臺灣博碩士論文加值系統

燕窩（EBN）為16世紀時，做為御膳供奉给皇上，其特色在於很少有食用上的禁忌，可以長期服用做滋補。燕窩醫療功效在中國古代醫學著作中一直有被詳細記載，在傳統醫學中，燕窩被記載有養陰潤燥、益氣補中、治虛損、咳血、久痢、止咳化痰的效果。根據前人研究顯示從燕窩中提取的成分能刺激表皮生長因子（EGF），使得細胞增殖以及提升免疫力。近期更有研究表示，如果將燕窩以沸水處理後，能夠促進角膜細胞的增殖以及傷口癒合。至目前為止，所有的研究證明，將燕窩樣品溶於水後，萃取出其成分有利於免疫力的提升。
本研究中以SDS-PAGE證實所使用的燕窩為純正的燕窩，經SDS-PAGE分析，顯示燕窩樣品在50 kDa的100 kDa和150 kDa的位置具有蛋白質條帶；由Warren assay測試後，結果顯示本研究所使用的燕窩樣品約含有10％ 唾液酸；以Kjeldahl method測定其粗蛋白含量，顯示出其蛋白質含量約50％。參照前人研究的結果可判定本研究所使用的燕窩樣品為真正的純燕窩。將燕窩樣品以酵素進行水解，接著以P-2 Gel進行管柱層析以去除小分子溶質後，以周邊血液單核球細胞做免疫測試，發現燕窩水解物的樣品在提升細胞媒介的免疫能力上，效果不大；相對的，其對於在提升促發炎反應的細胞激素上則效果相當強且顯著，而其中的機制值得進一步探討。本研究已建立燕窩樣品的最佳水解條件，並以周邊血液單核球細胞試驗對水解燕窩進行初步的免疫調節能力的測試。

Edible Bird’s Nest (EBN) was a significant item in the cuisine and pharmacy of the Emperors of China during the 16th century. EBN has been known for its beneficial effects in promoting health in the Chinese for hundreds of years. In traditional Chinese medicine (TCM), EBN is believed to promote the wellbeing of multiple organs and body systems. Previous study showed that extracts from EBN stimulated epidermal growth factor (EGF) and improved the immune function and cell proliferation. Recently, hot-water extraction of EBN showed the ability to promote cornea cells proliferation and cornea wound healing. To date, all the previous studies demonstrated beneficial effects of the EBN extracts prepared by water extraction.
Bird's nest used in this study had been proved to be a pure bird’s nest by SDS-PAGE, sialic acid content and protein content. Qualification analyses were conducted by SDS-PAGE, Warren assay and Kjeldahl method to authenticate the EBN sample. It was found to possess three bands at about 50 kDa, 100 kDa and 150 kDa, possess a sialic acid content about 10% of dry weight, and possess a protein content about 50% of dry weight. According to the criteria on previous studies, the EBN sample was considered to be a real and pure bird’s nest. Subsequently, the EBN sample was applied to enzyme digestion and then separated by Bio-gel P2 for size exclusion chromatography. In PBMC immunomodulation test, it was found the hydrolysate of bird's nest had effects on enhancing T-cell mediated immunity, but is not strong enough; while whose inflammatory effects were very strong and significant. The present study aimed to find the optimal hydrolysis condition for EBN by plant enzymes and to know whether the basic components of EBN hydrolysate could induce immune response or not.

目錄
第一章　背景介紹 1
1.1 燕窩 1
1.2 多醣體 5
1.3 多醣體對免疫調節的研究 7
1.4 免疫反應 8
1.5 發炎反應 8
1.6 周邊血液單核球細胞 9
1.7 細胞存活率分析 10
1.8 脂多醣體 10
1.9 植物血凝素 11
1.10 細胞激素 11
1.11 實驗目的 18
1.12 實驗架構 19
第二章　材料與方法 20
2.1 蛋白質分解酵素之活性分析 20
2.2 SDS-PAGE樣品製備 21
2.3 SDS-PAGE分析 21
2.4 樣品水解程度分析 22
2.5 水解條件併入後製程之整體製程之大量試驗 22
2.6 蛋白質含量分析 23
2.7 總糖含量分析 24
2.8 唾液酸含量分析 24
2.9 膠體過濾層析法 25
2.10高效能液相層析分析 26
2.11周邊血液單核球細胞分析 26
第三章　結果與討論 27
3.1 酵素活性測試 27
3.2 燕窩水解程度分析 27
3.3 燕窩粗蛋白含量分析 28
3.4 燕窩總糖含量分析 28
3.5 燕窩唾液酸含量分析 28
3.6 膠體過濾層析法 29
3.7 高效能液相層析分析 29
3.8周邊血液單核球細胞分析 30
參考文獻 33
附錄 59

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