目錄…………………………………………………………………………I
圖目錄……………………………………………………………………IV
表目錄……………………………………………………………………VI
縮寫表……………………………………………………………………VII
謝誌………………………………………………………………………IX
中文摘要…………………………………………………………………XI
英文摘要…………………………………………………………………XIII
第壹章、前言………………………………………………………………1
第貳章、文獻探討…………………………………………………………3
第2-1節 紫外線…………………………………………………….…..4
2-1-1. 紫外線……………………………………………………….4
2-1-2. 紫外線的分類……………………………………………….5
2-1-3. 紫外線對人體健康的益處………………………………….6
2-1-4. 紫外線對人體的傷害……………………………………….7
第2-2節 自由基…………………………………………………………8
2-2-1. 自由基的介紹……………………………………………….8
2-2-2. 自由基的來源……………………………………………….8
2-2-3. 紫外線與自由基…………………………………………….9
2-2-4. 自由基種類………………………………………………….9
第2-3節 抗氧化酵素訊息的傳遞路徑………………………………..12
2-3-1. Nf2…………………………………………………………..12
2-3-2. Keap1………………………………………………………..12
2-3-3. ARE………………………………………………………….13
2-3-4. Nrf2的活化機制…………………………………………….14
第2-4節 細胞抗氧化的防禦系統……………………………………17
2-4-1. 酵素型抗氧化防禦系統…………………………………..18
2-4-2. 非酵素型抗氧化防禦系統………………………………...24
第參章、研究動機與實驗設計架構圖……………………………………...25
第3-1節 硏究動機……………………………………………………..26
第3-2節 實驗設計架構………………………………………………..27
第肆章、實驗材料與方法……………………… …………………………28
第4-1節 實驗材料……………………………………………………...29
第4-2節 實驗儀器……………………………………………………...32
第4-3節 實驗方法……………………………………………………...33
4-3-1. 細胞培養…………………………………………………...33
4-3-2. 存活率試驗………………………………………………...36
4-3-3. ROS產量測定………………………………………………38
4-3-4. 細胞毒性的測定…………………………………………...40
4-3-5. Comet assay彗星試驗………………………………………41
4-3-6. 細胞GSH含量測定………………………………………..44
4-3-7. 細胞凋亡試驗……………………………………………...46
4-3-8. 細胞總蛋白質萃取………………………………………...48
4-3-9. 細胞核與細胞質之蛋白質萃取…………………………...49
4-3-10. 蛋白質定量……………………………………………….51
4-3-11. 西方墨點分析…………………………………………….52
4-3-12. 免疫螢光染色…………………………………………….56
4-3-13. 轉染作用………………………………………………….58
4-3-14. 冷光酵素報導基因分析………………………………….60
4-3-15. RNA干擾…………………………………………………62
4-3-16. 統計分析………………………………………………….64
第伍章、實驗結果與圖表…………………………………………………65
第5-1節 ERG對HaCaT細胞的抗光氧化能力………………………66
5-1-1. ERG以及UVA對HaCaT細胞的細胞毒性影響…………66
5-1-2. ERG對UVA誘導活性氧物種含量之影響………………..66
5-1-3. ERG對UVA誘導的細胞膜損傷之影響…………………..67
5-1-4. ERG對UVA誘導的DNA損傷之影響……………………67
5-1-5. ERG對UVA所誘導之氧化傷害的影響…………………..68
5-1-6. ERG對UVA引發之細胞凋亡的影響……………………..68
第5-2節 ERG誘導之抗氧化系統的機制探討………………………..79
5-2-1. ERG對抗氧化酵素的影響…………………………………79
5-2-2. ERG活化Nrf2路徑探討……………………………………80
5-2-3. ERG影響Nrf2入核的表現………………………………..80
5-2-4. ERG對抗氧化酵素轉錄的影響……………………………81
5-2-5. 以siNrf2探討ERG的保護機制和Nrf2之間的關係……81
第陸章、討論………………………………………………………………95
第6-1節 ERG對於UVA刺激HaCaT細胞造成的細胞損傷之影響..97
第6-2節 ERG對抗氧化基因的表現之影響………………………….97
第6-3節 ERG與UVA影響Nrf2-Keap1 pathway……………………98
第6-4節 Nrf2對ERG所抑制UVA造成之光氧化傷害的影響……..98
第柒章、結論……………… …………………………………………..100
參考文獻 ………………….……………………………………………..102
| Identifer | oai:union.ndltd.org:TW/101CMCH5793001 |
| Date | January 1900 |
| Creators | 羅珩瑋, Heng-wei Luo |
| Contributors | 許游章, You-Cheng Hseu |
| Publisher | 中國醫藥大學, 藥用化妝品學系碩士班 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | 中文 |
| Detected Language | Unknown |
| Type | 碩士 |
| Format | 113 |
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