記錄 編號 12713 狀態 G0499396010 助教 查核 建檔完成 索書 號 查核完成 學校 名稱 輔仁大學 系所 名稱 食品科學系 舊系 所名 稱 學號 499396010 研究 劉乙燕 生(中) 研究 Yi-Yen Liu 生(英) 論文 名稱 (中) 論文 名稱 (英) 自植物乳桿菌 TWK10 發酵豆奶中分離純化調節血壓之保健成分 Isolation and Purification of Functional Ingredients from Soy Milk Fermented with Lactobacillus plantarum TWK10 to Improve hypertension 其他 題名 指導 教授 (中) 蔡宗佑 潘子明 指導 教授 (英) 校內 全文 開放 日期 2017.8.14 校外 全文 2017.8.14 開放 日期 全文 不開 放理 由 電子 全文 同意 送交 國圖. 國圖 全文 2017.8.14 開放 日期. 檔案 說明 書名 摘要 謝誌 目錄 內文 參考文獻 電子 全文 01 02 03 04 05 06 學位 類別 碩士 畢業 學年 度 100 出版 年 101 語文 別 中文 關鍵 豆奶 高血壓 乳酸菌 血管收縮素轉換酶 人類臍靜脈內皮細胞 字(中) 關鍵 字(英) 摘要 (中) 豆奶在亞洲國家中是一個有益健康且可廣泛取得蛋白質飲食來源。文獻 指出,攝食黃豆製品有助於預防心血管疾病與癌症之風險。本研究之目 的旨在分離純化 Lactobacillus plantarum TWK10 (TWK10) 發酵豆奶之生理 活性成份,並探討其對血管收縮素轉換酶 I (angiotensin-I converting enzyme, ACE) 活性與一氧化氮 (nitric oxide, NO) 調節血管舒張因子之影 響。首先以管柱層析方式純化 TWK10 發酵豆奶水萃物之活性成份,並 藉由核磁共振光譜法 (nuclear magnetic resonance, NMR) 進行純化物鑑定並 測定其生理活性。結果顯示,TWK10 發酵豆奶水萃分離物 2-2 與 2-3 具 有抑制 ACE 之活性,最大半抑制濃度分別為 7.26 ug/mL 與 9.19 ug/mL。 此外,以濃度為 100 ug/mL 之分離物 2-2 與 2-3 處理人類臍靜脈內皮細胞 (human umbilical vein endothelial cell, HUVEC),可分別增加 1.36 與 1.35 倍 之 NO 產量。分離物 2-2 與 2-3 可再純化出 2-2A、2-2B、2-3A 與 2-3B 等 四個純化物,其中以 2-2A 與 2-3B 具有較高之抑制 ACE 活性,半抑制濃 度為 1.35 ug/mL 與 2.37 ug/mL;四個純化物無論處理 HUVEC 12 或 24 小 時皆可促進 NO 生成與清除超氧陰離子之活性。以 2-2A 處理 HUVEC 細 胞可促進前列腺素 E2 (prostaglandin E2, PGE2) 增加。以 2-2A 處理 HUVEC 12 小時,其內皮型一氧化氮合成酶 (endothelial nitric oxide synthase, eNOS) 表現與控制組之表現量無差異;以 2-3B 處理 12 小時後 eNOS 與環氧合酶-2 (cyclooxygenase-2, COX-2) 之表現量則高於控制組, 但高濃度之 2-2A 則會使 COX-2 表現量降低。藉由 NMR 與 LC-MS/MS 等 數據結果鑑定確認純化物 2-2A 之 IUPAC 命名為 (5-(2,4-dioxo-3-(1,2,3trihydroxypropyl)-3,4-dihydropyrimidin-1(2H)-yl)-3,4- dihydroxytetrahydrofuram2yl)methyl trihydrogen diphosphate。綜合上述,TWK10 乳酸菌發酵豆奶具 有抑制 ACE 活性與促進血管舒張之多功能保健成分,可開發為改善高血 壓及其衍生性疾病之保健食品。 摘要 (英) 論文 目次 參考 第一章 緒言 1 第二章 文獻回顧 2 2.1 益生菌與乳酸菌 2 2.1.1 益生菌 2 2.1.2 乳酸菌 2 2.1.3 乳酸菌之胞外多醣 3 2.1.4 乳酸菌對基質中蛋白質變化 之影響 3 2.1.5 已發表之乳酸菌具有之生理功效 3 2.2 發酵豆製品 5 2.2.1 黃豆 5 2.2.2 促進消化吸收 5 2.2.3 改善風味 6 2.3 高血壓 8 2.3.1 高血壓發 生 10 2.3.1.1 原發性高血壓 10 2.3.1.2 續發性高血壓 10 2.3.2 高血壓之預防 及治療 12 2.4 調節血壓機制之回顧 15 2.4.1 血管收縮素轉化酶抑制劑 15 2.4.1.1 特性及作用功能 15 2.4.1.2 抑制作用機轉 15 2.4.2 血管收縮素轉換 酶抑制物來源 17 2.4.3 血管收縮素受體阻斷劑 23 2.4.3.1 特性與作用機轉 23 2.4.3.2 ARB 之作用 24 2.5 血管收縮及舒張因子 32 2.5.1 血管構造 32 2.5.1.1 動脈 32 2.5.1.2 靜脈 32 2.5.1.3 