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研究生: 王致中
Wang, Zhi-Zhong
論文名稱: 太平洋榅桲果皮萃取物之抗氧化及抗發炎活性探討
Antioxidant and anti-inflammatory activity of Cassemango (Spondias dulcis.) peel extracts
指導教授: 廖遠東
Liaw, Eun-Tun
學位類別: 碩士
Master
系所名稱: 農學院 - 食品科學系所
Department of Food Science
論文出版年: 2022
畢業學年度: 110
語文別: 中文
論文頁數: 106
中文關鍵詞: 太平洋榅桲抗氧化抗發炎小鼠巨噬細胞脂多醣
外文關鍵詞: Spondias dulcis, antioxidant, anti-inflammatory, mouse macrophage, lipopolysaccharide
DOI URL: http://doi.org/10.6346/NPUST202200277
相關次數: 點閱:59下載:0
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  • 摘 要 II
    謝 誌 V
    目 錄 VI
    圖 目 錄 XII
    表 目 錄 XIV
    第一章 前言 1
    第二章 文獻回顧 3
    2.1. 太平洋榅桲 (SPONDIAS DULCIS) 3
    2.1.1. 簡介 3
    2.1.2. 生理活性 3
    2.1.3. 其他檳榔青屬相關植物之研究 5
    2.2. 氧化壓力之介紹 6
    2.2.1. 自由基生成途徑及來源 6
    2.2.1.1. 內源性自由基 6
    2.2.1.2. 外源性自由基 7
    2.2.2. 自由基種類 7
    2.2.2.1. 超氧化物 (Superoxide, O2•-) 7
    2.2.2.2. 一氧化氮 (Nitric oxide, NO) 7
    2.2.2.3. 羥基自由基 (Hydroxyl radical, OH•) 8
    2.2.2.4. 過氧自由基 (Peroxyl radical, ROO•) 8
    2.2.2.5. 過氧化氫 (Hydrogen peroxide, H2O2) 8
    2.2.2.6. 單線態氧 (Singlet oxygen, 1O2) 8
    2.2.3. 自由基與疾病 9
    2.3. 發炎反應 10
    2.3.1. 急性發炎與慢性發炎 10
    2.3.2. 巨噬細胞所引起之發炎反應 10
    2.3.3. 脂多醣誘導之發炎反應 11
    2.3.4. 發炎因子 11
    2.3.4.1. 白血球介素-6 (Interleukin 6, IL-6) 11
    2.3.4.2. 腫瘤壞死因子 (tumor necrosis factor-alpha, TNF-α) 12
    2.3.4.3. 誘導型一氧化氮合酶 (inducible nitric oxide synthase, iNOS) 12
    2.3.4.4. 環氧合酶-2與前列腺素E2 13
    2.4. 抗氧化劑 14
    2.4.1. 酵素性抗氧化劑 (Enzymatic antioxidant) 14
    2.4.1.1. 超氧歧化酶 (Superoxide dismutases, SOD) 14
    2.4.1.2. 過氧化氫酶 (Catalase, CAT) 15
    2.4.1.3. 榖胱甘肽過氧化酶 (Glutathione peroxidases, GPx) 15
    2.4.2. 非酵素性抗氧化劑(Non-enzymatic antioxidant) 15
    2.4.2.1. 榖胱甘肽 (Glutathione, GSH) 15
    2.4.2.2. 維生素C (vitamin C) 16
    2.4.2.3. 維生素E (Vitamin E) 16
    2.4.2.4. β-胡蘿蔔素 (β- carotene) 16
    2.4.3. 酚類化合物 (Phenolic compounds) 21
    2.4.3.1. 酚酸類 (Phenolic acids) 21
    2.4.3.2. 類黃酮 (Flavonoids) 22
    第三章 材料與方法 25
    3.1. 實驗設計與架構 25
    3.2. 實驗材料 28
    3.2.1實驗樣品 28
    3.2.2實驗藥品 28
    3.2.3. 儀器設備 29
    3.2.4. 小鼠巨噬細胞株 30
    3.3. 實驗方法 31
    3.3.1. 樣品製備 31
    3.3.2. 萃取物製備 31
    3.3.2.1. 熱水萃取 31
    3.3.2.2. 乙醇萃取 31
    3.3.3. 抗氧化成分含量分析 32
    3.3.3.1. 總酚含量測定 32
    3.3.3.2. 總類黃酮含量測定 32
    3.3.4. 抗氧化能力分析 33
    3.3.4.1. DPPH˙自由基清除能力試驗 33
    3.3.4.2. ABTS˙+自由基清除能力測定 33
    3.3.4.3. 還原能力測定 34
    3.3.5. 有效成分分析與鑑定-高效能液相層析儀 (HPLC) 35
    3.3.6. 細胞培養 38
    3.3.6.1. 培養基配製 38
    3.3.6.2. PBS (Phosphate buffered saline) 配製 38
    3.3.6.3. 細胞解凍及培養 38
    3.3.6.4. 細胞繼代 39
    3.3.6.5. 細胞冷凍 39
    3.3.6.6. 細胞計數 39
    3.3.6.7. 配製脂多醣 40
    3.3.7. 細胞存活率試驗 (MTT assay) 40
    3.3.8. 一氧化氮生成分析 41
    3.3.9. 細胞激素TNF-α、IL-6及PGE2之含量測定 41
    3.3.10. 西方墨點法 (Western blot) 43
    3.3.10.1. 細胞蛋白質萃取 43
    3.3.10.2. 蛋白質濃度之定量分析 43
    3.3.10.3. 聚丙烯醯胺膠體電泳 (SDS-PAGE) 43
    3.3.10.4. 轉漬 (Transfer) 44
    3.3.10.5. 封閉及一、二級抗體反應 44
    3.3.11. 統計分析 47
    第四章 結果與討論 48
    4.1. 萃取物製備 48
    4.2. 抗氧化成分含量 50
    4.2.1. 總酚含量 50
    4.2.2. 總類黃酮含量 51
    4.3. 抗氧化能力測定 53
    4.3.1. DPPH自由基清除能力測定 53
    4.3.2. ABTS自由基清除能力測定 54
    4.3.3. 還原能力 55
    4.4. 高效能液向層析儀 (HPLC)酚類化合物之分析結果 60
    4.5. 相關性分析 69
    4.5. 細胞試驗 71
    4.5.1. 細胞存活率試驗 71
    4.5.2. 太平洋榅桲果皮萃取物對RAW 264.7小鼠巨噬細胞產生一氧化氮之影響 74
    4.5.3. 太平洋榅桲果皮萃取物對RAW 264.7小鼠巨噬細胞促發炎細胞因子之影響 77
    4.5.3.1. 太平洋榅桲果皮50%乙醇萃取物對IL-6生成之影響 77
    4.5.3.2. 太平洋榅桲果皮50%乙醇萃取物對TNF-α生成之影響 78
    4.5.3.3. 太平洋榅桲果皮50%乙醇萃取物對PGE2生成之影響 79
    4.5.4. 太平洋榅桲果皮萃取物對RAW 264.7 小鼠巨噬細胞發炎因子蛋白表現量之影響 83
    4.5.4.1. 太平洋榅桲果皮50%乙醇萃取物對iNOS蛋白表現之影響 83
    4.5.4.2. 太平洋榅桲果皮50%乙醇萃取物對COX-2蛋白表現之影響 84
    第五章 結論 87
    第六章 參考文獻 88

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