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研究生: 黃沛縈
Huang, Pei-Ying
論文名稱: 探討Lunasin在肥胖微環境下對RAW264.7細胞與EL-4 T細胞免疫反應之影響
The effects of Lunasin on immune responses of RAW264.7 and EL-4 T cells in the obese microenvironment
指導教授: 謝佳倩
Hsieh, Chia-Chien
口試委員: 林璧鳳 吳文勉 謝佳倩
口試日期: 2021/07/15
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: 2021
畢業學年度: 109
語文別: 中文
論文頁數: 113
中文關鍵詞: 肥胖Lunasin巨噬細胞T細胞細胞激素
英文關鍵詞: Obesity, Lunasin, Macrophages, T cells, Cytokines
DOI URL: http://doi.org/10.6345/NTNU202101233
論文種類: 學術論文
相關次數: 點閱:149下載:0
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  • 第一章 研究動機與假說 1 第二章 文獻探討 2 第一節 免疫系統 2 一、 先天性免疫反應 2 (一) 先天性免疫簡介 2 (二) 先天性免疫相關細胞 3 1. 單核球與巨噬細胞 3 2. 樹突細胞 4 二、 後天性免疫反應 4 (一) 後天性免疫簡介 4 (二) 後天性免疫相關細胞 5 1. T細胞 5 2. B細胞 6 三、 發炎反應 6 第二節 肥胖 8 一、流行病學調查 8 二、肥胖生理意義 8 三、肥胖與免疫 9 (一) 肥胖狀態下的發炎反應 9 (二) 肥胖與免疫細胞 10 1.巨噬細胞 10 2. T細胞 11 (三)肥胖與細胞激素 12 1. Leptin 12 2. Adiponectin 12 3. PAI-1(plasminogen activator inhibitor-1) 13 4. VEGF (vascular endothelial growth factor) 14 5. IL-6和 TNF-α 14 6. MCP-1 15 7. IL-1β 16 8. IL-2和IL-4 16 第三節 肥胖與代謝疾病 17 第四節 天然食物成分對免疫系統之調節 17 第五節 Lunasin 20 一、 Lunasin簡介 20 二、 Lunasin的消化吸收 20 三、 Lunasin的生理功能 20 (一) 抗氧化 20 (二) 抗發炎、免疫調節 21 (三) 抗癌 22 (四) 心血管保護作用 23 第三章 實驗設計 24 第一節 研究架構 25 第四章 實驗材料與方法 26 第一節 實驗材料 26 一、 實驗儀器設備 26 二、 拋棄式無菌耗材 26 第二節 實驗方法 27 一、 細胞培養 27 (一) RAW264.7巨噬細胞株培養 27 (二) RAW264.7巨噬細胞MTT細胞存活率測定 28 (三) 3T3-L1小鼠纖維母細胞培養 28 (四) 3T3-L1小鼠纖維母細胞分化為脂肪細胞 29 (五) EL-4 T細胞培養 30 二、 建立肥胖之體外發炎模式 31 (一) LPS誘發RAW264.7巨噬細胞之發炎模式 31 (二) Ad-CM誘發RAW264.7巨噬細胞之發炎模式 31 (三) TNF-α誘發3T3-L1脂肪細胞之發炎模式 31 三、 Lunasin處理對3T3-L1各分化階段上清液的發炎激素濃度之影響 31 (一) 細胞激素分析:酵素連結免疫吸附分析 (Enzyme-linked immunosorbent assay, ELISA) 31 四、Lunasin處理對肥胖微環境下巨噬細胞發炎反應、遷移及其表型極化之影響 34 (一) 細胞激素分析:ELISA 34 (二) 遷移試驗 (Migration assay) 35 (三) 基因表現量測定:逆轉錄聚合酶鏈式反應 (Reverse Transcriptase PCR, RT-PCR or qPCR) 37 五、Lunasin處理對肥胖微環境下巨噬細胞氧化壓力之影響 40 (一) 氧化壓力試驗 40 (二) 細胞活性測定 41 六、Lunasin處理對肥胖微環境下巨噬細胞吞噬功能之影響 41 (一) 吞噬活性試驗 (phagocytosis assay) 41 (二) 胞飲活性試驗 (pinocytosis assay,中性紅試驗) 42 七、Lunasin處理對EL-4 T細胞與3T3-L1脂肪細胞共培養細胞激素之影響 43 第五章 實驗結果 45 第一節 Lunasin處理對3T3-L1分化期間細胞激素分泌之影響 45 第二節 Lunasin處理對肥胖微環境下巨噬細胞發炎反應、遷移及其表型極化之影響 48 一、 Lunasin處理對RAW264.7細胞存活率的影響 48 二、 Lunasin處理對肥胖微環境下RAW264.7巨噬細胞其細胞激素分泌之影響 49 三、 Lunasin處理對肥胖微環境下RAW264.7巨噬細胞之遷移能力的影響 51 四、 Lunasin處理對肥胖模式下RAW264.7巨噬細胞其表型極化之影響 52 第三節 Lunasin處理對肥胖模式下RAW264.7巨噬細胞氧化壓力之影響 56 一、Lunasin處理對肥胖模式下RAW264.7巨噬細胞ROS (H2O2)生成之影響 56 二、 Lunasin處理對肥胖模式下RAW264.7巨噬細胞細胞活性之影響 57 第四節 Lunasin處理對肥胖模式下巨噬細胞吞噬功能之影響 62 一、胞吞試驗 62 二、胞飲試驗 64 第五節Lunasin處理對EL-4 T細胞與3T3-L1脂肪細胞共培養其細胞激素之影響 66 第六章 討論 72 第一節 Lunasin處理對3T3-L1分化期間細胞激素之影響 72 第二節 Lunasin處理對肥胖模式下RAW264.7巨噬細胞發炎反應、遷移能力及其表型極化之影響 72 第三節 Lunasin處理對肥胖模式下RAW264.7巨噬細胞氧化壓力之影響 74 第四節 Lunasin處理對肥胖模式下RAW264.7巨噬細胞吞噬功能之影響 74 第五節Lunasin處理對EL-4 T細胞與3T3-L1脂肪細胞共培養細胞激素之影響 75 第七章 結論 77 附錄 78 參考文獻 86

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