鹿茸胜肽成分研究及iridoids免疫活性研究

Study on the protein of Cervus elaphus L. antler and the immunology assay of iridoids.

何欣欣(Xin-Xin He)

指導教授 ： 石麗仙

國立虎尾科技大學/電機資訊學院/光電與材料科技研究所/碩士(2009年)

摘要

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。

關鍵字

鹿茸 ； 環烯醚萜類 ； 介白素-8 ； 介白素-10 ； 轉化生長因子-β

並列摘要

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.

並列關鍵字

Cervus elaphus ； iridoid ； IL-8 ； IL-10 ； TGF-β

參考文獻

第一篇

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