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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 許 輔 | |
dc.contributor.author | Jeng-Je Yang | en |
dc.contributor.author | 楊政哲 | zh_TW |
dc.date.accessioned | 2021-06-14T16:56:48Z | - |
dc.date.available | 2009-08-05 | |
dc.date.copyright | 2008-08-05 | |
dc.date.issued | 2008 | |
dc.date.submitted | 2008-07-29 | |
dc.identifier.citation | 有地滋。1985。靈芝的奇妙功效。青春出版社。台北市。
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Study of Ganoderma lucidum polysaccharides on effects of cellular immune function in mice. J. Microbiol. 23: 51-54. Kino K., K. Mizumoto, T. Sone, T. Yamaji, J. Watanabe, A. Yamashita, K. Yamaoka, k. Shimizu, and K. Ko. 1990. An immunomodulating protein, Ling Zhi-8 (LZ-8) prevents insulitis in non-obese diabetic mice. Diabetologia 33: 713-718. Kino, K., T. Sone, J. Watanabe, A. Yamashita, H. Tsuboi, H. Miyajima, and H. Tsunoo. 1991. Immunomodulator, LZ-8, prevents antibody production in mice. Int. J. Immunopharmac. 13: 1109-1115. Kino, K., A. Yamashita, K. Yamaoka, J. Watanabe, S. Tanaka, K. Ko, K. Shimizu, and H. Tsunoo. 1989. Isolation and characterization of a new immunomodulatory protein, Ling Zhi-8 (LZ-8), from Ganoderma lucidum . J Biol Chem. 264: 472-478. Kohguchi, M., T. Kunikata, H. Watanabe, N. Kudo, T. Shibuya, T. Ishihara, K. Iwaki, M. Ikeda, S. Fukuda, and M. Kurimoto. 2004. 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Potential of a novel polysaccharides preparation (GLPP) from anhui-grown Ganoderma lucidum in tumor treatment and immunostimulation. 72: 435-442. Sliva, D. 2004. Cellular and physiological effects of Ganoderma lucidum (Reishi). Mini Rev Med Chem. 4: 873-879. Shao, B. M., H. Dai, W. Xu, Z. B. Lin, and X. M. Gao. 2004. Immune receptors for polysaccharides from Ganoderma lucidum. Biochem Biophys Res Commun. 323: 133-141. Tanaka, S., K. Ko, K. Kino, K. Tsuchiya, A. Yamashita, A. Murasugi, S. Sakuma, and H. Tsunoo. 1989. Complete amino acid sequence of an immunomodulatory protein, Ling Zhi-8 (LZ-8). J Biol Chem. 264: 16372-16377. Tang, Q. J., J. S. Zhang, Y. J. Pan, R. Werner, and H. Fan. 2004. Activation of mouse macrophages by the alkali-extrated polysaccharide from spore of Ganoderma lucidum. Chin. J. Cell Mol. Immunol. 20: 142-144. Wang, S. Y., M. L. Hsu, H. C. Hsu, C. H. Tzeng, S. S. Lee, M. S. Shiao, and C. K. Ho. 1997. The anti-tumor effect of Ganoderma lucidum is mediated by cytokines released from activated macrophages and T lymphocytes. Int. J. Cancer: 609-705. Wang, Y. Y., K. H. Khoo, S. T. Chen, C. C. Lin, C. H. Wong, and C. H. Lin. 2001. Studies on the immuno-modulating and antitumor activities of Ganoderma lucidum (Reishi) polysaccharides: functional and proteomic analyses of a fucose-containing glycoprotein fraction responsible for the activities. Bioorg Med Chem. 10: 