請用此 Handle URI 來引用此文件：
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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 吳瑞碧(James Swi-Bea Wu) | |
| dc.contributor.author | Jian-Nan Chen | en |
| dc.contributor.author | 陳建男 | zh_TW |
| dc.date.accessioned | 2021-06-15T04:55:37Z | - |
| dc.date.available | 2015-08-04 | |
| dc.date.copyright | 2010-08-04 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-07-29 | |
| dc.identifier.citation | 參考文獻:
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46148 | - |
| dc.description.abstract | 菇類已成為受歡迎的抗腫瘤、抗病毒、抗菌、抗氧化和免疫調節劑的天然來源。Pleurotus citrinopileatusu別名玉皇菇為東方國家中常見的一種食用菇類,我們從其新鮮子實體分離出一種非凝集素的功能性醣蛋白(PCP-3A)並評估其免疫調節和抗腫瘤活性。電泳分析證實，PCP- 3A是一種由10個次單體所組成的醣蛋白，每個次單體大小約45 kDa。在體外培養細胞實驗發現，PCP- 3A能夠抑制U937增生且具有時間依存性。藉由流式細胞儀分析其抑制之機制，發現PCP-3A可藉由誘導U937細胞之S期停滯和細胞凋亡而抑制細胞增生。而在免疫調節活性方面， PCP-3A能刺激人類單核球細胞分泌細胞激素TNF-α、INF-γ、IL - 2和IL -10。此外，在抗發炎之活性作用上，PCP-3A能抑制經LPS誘導的RAW 264.7細胞之NO生成，分析PCP3 A對LPS誘導的RAW 264.7細胞中 iNOS和COX – 2之表現量，其中NO之減少和iNOS表現量之降低具有正相關性。 | zh_TW |
| dc.description.abstract | Mushrooms have become popular sources of natural antitumor, antiviral, antibacterial, antioxidative, and immunomodulatory agents. Golden oyster mushroom, Pleurotus citrinopileatus, is a common mushroom in oriental countries for human consumption. In this study, we isolated a non-lectin functional protein (PCP-3A) from the fresh fruiting body of this mushroom and investigated its immunomodulatory and antitumor activities. Electrophoresis demonstrated that PCP-3A is a glycoprotein composed of 10 subunits, each approximately 45.0 kDa in size. In vitro cell study showed that PCP-3A inhibits the proliferation of human tumor cell line U937 in a time-dependent manner. To elucidate the mechanism of growth inhibition, flow cytometry revealed that it is capable of inhibiting the growth of U937 cells by way of S phase arrest and apoptotic induction. In addition, evaluation of the immunomodulatory effect showed that PCP-3A stimulated the secretion of cytokines TNF-α, INF-γ, IL-2, and IL-10 from human peripheral blood mononuclear cells. The anti-inflammatory of PCP-3A was also evaluated. PCP-3A was found to effectively inhibit NO generation on LPS-Induced RAW 264.7 cells. To elucidate the mechanism of NO reduction, the effects of PCP3-A on LPS-induced iNOS and COX-2 expression in RAW 264.7 cells were examined.There is positive correlation between the inhibition on the generation of NO and the expression of inducible nitric oxide synthase in macrophage. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T04:55:37Z (GMT). No. of bitstreams: 1 ntu-99-D94641001-1.pdf: 1205801 bytes, checksum: 42ec06acf1fac5ca24701c3b1ed341da (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 目錄
頁碼 中文摘要.............................I 英文摘要.............................II 目錄...............................IV 圖次.............................. IX 表次..............................XI 第一章 前言............................. 1 第二章 文獻整理........................... 2 一、癌症.............................. 2 (一) 癌症簡介........................ 2 (二) 白血病.......................... 2 (三) 白血病細胞株....................... 4 二、免疫反應........................... 4 (一) 先天性免疫........................ 5 (二) 後天性免疫........................ 6 (三) 細胞激素......................... 7 三 食用菇.............................. 8 (一) 食藥用菇簡介....................... 8 (二) 菇類之膳食功能................................. 9 (1) 第一級功能：營養成分................... 9 (2) 第二級功能：嗜好特性................... 10 (3) 第三級功能：生理活性................... 10 (三) 菇類生理活性及相關研究.................. 11 (1) 抗腫瘤活性....................... 11 (2) 免疫調節活性...................... 13 (3) 降血壓作用....................... 14 (4) 降血脂作用.............................. 15 (5) 降血糖作用............................ 15 (6) 抗氧化作用............................. 16 (7) 抗病毒物質....................... 16 (8) 抗生素....................... 17 (9) 抗發炎作用...................... 17 (10) 抗菌作用......................... 18 (四)玉皇菇之介紹...................... 19 第三章 材料與方法.......................... 