微血管 33 2.5.2 血管收縮因子 33 2.5.3 內皮衍生舒張因子 34 第三章 研究目的與試驗架構 39 第四章 試驗材料 41 4.1 試驗菌株 41 4.2 試驗細胞 41 4.3 化學藥品 42 4.4 儀器設備與耗材 44 4.4.1 儀器設 44 4.4.2 耗材 45 4.5 試驗方法 46 4.5.1 乳酸菌發酵豆奶製備 46 4.5.2 發酵豆奶水萃取之製備 46 4.5.3 萃取物之純化分離 46 4.5.4 ACE 活性抑制分析 47 4.5.5 HUVEC 活化 49 4.5.6 HUVEC 培養與繼代 49 4.5.7 一氧化氮生成量測定 49 4.5.8 超氧陰離子試驗 50 4.5.9 前列腺素 E2 試驗 51 4.5.10 eNOS 與 COX-2 表現之測定 52 4.6 統計分析 55 第五章 結果與討 論 56 5.1 分離純化發酵豆奶 56 5.2 探討發酵前後與不同批次發酵豆奶之 有效成份分佈 56 5.3 抑制 ACE 活性 62 5.3.1 發酵豆奶水萃分離物抑制 ACE 活性 62 5.3.2 發酵豆奶水萃分離純化物抑制 ACE 活性 63 5.4 促進 HUVEC 之一氧化氮生成量之探討 70 5.5 抑制 HUVEC 中超氧陰離子生成 量之探討 76 5.6 促進 HUVEC 之前列腺素 E2 生成量之探討 82 5.7 HUVEC 之 eNOS 與 COX-2 表現量之探討 86 5.8 L. plantarum TWK10 發酵豆奶水 萃純化物之鑑定 90 第六章 結論與未來展望 101 第七章 參考文獻 102 丁予安 (1999) 臨床高血壓學第三版。藝軒圖書出版社,台北,台灣。 李 文獻 福臨 (2000) 乳酸菌分類之研究近況。食品工業 32:36-42。 林棟雍,張簡 玫芳,楊政儒 (2005) 培養溫度對於乳酸菌胞外多醣產量、單醣成分與分 子量之影響。華岡農科學報 15:21-30。 廖啟成 (1998) 乳酸菌之分類及應 用。食品工業 30:1-10。 潘子明 (2008) 乳酸菌的保健功效。健康世界 266:41-66。 Abraham, D, Handler, C, Dashwood, M, Coghlan, G (2008) Vascular complications in human disease : mechanisms and consequences. 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Am J Hypertens 21:72-77. 論文 頁數 附註 全文 點閱 次數 資料 建置 時間 2012/8/14 轉檔 日期 2012/08/15 全文 檔存 取記 錄 499396010 2012.8.14 17:58 140.136.182.98 new 01 499396010 2012.8.14 17:58 140.136.182.98 new 02 499396010 2012.8.14 17:59 140.136.182.98 new 03 499396010 2012.8.14 18:00 140.136.182.98 new 04 499396010 2012.8.14 18:00 140.136.182.98 new 05 499396010 2012.8.14 18:00 140.136.182.98 new 06 異動 記錄 C 499396010 Y2012.M8.D14 17:53 140.136.182.98 M 499396010 Y2012.M8.D14 17:55 140.136.182.98 M 499396010 Y2012.M8.D14 17:55 140.136.182.98 M 499396010 Y2012.M8.D14 17:56 140.136.182.98 M fosc2513 Y2012.M8.D14 18:19 140.136.182.98 M 499396010 Y2012.M8.D15 11:06 140.136.182.97 M FOSC2513 Y2012.M8.D15 11:49 140.136.174.68 M FOSC2513 Y2012.M8.D15 11:49 140.136.174.68 M FOSC2513 Y2012.M8.D15 11:49 140.136.174.68 M FOSC2513 Y2012.M8.D15 11:49 140.136.174.68 M FOSC2513 Y2012.M8.D15 11:49 140.136.174.68 M FOSC2513 Y2012.M8.D15 11:49 140.136.174.68 M FOSC2513 Y2012.M8.D15 11:49 140.136.174.68 M FOSC2513 Y2012.M8.D15 11:49 140.136.174.68 M 030540 Y2012.M8.D15 12:38 140.136.208.63 M 030540 Y2012.M8.D15 13:11 140.136.208.244 I 030540 Y2012.M8.D15 13:13 140.136.208.244