1057-1062. Xia, D., Z. B. Lin, R. Z. Li, and Y. Q. He. 1989. Effects of Ganoderma polysaccharides on immune function in mice. J. Beijing Med. Univ. 23: 329-333. Zhang, Q. H. and Z. B. Lin. 1999. Effects of Ganoderma lucidum polysaccharides B on TNF-alpha and IFN-gamma production and their mRNA expression. J. Beijing Med. Univ. 31: 179-183. Zhang, J., Q. Tang, Z. K. Martin, W. Reutter, and H. Fan. 2002. Activation of B lymphocytes by GLIS, a bioactive proteoglycan from Ganoderma lucidum . Life Sci. 71: 623-638. Zhu, X. L., A. F. Chen, and Z. B. Lin. 2006. Ganoderma lucidum polysaccharides enhance the function of immunological effector cells in immunosuppressed mice. J Ethnopharmacol. 111: 219-226. Zhu, X. L. and Z. B. Lin. 2005. Effect of Ganoderma lucidum polysaccharides on proliferation and cytotoxity of cytokine-induced killer cells. Acta Pharmacol. Sin. 26: 1130-1137. Zhu, X. L. and Z. B. Lin. 2005. Modulation of cytokines production, granzyme B and perforin in murine CIK cells by Ganoderma lucidum polysaccharides. Carb. Pol. 63: 188-197. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40707 | - |
dc.description.abstract | 靈芝 (Ganoderma lucidum (Fr.) Karst) 已知的功效成分包含多醣與免疫調節蛋白 LZ8,但是此兩者調節免疫之對象與機制之差異仍不清楚。本研究第一部份即為探討靈芝多醣PSG與蛋白LZ8對於小鼠腹腔巨噬細胞及脾細胞的活化作用,並檢視兩者對於調節細胞免疫功能之差異;第二部份則建立LZ8酵素連結免疫分析之定量平台,同時定量市售樣品中靈芝蛋白LZ8的含量,以及比較樣品中活性成分多寡與小鼠脾細胞活化作用之相關性。
第一部份結果發現,靈芝多醣PSG可活化小鼠腹腔巨噬細胞分泌TNF-alpha,但靈芝蛋白LZ8則是活化腹腔巨噬細胞分泌IL-12,顯示PSG與LZ8活化巨噬細胞分泌不同的細胞激素。另外,LZ8能直接活化小鼠脾細胞與 CD4+ T細胞產生IFN-gamma 與IL-2,同時提高 Th1 相關之細胞激素 mRNA 表現量,但PSG只能活化脾細胞分泌IL-2,且對於CD4+ T細胞則無活化的作用。另一方面,以PSG和LZ8分別活化小鼠腹腔巨噬細胞,再將活化後的巨噬細胞與 CD4+ T細胞共同培養,結果發現經過LZ8處理後的腹腔巨噬細胞,能促進CD4+ T細胞分泌IFN-gamma與IL-2,而PSG處理後之腹腔巨噬細胞,只能促進CD4+ T細胞分泌IL-2,且分泌量較少。此外, PSG與LZ8對TLR4缺失小鼠腹腔巨噬細胞共同培養後,PSG無法活化其分泌TNF-alpha,但仍可促進IL-1 beta 與IL-12的分泌,且PSG與TLR4缺失之腹腔巨噬細胞共同培養後,仍可促進CD4+ T分泌IL-2,故推測其活化的作用來自PSG中所含的微量LZ8。上述結果說明了靈芝蛋白LZ8對於活化T淋巴細胞的效果較顯著。 第二部份利用單株抗體建立 LZ8 之 ELISA 分析方法,其定量曲線(y = 0.0019 x + 0.2973)之線性係數 r2 = 0.9806,定量濃度範圍在0.15 – 200 μg/mL,以此方法來定量四種市售樣品中LZ8含量,其含量介於 0.58 - 11.76 μg/g。此外,分析各市售樣品中LZ8與PSG含量對活化脾細胞之相關性,發現樣品中LZ8含量多寡對於小鼠脾細胞IFN-gamma產生量,具有良好的相關性(r2 = 0.8502),因此 LZ8 含量與其活化小鼠脾細胞產生IFN-gamma之作用,可作為快速評估靈芝樣品活性成份與規格標準的指標。 | zh_TW |
dc.description.provenance | Made available in DSpace on 2021-06-14T16:56:48Z (GMT). No. of bitstreams: 1 ntu-97-R95628213-1.pdf: 834085 bytes, checksum: 00c452ecdf57f54e52cf303b30f48dd1 (MD5) Previous issue date: 2008 | en |
dc.description.tableofcontents | 摘要………………………………………………………………………………………I
