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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 黃青真(Ching-Jang Huang) | |
dc.contributor.author | Wei-Ti Yang | en |
dc.contributor.author | 楊惟蒂 | zh_TW |
dc.date.accessioned | 2021-05-20T21:54:24Z | - |
dc.date.available | 2012-07-30 | |
dc.date.available | 2021-05-20T21:54:24Z | - |
dc.date.copyright | 2010-07-30 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-07-26 | |
dc.identifier.citation | 吳美嬌(2000)。以巨噬細胞株為模式探討促進或抑制前列腺素E2生成的食物或食物成分。國立台灣大學農業化學研究所碩士論文。
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/10737 | - |
dc.description.abstract | Metabolic syndromes predict a high risk of cardiovascular diseases and type 2 diabetes. Insulin resistance and abnormal regulation of glucose homeostasis are major part of metabolic syndromes. Tissue-specific knockouts of the insulin receptor in liver and islet beta cells in mice resulted in impaired plasma glucose control, indicating that both liver and islet beta cells are indispensible in the regulation of body glucose homeostasis and metabolism. Bitter gourd (BG, Momordica charantia) is known for its hypoglycemic activity, but the active compounds and molecular target have not been clearly understood. This study aimed at examining effects of wild BG (Hualien No.4) water extract and fraction on the glucose uptake of hepatocytes (FL83B hepatic cell line) and on the insulin secretion of islet beta cells (RIN-m5F and HIT-T15 cell line).
Glucose uptake by FL83B cells was measured by the reduction of glucose concentration in the culture media after cells were treated with insulin (positive control) or MC extract/fractions for 16 or 20 hrs. Insulin secretion was measured after RIN-m5F and HIT-T15 cells were treated with 10mM glucose (positive control) or MC extract/fractions for 1 hour. Insulin secretion was also measured in HIT-T15 cells pretreated with 100uM palmitic acid for 48 hours to blunt the response of insulin secretion to glucose. The results shows that the P-fraction (containing insulin-like peptide), the WE (water extract) and its low MW (<3kD) fractions (WES) of MC significantly enhanced the glucose uptake of FL83B and the insulin secretion in both of the two beta cell lines (p<0.05). WE and WES were further hydrolyzed by incubation with β-glucosidase. The hydrolyzed products