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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 謝淑貞(Shu-Chen Hsieh) | |
dc.contributor.author | Chung-Ching Lee | en |
dc.contributor.author | 李重慶 | zh_TW |
dc.date.accessioned | 2021-06-08T03:29:21Z | - |
dc.date.copyright | 2019-08-20 | |
dc.date.issued | 2019 | |
dc.date.submitted | 2019-08-15 | |
dc.identifier.citation | 衛生福利部。2018。106年國人死因統計結果。
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/21242 | - |
dc.description.abstract | 目前治療糖尿病的藥物中,有一類藥物為α-葡萄糖苷酶(α-glucosidase)抑制劑,可抑制α-葡萄糖苷酶將澱粉、寡糖或雙糖分解成單糖的活性,進而延緩糖類吸收,達到穩定餐後血糖的效果,而在天然物中也存在著各種不同形式的α-葡萄糖苷酶抑制劑,本實驗室先前已發現將中國橄欖(Canarium album (Lour.) Raeusch.)水萃物殘渣經甲醇萃取後之乙酸乙酯分層(COE)作為樣品,在小鼠巨噬細胞Raw 264.7具有抗發炎效果,在高飲食糖尿病大鼠可以降低血糖及改善脂肪代謝,所以本實驗繼續探討此萃取物的抗糖尿病機制。
本實驗以體外酵素抑制實驗作為篩選平台,發現中國橄欖萃取物具有抑制酵母α-葡萄糖苷酶的效果,於是以層析的方式純化萃取物中所含有的功效成分,並鑑定其結構。最後純化到2個可水解型單寧化合物(hydrolysable tannins) 1,2,3,6-tetra-O-galloyl-β-glucopyranose和chebulagic acid。在本實驗的條件下測得抑制酵母菌α-葡萄糖苷酶之IC50由低到高分別為1,2,3,6-tetra-O-galloyl-β-glucopyranose (0.44±0.07 μg/ml)、chebulagic acid (0.94±0.13 μg/ml)、COE (1.25±0.12 μg/ml)、acarbose (214.80±15.45 μg/ml)。再以Caco-2細胞酵素抑制實驗發現COE和所分離到的單寧化合物的確對人類α-葡萄糖苷酶有抑制效果,但在5 μg/ml濃度下只有COE效果跟acarbose效果相當,而2個單寧化合物則比acarbose效果差,代表COE中可能有其他活性成分。COE水解的產物主要為gallic acid、ellagic acid與其他兩個UV吸收最大值在262 nm和272 nm的未知化合物(可能為酚類化合物),表示COE含有大量的可水解型單寧化合物,而因為經強酸加熱水解後的COE不再具有抑制α-葡萄糖苷酶的活性,我們推測COE中其它具有活性的成分也是可水解型單寧。另一方面又發現添加牛血清蛋白後即減弱了COE和單寧化合物抑制α-葡萄糖苷酶之效果,顯示COE可能與其他蛋白有非專一性的結合。綜合以上實驗推測COE含有的可水解型單寧為抑制α-葡萄糖苷酶功效的主要來源。 | zh_TW |
dc.description.abstract | α-glucosidase inhibitor, one of the interference used for diabetes treatment, can inhibit the activity of α-glucosidase to decompose starch, oligosaccharide or disaccharide into monosaccharides, thereby delaying the absorption of sugar and stabilizing postprandial blood glucose. Various α-glucosidase inhibitors in nature exhibit the ability for modulating blood sugar. Our previous studies display that ethyl acetate fraction of methanol extract from water extract residues of Chinese olive (Canarium album (Lour.) Raeusch.) (COE) has anti-inflammatory effect on mouse macrophage Raw 264.7 cell, and can reduce blood sugar and attenuate metabolic dysfunction in diabetic rats under high fat diet challenge. We thus continue to explore the anti-diabetic mechanism of COE in this study.
