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研究生:何欣欣
研究生(外文):Xin-Xin He
論文名稱:鹿茸胜肽成分研究及iridoids免疫活性研究
論文名稱(外文):Study on the protein of Cervus elaphus L. antler and the immunology assay of iridoids.
指導教授:石麗仙
指導教授(外文):Li-Shian Shi
學位類別:碩士
校院名稱:國立虎尾科技大學
系所名稱:光電與材料科技研究所
學門:工程學門
學類:電資工程學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:64
中文關鍵詞:鹿茸環烯醚萜介白素-8介白素-10轉化生長因子-β
外文關鍵詞:Cervus elaphusiridoidIL-8IL-10TGF-β
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鹿茸為中國傳統名貴中藥,臨床實驗顯示鹿茸具有抗發炎、促進機體生長發育和新陳代謝、增強機體的免疫功能、抗氧化、抗老化、增強性功能、提高耐力、促進體內蛋白質合成、抗胃潰瘍、抗腫瘤及促進創傷癒合等功能。
由於市面上所販售之鹿茸產品良莠不一,因此本論文由市售馬鹿鹿茸切片中萃取得到鹿茸粗萃物(CEEL-P),並以此鹿茸粗萃物與市售的鹿茸產品(A、B、C廠商)進行胜肽含量分析及鹿茸胜肽成份分析比較,藉以分辨市售鹿茸產品之真偽優劣。實驗結果顯示,三種市售鹿茸產品中,其鹿茸蛋白質含量約為1 × 103 – 4.6 × 103 µg/ml,其中A廠商之鹿茸產品蛋白質含量最低,B廠商次之,C廠商最佳。由SDS-PAGE電泳圖顯示鹿茸多肽之含量與種類也以C廠商之鹿茸產品最佳。
Iridoid是常見的植物二次代謝產物,文獻指出iridoid及其苷類具有多種生物活性,如抗菌、抗氧化、抗發炎、鎮痛、保肝等。本論文針對12種iridoids化合物:stilbericoside (1)、6-epi-stilbericoside (2)、holmioside (3)、thunaloside (4)、2-O-primeverosylpaeonol (5)、6α-hydroxygeniposide (6)、gardenoside (7)、ixoroside (8)、geniposide (9)、shanzhiside (10)、genipin gentiobioside (11)、jasminoside B (12)進行免疫活性分析。於細胞存活率試驗中,化合物1-12於濃度1~10 µM下,對HT-29細胞株均無顯著的細胞毒性。免疫活性試驗結果顯示化合物1-5對HT-29細胞株有促進IL-8分泌的活性;化合物6-12則會抑制HT-29細胞株分泌IL-8,其中化合物7、10-12具有顯著的抑制HT-29細胞株分泌IL-8的活性。進一步以化合物7、10-12對HT-29細胞株分泌IL-10及TGF-β的含量變化研究。結果顯示化合物7、10-12不會刺激HT-29細胞株分泌IL-10;並具有顯著促進HT-29細胞株分泌TGF-β的活性,而此四個化合物對Jurkat細胞株的免疫調控機制仍須進一步探討。
The antler of Cervus elaphus L. (Lurong) was a famous Chinese traditional medicine. It was used for anti-inflammatory, anti-oxidation, anti-ageing, increasing the biosynthesis of proteins, anti-ulcerative, anti-tumor, and stimulation of sexual functions.
In the market, there had many different Lurong’s products, which were difficult to quantity the contents. In this program, crude extract of Lurong (CEEL-P) and three products from the market had been prepared and analyzed the protein contents by BCA assay and SDS-PAGE electrophoresis. The result indicated that the protein contents of three products (A、B、C) were 1 × 103 - 4.6 ×103 μg/mL, and two polypeptides were observed on SDS-PAGE electro- phoresis.
Iridoid was secondary metabolite that found in a wide variety of plants and in some animals. It exhibited many kind of bioactivities, including antibacterial, antioxidant, anti-inflammatory, analgesia, and hepatoprotective effects. In this research, we were reported the immunology effects of 12 iridoids: stilbericoside (1), 6-epi-stilbericoside (2), holmioside (3), thunaloside (4), 2-O-prime- verosylpaeonol (5), 6α-hydroxy- geniposide (6), gardenoside (7), ixoroside (8), geniposide (9), shanzhiside (10), genipin gentiobioside (11), and jasminoside B (12). In the cell viability assay, compound 1-12 showed no cytotoxicity on HT-29 cells at 1~10 µM. In IL-8 secretion assay, HT-29 cells were treated with compound 1-12 for 12, 24, and 48 hr at different concentration (1~10 µM), and IL-8 secretion was determined by ELISA IL-8 assay kit. The result indicated that compound 1-5 exhanced IL-8 secretion of HT-29 cells, and compound 6-12 inhibited IL-8 secretion of HT-29 cells. Compound 7, 10-12 showed the significant inhibition of IL-8 secretion of HT-29 cell lines. Therefore, HT-29 cells, treated by compound 7, 10-12, was detected the IL-10 secretion and TGF-β. The results showed that compound 7, 10-12 didn’t enhance the IL-10 secretion of HT-29 cells. However, Compound 7, 10-12 showed the signigicant enhance TGF-β secretion of HT-29 cells. The mechanism of immunomodulation of jurkat cells remain to be clarified.
中文摘要..............................................i
英文摘要..............................................ii
誌謝..................................................iv
目錄..................................................v
表目錄................................................viii
圖目錄................................................ix
第一篇 鹿茸胜肽成分研究...............................1
第一章 緒論...........................................1
第一節 鹿茸藥材介紹...................................1
第二節 研究目的.......................................4
第二章 文獻回顧.......................................5
第一節 成份回顧.......................................5
第二節 藥理與藥效回顧.................................8
第三節 專利檢索.......................................14
第三章 實驗材料與方法.................................16
第一節 實驗材料.......................................16
第二節 實驗方法.......................................19
第四章 實驗結果.......................................22
第五章 結論...........................................24
參考文獻..............................................25
第二篇 Iridoids免疫活性研究...........................28
第一章 緒論...........................................28
第一節 研究背景.......................................28
第二節 Iridoid簡介....................................30
第三節 研究目的.......................................31
第二章 文獻回顧.......................................32
第一節 Iridoids 相關藥理回顧..........................32
第二節 專利檢索.......................................36
第三章 實驗材料與方法.................................39
第一節 實驗材料.......................................39
第二節 實驗方法.......................................41
第四章 實驗結果.......................................44
第一節 細胞存活率測試結果.............................44
第二節 HT-29 細胞株IL-8分泌含量分析結果...............47
第三節 HT-29 細胞株IL-10分泌含量分析結果..............51
第四節 HT-29 細胞株TGF-β分泌含量分析結果..............53
第五章 結論...........................................55
參考文獻..............................................56
英文論文大綱..........................................59
個人簡歷..............................................64
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第二篇
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