were sequentially extracted by ethyl acetate (EA) and butanol(B). The two extracts of WE also significantly enhanced glucose uptake of FL83B and insulin secretion of HIT-T15 (P<0.05). In addition, the EA extract of theβ-glucosidase hydrolyzed WES also increased the glucose uptake of FL83B and the insulin secretion in both of the beta cells (P <0.05). It was found that one fraction obtained from the prepared HPLC separation of the EA extract of the β-glucosidase hydrolyzed WES significantly improved the impaired insulin secretion of HIT-T15 pretreated with palmitic acid. In conclusion, these results suggest that the hypoglycemic effect of Hualien No. 4 wild BG is attributed, at least in part, to its active component(s) that enhanced the glucose uptake of liver and the insulin secretion of islet beta cells. | en |
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dc.description.tableofcontents | 中文摘要 i
英文摘要 ii 縮寫對照表iv 目錄 I 表目錄 VIII 圖目錄 IX 第一章、序論 1 第一節 研究動機與目的 1 第二節 文獻回顧 1 一、 代謝症候群 1 (一) 代謝症候群發展與危險因子指標 1 (二) 我國代謝症候群現況 2 二、 胰島beta細胞與胰島素 3 (一) 胰臟與胰島beta細胞 3 (二) 胰島素生成過程 3 (三) 葡萄糖刺激胰島素分泌(GSIS) 5 (四) 胰島素分泌障礙 7 三、 第二型糖尿病 9 (一) 胰島素阻抗 9 (二) 第二型糖尿病 10 (三) 糖尿病與肝臟 11 四、 食材樣品-山苦瓜 14 第三節 實驗假說與實驗設計 17 一、 實驗假說 17 二、 實驗設計 17 (一) FL83B小鼠肝臟細胞 17 1. 葡萄糖汲取模式建立 17 2. 樣品效用評估 17 (二) HIT-T15倉鼠胰島細胞株/RIN-m5F大鼠胰島細胞瘤之細胞株 17 1. 正常beta細胞 17 2. 以棕櫚酸誘發胰島素分泌障礙之胰島beta細胞 17 第二章、小鼠肝細胞株 FL83B葡萄糖汲取試驗 18 第一節 前言 18 第二節 細胞培養系統 20 第三節 山苦瓜萃取物/區分物實驗材料製備與分析 22 一、 實驗材料製備 22 (一) 山苦瓜水萃物(WE)製備 22 (二) 山苦瓜水萃物小分子區分物(WES)製備 23 (三) 山苦瓜水萃物小分子區分物經 b-glucosidase酵素水解之區分物製備(We-E, Se-E, We-B, Se-B, Wn-E, Sn-E) 23 (四) P-fraction(Pf)製備 24 二、 製備級HPLC分析 24 (一) 樣品前處理 24 (二) 製備式HPLC儀器設備 24 (三) 製備式HPLC分析流程與儀器設定 24 三、 實驗方法 25 (一) FL83B小鼠肝臟細胞 25 (二) 葡萄糖濃度測定 25 1. 原理 25 2. 實驗步驟 25 (三) 蛋白質測定-Bicinchoninic acid procedure (BCA) 26 1. 原理 26 2. 試劑與器材 26 3. 實驗步驟 26 (四) FL83B最佳胰島素作用時間與作用濃度測試 26 1. 實驗流程 26 2. 實驗步驟 27 (五) 不同山苦瓜區分物對FL83B細胞葡萄糖汲取影響 28 1. 實驗流程 28 2. 實驗步驟 28 四、 統計分析 28 第四節 實驗結果 29 一、山苦瓜水萃物與小分子區分物經酵素水解後EA萃物結果 29 二、山苦瓜區分物葡萄糖含量測定 29 三、FL83B肝細胞株葡萄糖汲取模式建立 30 (一) 最佳胰島素作用時間 30 (二) 最佳胰島素作用濃度 30 四、山苦瓜水萃物及其區分物處理FL83B之培養液葡萄糖減少量試驗 30 (一) 山苦瓜水萃物(WE)與其有或無酵素水解區分物(We-E, We-B, Wn-E) 30 (二) 山苦瓜水萃物之小分子區分物(WES)與其有或無酵素水解區分物(Se-E, Se-B, Sn-E) 31 (三) 山苦瓜水萃物含植物胰島素之多肽P-fraction 31 (四) 以製備級HPLC分離水萃物酵素水解乙酸乙酯區分物(WRP1-4) 32 (五) 以製備級HPLC區分之小分子區分經酵素水解乙酸乙酯區分物(SRP1-4) 32 第五節 討論 41 一、模式建立 41 (一) 培養液 41 (二) 取樣時間與胰島素濃度 41 二、山苦瓜水萃物與其區分物對FL83B細胞汲取葡萄糖之影響 42 (一) 山苦瓜水萃物與其區分物所含葡萄糖量對FL83B細胞汲取葡萄糖的影響 42 (二) 酵素水解對山苦瓜水萃物與小分子區分物於FL83B細胞汲取葡萄糖的影響 42 (二) We-E、WRP2與SRP4於高濃度減緩FL83B細胞汲取葡萄糖 43 三、山苦瓜水萃物與其區分物降低FL83B肝細胞株培養液葡萄糖含量可能原因探討 43 (一) 促進肝細胞汲取葡萄糖 43 1. 