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| 研究生： |
方奕閔 Fang, Yi-Min |
|---|---|
| 論文名稱： |
茶籽萃取物之抗發炎活性評估 Anti-inflammatory Activity Evaluation of Tea Seed Extract |
| 指導教授： |
施玟玲
Shih, Wen-Ling |
| 學位類別： |
碩士 Master |
| 系所名稱： |
農學院 - 生物科技系 Department of Biological Science and Technology |
| 畢業學年度： | 109 |
| 語文別： | 中文 |
| 論文頁數： | 43 |
| 中文關鍵詞： | 茶籽萃取物 、脂多醣體 、抗發炎 、小鼠動物實驗 |
| 外文關鍵詞： | Tea Seed Extract, Lipopolysaccharides, Anti-inflammatory, Laboratory mouse |
| DOI URL： | http://doi.org/10.6346/NPUST202100081 |
| 相關次數： | 點閱：30 下載：0 |
| 分享至: |
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目錄
第一章 前言 1
1.1研究背景 1
1.2研究動機 1
1.3研究架構 2
1.4研究成果的重要性 2
第二章 文獻回顧 3
2.1茶籽萃取物的功能性及成分 3
2.2發炎(Inflammation) 3
2.3超氧化物歧化酶(Superoxide Dismutase；SOD) 3
2.4促炎細胞激素 4
2.5總抗氧化能力(Total Antioxidant Capacity；TAC) 4
2.6環氧化酶( cyclooxygenase；COX ) 5
2.7一氧化氮與一氧化氮合成酶(Nitric oxide synthase；iNOS) 5
2.8第一型血基質氧化酶( Heme oxygenase；HO-1) 5
2.9脂多醣體( Lipopolysaccharides；LPS ) 6
第三章 材料與方法 7
3.1實驗材料 7
3.1.1儀器設備 7
3.1.2藥品及緩衝溶液 8
3.1.2.1組織裂解液 8
3.1.2.2蛋白質電泳與西方墨點法緩衝溶液 8
3.1.2.3蛋白質電泳之膠體配置 9
3.1.2.4蛋白分子量標記 10
3.1.2.5抗體 10
3.1.2.6酵素聯結免疫分析 (Enzyme linked immunosorbent assay, ELISA) 10
3.1.2.7抗氧化活性分析試劑 11
3.1.2.8其他藥品 11
3.1.3細胞品系 12
3.2茶籽萃取粉末製作 12
3.2.1苦茶粕粉末萃取 12
3.2.2旋轉式減壓濃縮 12
3.2.3冷凍乾燥 12
3.3細胞培養 13
3.3.1 Dulbecco’s Modified Eagle’s medium製備 13
3.3.2細胞解凍 13
3.3.3冷凍細胞 13
3.3.4 細胞繼代 13
3.3.4.1 RAW264.7巨噬細胞繼代培養 13
3.3.4.2 NIH 3T3小鼠纖維母細胞繼代培養 14
3.3.5細胞計數 14
3.4實驗方法 14
3.4.1細胞毒性分析 14
3.4.1.1細胞毒性測試之MTT液體配置 15
3.4.2酵素連結免疫吸附分析法(Enzyme Linked Immunosorbent Assay；ELISA) 15
3.4.2.1酵素連結免疫吸附分析法之細胞前處理 15
3.4.3抗氧化活性分析 16
3.4.3.1 Total Antioxidant Capacity (TAC)分析 16
3.4.3.2 Superoxide Dismutase (SOD)分析 16
3.4.4西方墨點法分析(Western blot analysis) 16
3.4.4.1蛋白質定量 16
3.4.4.2組織裂解 16
3.4.4.3聚丙烯醯胺凝膠電泳(Sodium dodecyl sulfate polyacrylamide gel electrophoresis；SDS-PAGE) 17
3.4.4.4蛋白質轉漬及抗體分析 17
3.5動物實驗之抗發炎體內保護試驗 17
3.5.1分組 18
3.5.2茶籽萃取物與Lipopolysaccharides製備 18
3.5.3餵食 18
3.5.4血液採集 18
3.5.5統計分析 19
第四章 結果 20
4.1細胞毒性分析 20
4.2發炎相關因子IL-6、IL-1β、TNF-α分析 20
4.3總抗氧化能力分析Total Antioxidant Capacity (TAC) 21
4.4超氧化物歧化酶活性分析Superoxide Dismutase (SOD) 21
4.5西方墨點法(Western Blot) 21
4.6動物實驗肝臟組織解剖 22
第五章 討論與結論 23
第六章 附錄 26
第七章 參考文獻 33
圖目錄
圖 一、細胞存活率分析(MTT assay)。 26
圖 二、細胞發炎因子IL-6檢測。 27
圖 三、細胞因子TNF-α分析圖。 27
圖 四、細胞因子IL-6分析圖。 28
圖 五、細胞因子IL-1β分析圖。 28
圖 六、總抗氧化能力分析圖。 29
圖 七、超氧化物歧化酶活性分析圖。 29
圖 八、發炎因子COX-2蛋白表現量。 30
圖 九、發炎因子iNOS蛋白表現量。 30
圖 十、抗發炎因子HO-1蛋白表現量。 31
圖 十一、餵食茶籽萃取物肝臟解剖圖。 31
表目錄
表 一、BSA(Bovine Serum Albumin)標準曲線配置表(單位：µL) 32
表 二、茶籽萃取物樣本對NIH-3T3、RAW264.7細胞之IC50濃度。 32
表 三、動物實驗抗發炎之體內保護試驗流程圖。 32
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校外公開2026/02/03