21 一. 材料與藥品....................... 21 1.原料........................................................ 21 2.細胞株........................................... 21 3.試驗藥品.......................................... 21 3-1.細胞培養用品......................... 22 4.細胞培養用品........................................ 23 5.儀器設備........................................................................ 23 6.套組............................................ 24 二. 菇類活性物質之純化及生理活性評估............................... 25 1.玉皇菇40%~80%飽和度硫酸銨沉澱物之製備........... 25 2.玉皇菇40%~80%飽和度硫酸銨沉澱物之離子交換樹脂......................... 25 3.玉皇菇離子交換樹脂區分物之細胞活性評估........................... 26 3-1.U937細胞株之冷凍、解凍與培養................ 26 3-2.菇類萃取物對U937細胞株之生長抑制測試........... 26 3-3.細胞計數及生長抑制率.................................... 26 4.蛋白濃度定量分析....................... 27 5.玉皇菇離子交換樹脂區分物之SDS-PAGE及Native-PAGE檢定法.. 27 5-1. SDS-PAGE電泳分析法.................... 27 5-2. Native-PAGE電泳分析法.................... 30 5-3.Coomassie Brilliant Blue R (CBR) 染色法.............................. 32 5-4.Periodic acid-Schiff (PAS) 醣蛋白染色法............................ 32 6.玉皇菇離子交換樹脂區分物之膠體管柱層析........................... 32 7.等電點分析......................................... 33 7-1.試劑............................. 33 7-2.電泳及染色方法........................ 33 8.PCP-3A之血液凝集反應................................................. 34 8-1. 4%紅血球懸浮液的製備.................. 34 8-2.血液凝集反應......................... 34 9. PCP-3A之胺基酸組成分析................................... 34 10. PCP-3A之N端定序分析..................................... 34 11PCP-3A之LC/MS/MS分析.................... 35 12.PCP-3A對U937細胞株之生長抑制測試............ 35 12-1.細胞與樣品培養...................... 35 12-2.細胞計數及生長抑制率................... 35 13.細胞週期分析....................... 35 14.PCP-3A對U937細胞株之間接生長抑制測試.......... 36 14-1.人類單核球細胞分離.................... 36 14-2.人類單核球細胞與樣品之培養................ 36 15單核球細胞條件培養液（MNC-CM）中細胞激素分析...... 37 15-1.試劑........................... 37 15-2.實驗方法......................... 37 16.PCP-3A對巨噬細胞RAW 264.7之抑制生長測試......... 38 17.PCP-3A對巨噬細胞RAW 264.7之NO 含量的測定........ 38 18.PCP-3A對巨噬細胞RAW 264.7之PGE2含量的測定....... 39 19.PCP-3A對巨噬細胞RAW 264.7之COX-2及iNOs蛋白質表現量定......................39 19-1.細胞萃取液收集.............................................. 39 19-2.電泳及轉漬........................ 40 20.統計分析.............................................. 40 第四章 結果與討論.......................... 41 1.玉皇菇40%~80%硫酸銨沉澱物之製備............................. 41 2.玉皇菇40%~80%硫酸銨沉澱物之離子交換樹脂區分....................... 41 3.玉皇菇離子交換樹脂區分物之細胞活性評估........................... 41 4.玉皇菇離子交換樹脂區分之SDS-PAGE及Native-PAGE分析... 42 5. 玉皇菇離子交換樹脂區分物之膠體管柱析........... 43 6. 玉皇菇膠體層析區分物之細胞活性評估............ 43 7. 活性蛋白PCP-3A之原態分子量分析及等電點分析....................... 44 8. 血液凝集反應分析..................... 44 9.PCP-3A之胺基酸組成分析.................. 45 10.PCP-3A之LC/MS/MS分析.................. 46 11.PCP-3A之N端定序分析................... 46 12.PCP-3A對U937細胞株之生長抑制測試............ 47 13.PCP-3A對U937細胞週期分布................ 47 14.PCP-3A對U937細胞株之間接生長抑制測試........................................ 49 15.單核細胞條件培養液（MNC-CM）中細胞激素分析........ 50 16.PCP-3A對巨噬細胞RAW 264.7之抑制............. 51 17.PCP-3A對巨噬細胞RAW 264.7之NO 含量及PGE2的測定.... 52 18.PCP-3A對巨噬細胞RAW 264.7之COX-2及iNOs蛋白質表現量測定...........................53 第五章總結............................ 54 參考文獻............................. 92 | |
| dc.language.iso | zh-TW | |
| dc.title | 玉皇菇中一種新免疫調節蛋白之純化與其生理活性之研究 | zh_TW |
| dc.title | Purification and Bioactivity of a Novel Immunomodulatory Protein from Pleurotus citrinopoleatus | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.coadvisor | 張鴻民(Hung-Min Chang) | |
| dc.contributor.oralexamcommittee | 何其儻,王進崑,吳明昌,江啟銘,廖慧芬,王裕泰,張雲祥 | |
| dc.subject.keyword | 抗癌,抗發炎,免疫調節,菇類,醣蛋白, | zh_TW |
| dc.subject.keyword | antitumor,anti-inflammatory,immunomodulatory,mushroom,glycoprotein, | en |
| dc.relation.page | 113 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2010-07-30 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 食品科技研究所 | zh_TW |
| 顯示於系所單位： | 食品科技研究所 | |
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