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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 孫璐西 | |
dc.contributor.author | Yao-Yun Chang | en |
dc.contributor.author | 張耀云 | zh_TW |
dc.date.accessioned | 2021-06-16T23:34:09Z | - |
dc.date.available | 2012-08-01 | |
dc.date.copyright | 2012-08-01 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-07-27 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/65276 | - |
dc.description.abstract | 糖尿病是一種體內葡萄糖恆定失調的代謝性疾病,主要由於胰島素分泌或胰島素作用有缺陷,而影響血中葡萄糖進入肝臟、肌肉及脂肪等組織所引起的高血糖。脂肪與肌肉細胞皆為受胰島素調控來維持血糖恆定的主要組織。文獻指出,週邊組織攝入葡萄糖,對於血糖的恆定及清除餐後血糖是很重要的步驟。因此,評估細胞的葡萄糖攝取能力,在糖尿病相關研究中,扮演一個很基礎的角色。羅漢果的食用、藥用價值已有很悠長的歷史。除了甜味的特性之外,研究也發現羅漢果在體內與體外試驗中,均具有降血糖之活性。因此本次實驗目的為利用3T3-L1脂肪細胞之葡萄糖攝取試驗為篩選模式,找出羅漢果中可能具有降血糖活性之成分。結果顯示,羅漢果粗萃物能夠顯著提升脂肪細胞的葡萄糖攝取量,且具濃度效應。將粗萃物依極性區分後,發現以乙酸乙酯層的效果最佳。更進一步將乙酸乙酯層利用管柱層析分離後,在其次區分層中分離出S1、S2與S3;此三個樣品皆能顯著增加脂肪細胞的葡萄糖攝取能力。當單純以樣品處理時,S1具有最好的效果,然而與胰島素共同處理下,S1則降低了胰島素的活性。經由西方點墨法分析,發現S1本身可增加Akt磷酸化;S2的促進效果同時存在於單獨處理與有10 nM胰島素刺激的狀況。綜合以上結果,推測S1具有類似胰島素之作用,S2應有協同胰島素作用之活性。 | zh_TW |
dc.description.abstract | Diabetes mellitus is a metabolic disorder characterized by hyperglycemia resulting from defects in insulin secretion, insulin action, or both. Muscle and fat cells are both insulin-stimulated target tissues which regulate the blood glucose concentration. Glucose uptake in target tissues is a critical step in maintaining glucose homeostasis and clearing the postprandial glucose load. Therefore, evaluation of glucose uptake ability in cells plays a fundamental role in diabetes research. Siraitia grosvenori (SG), also called Luo-Han-Guo (LHG), is a fruit used in traditional Chinese medicine and also used as food additives. Besides its sweet taste, studies showed that LHG-extract exhibited an antidiabetic effect on both in vivo and in vitro trials. The aim of this study is to find the active compounds with hypoglycemic potential in LHG by glucose uptake assay in 3T3-L1 cells. LHG crude extract increased glucose uptake in a dose-dependent manner, and after solvent partition of crude extract, the ethyl acetate fraction showed the best glucose uptake activity. There were three components (S1, S2 and S3) separated from EA fraction by column chromatography, and all of them can significantly increase glucose uptake activity in 3T3-L1 adipocytes. Among them, S1 showed the best activity in basal glucose uptake. In combination with insulin, however, S1 slightly reduced insulin-stimulated glucose uptake. By western blot analysis, S1 enhanced the phosphorylation level of Akt. S2 was found to have a promotion effect on both basal and insulin-stimulated glucose uptake. In conclusion, S1 had an insulin-mimetic action and S2 showed synergistic effect with insulin in 3T3-L1 adipocytes. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T23:34:09Z (GMT). No. of bitstreams: 1 ntu-101-R99641034-1.pdf: 3354142 bytes, checksum: b161c2ccd0be0116023947321b63358d (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | 目錄
中文摘要………………………………………………………………………………I 英文摘要……………………………………………………………………………...II 縮寫表……………………………………………………………………………......III 目錄………………………………………………………………………………......IV 圖次……………………………………………………………………………........VII 表次……………………………………………………………………………..........IX 壹、前言……………………………………………………………………………......1 貳、文獻整理………………………………………………………………………......2 第一節、 材料介紹..………………………………………………………….....2 一、羅漢果簡介…………….…………………………………………….......2 二、羅漢果成分與生理活性……………………………….…………….......2 三、羅漢果之調節血糖功效…………………………….……………….......4 第二節、 糖尿病簡介………………………………….......................................7 一、葡萄糖代謝與血糖恆定…………………………………........................7 二、胰島素作用機制…………………………………....................................9 三、糖尿病定義及分類…………………………………..............................13 第三節、 3T3-L1脂肪細胞模式………………...