邊血單核球細胞當中單核細胞、自然殺手細胞、T細胞之效果。WG-PS3直接活化單核細胞，提高CD69, CD80, CD86, IL-12, 以及TNF-a的表現。而WG-PS3 則利用直接活化單核細胞來間接影響NK細胞以及T細胞的活化。WG-PS3主要的結構型式為(a,b)-1,4-hexose, 利用HPLC的分離, 其中hepta-1,4-glucan 展現了顯著的免疫活化特性以及單核細胞的活化能力。
靈芝多醣F3是一個分子量大約500KD的巨分子，藉由細胞體篩選模式我們了解F3能直接活化成人週邊血單核球細胞當中單核細胞、自然殺手細胞、以及T細胞。利用史密斯降解法我們成功降解F3並且進一步分離降解產物。這些分離收集的降解產物都保留了活化單核細胞、自然殺手細胞、以及T細胞提高其CD69表現的能力，而其中一個部分F3S-G3擁有獨特的結構特徵 b-(1,4)-Glucorounic acid / Glucose oligomer 在NK細胞以及T細胞中表現顯著的IL-2細胞激素誘導能力。此外，F3S-G4也是值得注意的部分。F3S-G4的分子量大約是1000Dalton, 在細胞體篩選模式下發現其保留F3活化單核細胞、自然殺手細胞、T細胞之活性效果。F3S-G4可以成功的利用HPLC進行分離純化，可作為進一步鑑定特定完整結構的良好材料。 | zh_TW |
| dc.description.abstract | Natural products have played an important role throughout the world in preventing and treating human diseases throughout the history in different parts of the world. The Monocotyledoneae plant wheat grass and the Ganoderma lucidum polysaccharides were served as materials in this study to identify the molecules with immuno-modulating properties. By cytomic screening, the immuno-modulating behavior of the bioactive oligosaccharides (WG-PS3) from wheat grass has been described. WG-PS3 activated monocytes, T cells, and NK cells in human peripheral blood mononuclear cell (hPB-MNC). WG-PS3 was observed directly activate the purified monocytes by inducing the expression of CD69, CD80, CD86, IL-12, and TNF-a. In contrast, WG-PS3 indirectly affected NK and T cells only in the presence of monocytes. The structure of WG-PS3 was characterized suggesting repeating (a,b)-1,4-hexose. After HPLC separation, a hepta-1,4-glucan were identified from WG-PS3 and exhibited significantly ability to induce monocytes activation.
Ganoderma lucidum polysaccharides F3 were macromolecules with molecular weight around 500KD. F3 was found that processed the ability to directly induce NK cells, T cell, and monocytes activation by cytomic screening. By Smith degradation, we successfully degraded and fractionalized the F3. All of these fractions retained the CD69 induction activity to monocytes, T cells, and NK cells in hPB-MNC. However, a fraction F3S-G3 with structural signature of b-(1,4)-Glucorounic acid / Glucose oligomer was presented the specific immune property of low monocyte induction activity and high ability to stimulate IL-2 secretion in NK and T cells. In addition, the fraction F3S-G4 with low molecular weight around 1000 Dalton retained all the immuno-activating characteristics of F3 that described in above cytomic screening. The F3S-G4 involved heteroglycan and could be further separated by HPLC. Thus, the F3S-G4 may serve the materials for further analysis of defined structure. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T02:44:25Z (GMT). No. of bitstreams: 1 ntu-98-D93442006-1.pdf: 8361488 bytes, checksum: f5cb1f78cfd338b7a18fe46a9ac7355b (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | Table and Contents
謝 誌 ii 中 文 摘 要 iv Abstract v Abbreviations viii List of Figures ix List of Tables xii 1. Introduction 1.1 Natural products and immune modulation …………………………………..1 1.2 Wheat grass ………………………………………………………………2 1.3 Ganoderma Lucidum ………………………………………………………...3 1.4 The immune system and immune modulation………………………………..6 1.5 Cytomic screening platform for immuno-modulating nature products ………8 2. Materials and Methods 2.1 Chemicals and Antibodies …………………………………………………..11 2.2 Extraction and purification of immuno-modulating molecules from wheat grass ……………………………………………………………………….11 2.3 Smith degradation of Ganoderma Lucidum polysaccharides F3…………….12 2.4 Isolation and Stimulation of Adult Human Peripheral Blood Mononuclear Cells (hPB-MNCs) ………………………………………………………......13 2.5 Isolation of Monocytes ,T Cells, and NK cells from hPB-MNCs………...…14 2.6 Detections of cell surface CD Marker Expression by Flow Cytometric Assay…………………………………………………………………………15 2.7 Determination of immune cells activation by Q-PCR assay ………………...16 2.8 Determination of Th1 cytokine concentrations in medium by ELISA………17 2.9 Analyze the sugar compositions and linkages by GC-MS…………………...18 2.10 NMR ………………………………………………………………………19 2.11 2-AB (2-aminobenzamide) labeling of oligosaccharides………………….19 2.12 Separation of 2-AB labeling of oligosaccharides by normal phase HPLC..19 3. Results 3.1 Design the cytomic screening strategy to identify immuno-modulating molecules from nature plants ………………………………………………..21 Part1 : 3.1-1 The immunological active property of wheat grass extracts………………23 3.1-2 Cytomic screening and the immuno-activating mechanism of WG-PS3….24 3.1-3 Structural characterization of WG-PS3…………………………………....27 3.1-4 Structural identification and immuno-modulating activity examination of WG-PS3 derived fractions after HPLC……………………………………28 Part2 : 3.2-1 Cytomic screening and the immuno-activating mechanism of Ganoderma lucidum polysaccharides F3………………………………………………31 3.2-2 Smith degradation of F3 and cytomic screening of the fractionized smith degradation products……………………………………………………...33 3.2-3 Structural characterization of F3S-G3…………………………………….35 3.2-4 F3S-G4, a smith degradation fraction with low molecular weight exhibited immuno-modulaitng activities…………………………………………….35 4. Discussions…………………………………………..………………………..37 Reference List…………..………………………………………………………..41 Figure and Legend ………..……………………………………………………47 Research publications …………………………………………………………90 | |
| dc.language.iso | en | |
| dc.subject | 小麥草 | zh_TW |
| dc.subject | 單核球細胞 | zh_TW |
| dc.subject | 免疫調控 | zh_TW |
| dc.subject | 寡醣 | zh_TW |
| dc.subject | 靈芝多醣 | zh_TW |
| dc.subject | mononuclear cells | en |
| dc.subject | Ganoderma lucidum polysaccharides | en |
| dc.subject | oligosaccharides | en |
| dc.subject | immuno-modulation | en |
| dc.subject | wheat grass | en |
| dc.title | 單子葉植物小麥草與靈芝多醣之免疫活性分子結構
與功能機制探討 | zh_TW |
| dc.title | Structure and biological function of
immuno-modulating molecules of wheat grass and Ganoderma lucidum polysaccharides | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 吳世雄(Shih-Hsiung Wu),羅禮強(Lee-Chiang Lo),施子弼(Tzi-Bi Shih),曾銘仁(Min - Jen Tseng) | |
| dc.subject.keyword | 小麥草,靈芝多醣,寡醣,免疫調控,單核球細胞, | zh_TW |
| dc.subject.keyword | wheat grass,Ganoderma lucidum polysaccharides,oligosaccharides,immuno-modulation,mononuclear cells, | en |
| dc.relation.page | 90 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2009-08-10 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科學研究所 | zh_TW |
| 顯示於系所單位： | 生化科學研究所 | |
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