Abstract…………………………………………………………………………...….…III 總目錄…………………………………………………………………………………..V 圖表目錄……………………………………………………………………………..VIII 壹、前人研究…………………………………………………………………………….1 一、 靈芝簡介…………………………………………………………………….1 二、 靈芝之免疫調節活性……………………………………………………….2 三、 靈芝多醣……………………………………………….……………………5 四、 靈芝免疫調節蛋白LZ8……………………………………………………10 五、 T淋巴細胞之分化作用……………………………………………………12 六、 研究動機與目的………………………………………..…………………..13 貳、材料與方法………………………………………………………………………..15 一、靈芝樣本對小鼠巨噬細胞之影響……………………………………………..15 (一) C57BL/6J小鼠腹腔巨噬細胞之取得…………….…………………….15 (二) 細胞激素TNF-alpha之測定………………………………………………….15 (三) 細胞激素IL-12之測定…………………………………………………17 (四) 細胞激素IL-1 beta之測定………………………..…………………………18 (五) 即時定量聚合酶連鎖反應…………………………………….………..19 二、靈芝樣本對小鼠脾臟細胞之影響……………………………………………..21 (一) C57BL/6J小鼠脾臟細胞之取得………………………………….…….21 (二) 小鼠T淋巴細胞之純化……………………………………….………….23 (三) 淋巴細胞增生試驗………………………………………………..……..24 (四) 細胞激素IFN-gamma之測定…………………………………………………..25 (五) 細胞激素IL-2之測定……………………………………………………26 (六) 即時定量之聚合酶連鎖反應…………………………………………….27 (七) 靈芝樣本刺激後之腹腔巨噬細胞與T細胞共同培養………………….30 三、建立 LZ8 的 ELISA 免疫分析平台…………………………………………31 (一) 蛋白質濃度測定…………………………………………………...……..31 (二) 總醣含量測定 (酚-硫酸法Phenol-Sulfuric Acid Method)……...………32 (三) LZ8 抗體生物素化 (biotinylation)…………………………..…………33 (四) LZ8 捕捉及偵測抗體配對之ELISA分析………………..……………34 (五) LZ8 濃度標準曲線建立…………………………………..…………….35 (六) 利用 LZ8 之 ELISA 定量平台分析靈芝樣品……………..…………36 参、結果………………………………………………………………………………..38 一、 靈芝多醣proteoglycan與去蛋白靈芝多醣PSG之特性分析………………38 (一) 利用蛋白酵素處理靈芝多醣以得到去蛋白靈芝多醣…………………….38 (二) 靈芝多醣與去蛋白靈芝多醣蛋白含量、總醣含量以及靈芝蛋白LZ8含量分析………………………………………………………………………...39 二、 PSG與LZ8對小鼠腹腔巨噬細胞的影響………………………………….…40 (一) PSG與LZ8分別活化小鼠腹腔巨噬細胞產生不同細胞激素…………….40 三、 PSG與LZ8對小鼠脾細胞的影響…………………………………………….42 (一) LZ8能直接活化小鼠脾細胞與CD4+ T細胞分泌IFN-gamma與IL-2,但PSG無此效果……………………………………………………………………….42 (二) 與LZ8共同培養後能提升CD4+ T細胞之Th1細胞激素 mRNA量的表 現…………………………………………………………………………….44 四、受LZ8活化之腹腔巨噬細胞能活化CD4+ T細胞產生IFN-gamma與IL-2,但PSG無此效果…………………………………………………………………...45 五、靈芝多醣PSG對於TLR4缺失小鼠 (C57BL/10ScN) 之腹腔巨噬細胞與脾細胞仍具活化作用………………………………………………………………….46 (一) PSG無法活化TLR4缺失小鼠之腹腔巨噬細胞產生TNF-alpha,但仍可活化IL-1 beta與IL-12的產生 (二) PSG與LZ8活化後之腹腔巨噬細胞促進T細胞產生IFN-gamma與IL-2之效果,在正常小鼠與TLR4缺失小鼠中有相同的趨勢……….………………….48 六、建立靈芝免疫調節蛋白之ELISA分析方法……………….………………...49 (一) LZ8捕捉及偵測抗體配對之ELISA分析…………………………………49 (二) LZ8濃度標準曲線建立 …………………………………………………....50 七、建立靈芝產品活性成分的規格標準……………………………………51 (一) 定量市售靈芝健康食品中LZ8 的含量…………………………......51 (二) 產品中活性成分多寡與活化小鼠脾細胞活性的相關性…………..51 肆、討論……………………………………………………………………………….54 伍、結論與未來展望………………………………………………………………….67 參考文獻……………………………………………………………………………….68 | |
dc.language.iso | zh-TW | |
dc.title | 研究靈芝多醣PSG與蛋白LZ8之免疫調節功效及成分規格標準 | zh_TW |
dc.title | Study on the Diverse Immunomodulatory Effects of Genoderma lucidum Polysaccharide PSG and Protein LZ-8 as well as the Establishment of Quality and Functional Standards for Genoderma Related Products | en |
dc.type | Thesis | |
dc.date.schoolyear | 96-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 許先業,柯俊良,繆希椿,謝光煬 | |
dc.subject.keyword | 靈芝多醣PSG,靈芝蛋白LZ8,免疫調節,免疫分析, | zh_TW |
dc.subject.keyword | Polysaccharide of Ganoderma lucidum (PSG),Ganoderma lucidum immunomodulatory protein LZ8,Immunomodulation,ELISA Immunoassay, | en |
dc.relation.page | 90 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2008-07-30 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 園藝學研究所 | zh_TW |
顯示於系所單位: | 園藝暨景觀學系 |
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