………………………….....15 一、脂肪組織與葡萄糖恆定……………...……………………………......15 二、3T3-L1前脂肪細胞介紹……………..…………………………..........19 三、葡萄糖攝取試驗(Glucose uptake assay) ……………...………………20 四、3T3-L1脂肪細胞之葡萄糖攝取試驗…………………………………21 參、研究目的與實驗架構……………...…………………………………..…….......28 第一節、研究目的………………...…………………………………..…….......28 第二節、實驗架構……...……………………….……………………..……......29 肆、材料與方法……...…………………………………..……...................................31 第一節、實驗材料與儀器設備………..……......................................................31 一、 羅漢果………..……............................................................................31 二、 實驗細胞株………..……....................................................................31 三、 化學藥品與試劑…………………...……..…….................................31 四、 細胞實驗各培養溶液………..……....................................................34 五、 細胞實驗各種溶液配方……………………………………………..35 六、 層析材料………..……........................................................................38 七、 化學分析儀器設備………..……........................................................39 八、 細胞實驗儀器設備………..………………...….................................41 九、 實驗耗材………..……………………………....................................42 第二節、實驗方法………..……..........................................................................43 一、 羅漢果粗萃物之分離純化..................................................................43 (一) 羅漢果粗萃物之溶劑區分萃取.......................................................43 (二) 羅漢果粗萃物乙酸乙酯層(LHG-EA)之XAD-7 gel管柱層析…...43 1. 樣品前處理……………………………………………………….43 2. 管柱充填………………………………………………………….43 3. 樣品注入………………………………………………………….44 4. 沖提條件………………………………………………………….44 5. 薄層層析法(Thin layer chromatography, TLC) …………...…….44 (三) 羅漢果LHG-EA-F8之Sephadex LH-20 gel 管柱層析…………..45 1. 管柱充填………………………………………………………….45 2. 樣品注入………………………………………………………….46 3. 沖提條件………………………………………………………….46 4. 薄層層析法(Thin layer chromatography, TLC) …………...…….46 (四) 羅漢果LHG-EA-F8-7之製備型薄層層析(Preparative TLC) ……47 (五) 羅漢果LHG-EA-F8-7 spot 1 (S1)之silica gel管柱層析………….48 (六) S2之製備…………………………………………………………....49 二、 葡萄糖攝取試驗……………………………………………………..51 (一) 前脂肪細胞3T3-L1之培養………………………………………..51 (二) 細胞繼代及冷凍保存……………………………………………...51 (三) 前脂肪細胞3T3-L1之分化培養……………………………...…...52 (四) 葡萄糖攝取測定法………………………...……………………....52 三、 羅漢果調節葡萄糖攝取作用之探討……...………………………...53 四、活性成分分析...…………………………………….……………........56 (一) 高效能液相層析(HPLC) ………………………….…………........56 (二) 質譜之測定………………………………………………………...57 (三) 核磁共振光譜之測定……………………….…………..................57 第三節、統計分析...…………………………..…………….…………………..59 伍、結果與討論…………………………..…………….…………………………….60 一、羅漢果初萃物之分離純化………….…………………………….…..60 (一) 羅漢果粗萃物之溶劑區分萃取………….……………..…………60 (二) 羅漢果粗萃物乙酸乙酯層(LHG-EA)之XAD-7 gel管柱層析…...60 (三) 羅漢果LHG-EA-F8之Sephadex LH-20 gel 管柱層析…………..62 (四) 羅漢果LHG-EA-F8-7之製備型薄層層析(Preparative TLC)…….64 (五) 羅漢果LHG-EA-F8-7 spot 1(S1)之silica gel管柱層析…………..67 (六) S2之製備………………………………………………………….67 二、葡萄糖攝取試驗…….……………………………….………….…….69 (一) 脂肪細胞之分化培養………………………….……………..……69 (二) 羅漢果調節血糖活性之評估………….……………..……………72 三、羅漢果調節葡萄糖攝取作用之探討……………..……………….….89 四、活性成分分析………….……………..……………………………….91 (一) 高效能液相層析(HPLC) ………….……………..………………..91 (二) 質譜與核磁共振光譜之測定……………………………………...92 陸、結論………….…………..……………………………………………………...100 柒、參考文獻………….……………..……………………………………………...101 捌、附錄……………………………………………………………………………..108 口試Q&A……………………………………………………………………....114 | |
dc.language.iso | zh-TW | |
dc.title | 以脂肪細胞探討羅漢果中具調節血糖之活性成分 | zh_TW |
dc.title | Investigation on the Active Hypoglycemic Components of Siraitia Grosvenori by Glucose Uptake Assay in 3T3-L1 Adipocytes | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 黃青真,吳亮宜,魏國晉,何其儻 | |
dc.subject.keyword | 羅漢果,降血糖,葡萄糖攝取,3T3-L1脂肪細胞, | zh_TW |
dc.subject.keyword | Siraitia grosvenori,anti-diabetic,glucose uptake,3T3-L1 adipocytes, | en |
dc.relation.page | 118 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-07-27 | |
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 |
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