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研究生: 林冠言
Lin, Kuan-Yen
論文名稱: 熱加工食品添加胺基酸緩解庫氏細胞發炎反應之研究
Amino acids addition in thermal processing of food alleviates Acrolein-induced inflammation of Kupffer cells
指導教授: 沈賜川
Shen, Szu-Chuan
丁俞文
Ting, Yu-Wen
吳瑞碧
Wu, Ruwi-Bi
口試委員: 丁俞文
Ting, Yu-Wen
吳瑞碧
Wu, Ruwi-Bi
吳忠信
WU, CHUNG-HSIN
黃文忠
HUANG, WEN-JUNG
沈賜川
Shen, Szu-Chuan
口試日期: 2022/08/30
學位類別: 碩士
Master
系所名稱: 營養科學碩士學位學程
Graduate Program of Nutrition Science
論文出版年: 2022
畢業學年度: 110
語文別: 中文
論文頁數: 72
中文關鍵詞: 丙烯醛胺基酸庫氏細胞NLRP3發炎小體
英文關鍵詞: Acrolein, Amino acids, Kupffer cell, NLRP3 inflammasome
DOI URL: http://doi.org/10.6345/NTNU202201857
論文種類: 學術論文
相關次數: 點閱:114下載:0
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  • 肝臟是人體最重要的器官之ㄧ,肝臟每天受到許多外源性及內源性的威脅,若長期處於發炎狀態,將會造成不可逆的功能受損,因此肝臟的健康與否對人體而言極為重要。丙烯醛是一種廣泛存在於人類生活中的有毒物質,空氣及某些食物中均含有丙烯醛,而在食品加工的過程中也會產生丙烯醛。丙烯醛一旦經人體攝入後,因為其親和能力強,容易與蛋白質、DNA、核酸等物質反應,使這些物質喪失其原先存在於人體中的能力;另一方面,丙烯醛對細胞的影響包括發炎、氧化壓力、內質網壓力…等。本研究探討梅納反應中間產物丙烯醛對肝臟庫氏細胞發炎之影響,以及以不同胺基酸與丙烯醛共同加熱,探討熱反應物是否可降低丙烯醛對於庫氏細胞的發炎反應。本研究首先以0、50、100、150、200μM濃度丙烯醛熱反應物對於庫氏細胞進行細胞毒性(MTT)測試。結果顯示50μM丙烯醛熱反應物對庫氏細胞大約有2成的致死率。其次,以甘胺酸、丙胺酸、擷胺酸、胱胺酸、麩胺醯胺、蘇胺酸以及麩胺酸與丙烯醛在高溫下迴流製備熱反應物,探討熱反應物對庫氏細胞發炎的效果。實驗結果發現,半胱胺酸與丙烯醛熱處理反應可以降低丙烯醛的殘留量並提高庫氏細胞的細胞存活率,且西方墨點法分析也發現,丙烯醛-半胱胺酸熱反應物可以降低丙烯醛熱反應物對庫氏細胞中發炎相關的IKKβ、Nf-κB、caspase-1及NLRP3等蛋白表現量。總結本研究證實半胱胺酸在迴流加熱下可以與丙烯醛產生反應,以降低丙烯醛對於庫氏細胞的發炎情形,此研究結果亦可以為後續研究丙烯醛於肝臟的發炎機制以及胺基酸於熱加工食品開發時的參考。

    第一章、前言 1 第二章、文獻回顧 2 第一節、肝臟(Liver) 2 一、肝臟的結構 2 二、肝臟的功能 3 三、肝臟與疾病 3 四、肝臟的免疫細胞 5 第二節、庫氏細胞 (Kupffer cell) 8 一、庫氏細胞的功能 8 二、庫氏細胞與發炎反應 8 三、庫氏細胞中的發炎小體NLRP3 9 四、NLRP3發炎體活化機轉 10 第三節、丙烯醛(Acrolein) 13 一、丙烯醛特性與來源 13 二、丙烯醛的代謝 14 三、丙烯醛對細胞的影響 15 四、丙烯醛與胺基酸 17 第四節、半胱胺酸 (Cysteine) 19 一、半胱胺酸的結構與特性 19 二、半胱胺酸與疾病 19 三、半胱胺酸與丙烯醛 20 四、食物中的半胱胺酸 20 第三章、研究動機與目的及實驗架構 22 第一節、研究動機與目的 22 第二節、實驗架構 23 第四章、實驗材料與方法 24 第一節、實驗藥品與儀器 24 一、實驗細胞 24 二、實驗藥品及試劑 24 三、儀器設備 26 第二節、實驗步驟與方法 28 一、實驗樣品配置 28 二、實驗藥品配置 28 三、實驗方法 29 四、統計分析 35 第五章、實驗結果 36 第一節、丙烯醛組及不同胺基酸組對於庫氏細胞的細胞毒性試驗 36 第二節、用高效液相層析儀分析樣品丙烯醛的殘留量 40 第三節、丙烯醛介入庫氏細胞培養液中細胞激素產量之影響 44 第四節、丙烯醛及半胱胺酸組別清除DPPH氧化自由基能力測定 46 第五節、丙烯醛介入庫氏細胞的發炎小體相關蛋白質表現量影響 48 第六章、討論 53 第一節、丙烯醛組及不同胺基酸組對於庫氏細胞的細胞毒性試驗 53 第二節、用高效液相層析儀分析樣品丙烯醛的殘留量 55 第三節、丙烯醛介入庫氏細胞培養液中細胞激素產量之影響 57 第四節、丙烯醛及半胱胺酸組別清除DPPH氧化自由基能力測定 58 第五節、丙烯醛介入庫氏細胞的發炎小體相關蛋白質表現量影響 59 第七章、結論 60 參考文獻 62

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