活化肝臟中PPARg以促進glucokinase(GK)基因表現 43 2. 類胰島素成分活化胰島素訊息傳遞路徑 44 (二) 抑制糖質新生作用 45 1. 直接抑制糖質新生相關基因與蛋白質表現 45 2. 活化AMPK(AMP-activated protein kinase)路徑以抑制糖質新生相關基因表現 45 第六節 結論 47 第三章、胰島beta細胞株胰島素分泌試 48 第一節 前言 48 第二節 細胞培養系統 48 一、細胞株 48 (一) RIN-m5F細胞株 48 (二) HIT-T15細胞株 49 二、試劑與器材 49 三、實驗方法 52 (一) 不同濃度葡萄糖下,RIN-m5F細胞與HIT-T15細胞之胰島素分泌試驗 52 1. 實驗流程 52 2. 實驗步驟 52 (二) 樣品處理 RIN-m5F與HIT-T15 胰島素分泌改變量分析 52 (三) 樣品處理對經棕櫚酸(PA)處理之 RIN-m5F與HIT-T15細胞胰島素分泌影響 53 1. 實驗流程 53 2. 實驗步驟 53 (四) 細胞存活率測定-MTT染色法 54 1. 原理 54 2. 實驗步驟 54 (五) 細胞培養液胰島素分析套組 54 四、統計分析 54 第三節 實驗結果 57 一、 Glucose stimulated insulin secretion 57 二、山苦瓜水萃物及其區分物處理正常狀態beta細胞之胰島素反應試驗 57 (一) 山苦瓜水萃物(WE)其有或無酵素水解區分物(We-E,We-B, Wn-E) 57 (二) 山苦瓜水萃物之小分子區分物(WES)與其有或無酵素水解區分物(Se-E, Se-B, Sn-E) 58 (三) 山苦瓜水萃物含植物胰島素之多肽P-fraction(Pf) 58 (四) 以製備級HPLC分離水萃物酵素水解乙酸乙酯萃物(WRP1-4) 59 (五) 以製備級HPLC區分之小分子區分經酵素水解乙酸乙酯萃物(SRP1-4) 59 三、棕櫚酸誘發beta細胞GSIS不良 59 四、山苦瓜水萃物及其區分物處理GSIS不良beta細胞之胰島素反應試驗 60 (一) 山苦瓜水萃物(WE)其有或無酵素水解區分物(We-E,We-B, Wn-E)60 (二) 山苦瓜水萃物之小分子區分物(WES)與其有或無酵素水解區分物(Se-E, Se-B, Sn-E) 60 (三) 山苦瓜水萃物含植物胰島素之多肽P-fraction(Pf)60 (四) 以製備級HPLC區分之小分子區分經酵素水解乙酸乙酯萃物(SRP1-4) 60 第四節 討論 73 一、 HIT-T15與RIN-m5F細胞株GSIS比較 73 二、 山苦瓜水萃物與其區分物對正常與胰島素分泌不良beta細胞分泌胰島素的影響 74 (一) 樣品所含葡萄糖促進胰島素分泌 74 (二) 胰島beta細胞膜上鉀離子通道或鈣離子通道的影響 74 三、棕櫚酸對胰島beta細胞分泌胰島素之影響 75 第五節 結論 77 第四章、綜合討論與總結論 78 第一節 綜合討論 78 第二節 總結論 81 第五章、參考文獻 82 | |
dc.language.iso | zh-TW | |
dc.title | 山苦瓜水萃物暨其區分物對肝細胞汲取葡萄糖及胰島beta細胞分泌胰島素之影響 | zh_TW |
dc.title | The Effects of Momordica charantia Linn. Water Extracts and
Fractions on Glucose Uptake of Hepatic Cell Line and Insulin Secretion of Islet Beta Cell Line | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 林璧鳳(Bi-Fong Lin),蔡帛蓉(Po-Jung Tsai),趙蓓敏(Pei-Min Chao),呂紹俊(Shao-Chun Lu) | |
dc.subject.keyword | 苦瓜水萃物,糖尿病,肝細胞株,胰島β細胞株, | zh_TW |
dc.subject.keyword | water extract of Momordica charantia,type 2 diabetes,hepatic cell line,islet beta cell line, | en |
dc.relation.page | 92 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2010-07-27 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
顯示於系所單位: | 微生物學科所 |
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