Firstly, in vitro enzyme inhibition experiment was used as a screening platform to identify compounds in COE with the inhibitory effect on yeast α-glucosidase. Based on the results from platform screening, we separated compounds from COE by chromatography, and purified two hydrolysable tannins, 1,2,3,6-tetra-O-galloyl-β-glucopyranose and chebulagic acid. Our in vitro enzyme screening revealed that the IC50 to yeast α-glucosidase from low to high is 1,2,3,6-tetra-O-galloyl-β-glucopyranose (0.44±0.07 μg/ml), chebulagic acid (0.94±0.13 μg/ml), COE (1.25±0.12 μg/ml) and acarbose (214.80±15.45 μg/ml). We further used Caco-2 cell enzyme inhibition assay to investigate whether COE and the compounds we found have an inhibitory effect on human α-glucosidase, and result demonstrated that the inhibitory effect of COE was almost equivalent to acarbose at 5 μg/ml. However, the effect of two compounds were lower than the effect of acarbose, indicating there are other active compounds in COE. The hydrolyzed products are mainly gallic acid, ellagic acid and two other unknown compounds (UV absorption λmax are 262 nm and 272 nm, may be phenolic compounds), indicated that COE contains a large amount of hydrolysable tannins. Since strong acid with heating hydrolyzed COE no longer exhibited inhibitory activities, we propose that hydrolysable tannin is also the major contributor in other active fractions from COE. On the other hand, the addition of bovine serum albumin reduces the effect of COE and tannin compounds on inhibiting α-glucosidase, indicating that the active ingredients in COE may non-specifically bind other proteins. In conclusion, we declared that the hydrolysable tannins of COE may contribute to the inhibitory effect on α-glucosidase. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T03:29:21Z (GMT). No. of bitstreams: 1 ntu-108-R06641023-1.pdf: 5585358 bytes, checksum: 9fc60b2539971b3eeb92f5a543eab952 (MD5) Previous issue date: 2019 | en |
dc.description.tableofcontents | 謝誌 i
摘要 ii 目錄 vi 縮寫表 xi 第一章、文獻回顧 1 第一節、糖尿病 1 一、 診斷標準 1 二、 分類 2 三、 第二型糖尿病病因 3 四、 高血糖對人體的影響 3 五、 降血糖藥物 5 六、 α-葡萄糖苷酶(α-glucosidase) 7 七、 α-葡萄糖苷酶抑制劑 9 第二節、中國橄欖 12 一、功效 13 二、中國橄欖所含之二次代謝物 15 第二章、實驗架構 23 第一節、研究目的 23 第二節、實驗流程 24 第三節、純化流程 25 第三章、材料與方法 26 第一節、材料 26 一、中國橄欖水萃物殘渣甲醇萃取乙酸乙酯區分層 26 二、分離純化 26 三、酵素活性實驗 27 四、細胞培養 27 第二節、實驗方法 28 一、樣品分離純化 28 二、結構解析 29 三、酵母菌 α-葡萄糖苷酶活性實驗 29 四、酸水解實驗 30 五、蛋白質對抑制效果的影響實驗 30 六、Caco-2 α-葡萄糖苷酶活性實驗 31 七、統計分析 32 第四章、結果 33 第一節、COE對酵母菌α-葡萄糖苷酶的抑制效果 33 第二節、COE經Sephadex LH-20管住層析區分層的結果 33 第三節、COE-F41經Diaon HP-20分離結果 36 第四節、COE-F41-40%次分層 38 一、HPLC分離結果 38 二、成分探討 41 第六節、COE-F41-60%次分層 46 一、HPLC分離結果 46 二、成分探討 50 第七節、人類大腸癌細胞Caco-2酵素抑制效果 58 第八節、COE酸水解 59 第九節、蛋白質對COE抑制酵母菌α-葡萄糖苷酶的影響 64 第五章、討論 66 第一節、分析方法 66 第二節、有效成分 67 第三節、應用限制 69 第六章、結論 71 第七章、參考文獻 72 第八章、附錄 80 | |
dc.language.iso | zh-TW | |
dc.title | 探討中國橄欖水萃殘渣之甲醇萃取物乙酸乙酯區分層中的α-葡萄糖苷酶抑制成分 | zh_TW |
dc.title | Investigation of α-glucosidase inhibitory compounds in ethyl acetate fraction of methanol extract from water extract residues of Chinese olive (Canarium album L.) | en |
dc.type | Thesis | |
dc.date.schoolyear | 107-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 李宗徽(Tzong-Huei Lee) | |
dc.contributor.oralexamcommittee | 呂廷璋(Ting-jang Lu),郭靜娟(Ching-Chuan Kuo) | |
dc.subject.keyword | 糖尿病,α-葡萄糖??,中國橄欖,可水解型單寧, | zh_TW |
dc.subject.keyword | diabetes,α-glucosidase,Chinese olive (Canarium album (Lour.) Raeusch.),hydrolysable tannins, | en |
dc.relation.page | 92 | |
dc.identifier.doi | 10.6342/NTU201903489 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2019-08-16 | |
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 |
文件中的檔案:
檔案 | 大小 | 格式 | |
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ntu-108-1.pdf 目前未授權公開取用 | 5.45 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。