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研究生: 林冠宇
Lin, Kuan-Yu
論文名稱: 杜鵑花酸提升間葉幹細胞旁分泌效應調節抗老化因子miRNA-7850-5p 延緩皮膚衰老
Azelaic acid increases paracrine effect on mesenchymal stem cells to regulate anti-senescent marker miRNA-7850-5p in aging skin
指導教授: 陳冬生
Chen, Tung-Sheng
學位類別: 碩士
Master
系所名稱: 生技醫藥產業碩士學位學程
Graduate Program of Biotechnology and Pharmaceutical Industries
論文出版年: 2020
畢業學年度: 109
語文別: 中文
論文頁數: 90
中文關鍵詞: 杜鵑花酸幹細胞皮膚老化微小分子核糖核酸
英文關鍵詞: Azelaic acid, Stem cells, Skin Aging, MicroRNA
DOI URL: http://doi.org/10.6345/NTNU202001712
論文種類: 學術論文
相關次數: 點閱:194下載:0
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  • 目錄 1 中文摘要 6 Abstracts 8 1.緒論 10 1-1皮膚 10 1-2皮膚光老化 11 1-3 microRNA(miRNA)調控老化機制 14 1-4目前皮膚抗老化的治療方式 16 1-5幹細胞用來改善皮膚老化的基本原理 17 1-6幹細胞種類 20 1-7幹細胞強化 20 1-8杜鵑花酸(Azelaic acid,AzA) 22 2.研究動機 25 3.研究材料與方法 26 3-1 化學藥品與試劑 26 3-2 人類角質細胞(Keratinocytes、HaCaT)培養 26 3-3 臍帶膠質間葉幹細胞(Wharton's Jelly Stem Cells,WJSC) 27 3-4 雙氧水(Hydrogen peroxide,H2O2)傷害HaCaT 27 3-5 AzA強化幹細胞 28 3-6 細胞存活率檢測(MTT assay) 28 3-7 蛋白質定量(細胞) 29 3-8 蛋白質定量(動物組織) 29 3-9 西方墨點法檢測(Western blot assay)(細胞) 30 3-10 西方墨點法檢測(Western blot assay) (動物組織) 31 3-11 小分子干擾核糖核酸(small interfering RNA,siRNA)轉染WJSC 33 3-12 miRNA轉染HaCaT 33 3-13 流式細胞儀分析(Flow Cytometry) 34 3-14 細胞分組 35 3-15 實驗動物 35 3-16 動物分組 35 3-17 皮膚老化動物模式的誘發 36 3-18 馬森三色染色法(Masson's trichrome stain) 36 3-19 病理組織化學染色(Hematoxylin and eosin stain、H&E stain) 37 3-20 病理組織免疫染色(Immunohistochemistry, IHC) 37 3-21 統計分析 38 4.實驗結果 39 4-1用H2O2誘導 HaCaT細胞老化細胞存活率檢測 39 4-2用H2O2誘導 HaCaT細胞老化西方墨點法檢測 39 4-3 miRDB網站搜尋miRNA 40 4-4 miRNA-7850-5p 短暫轉殖後西方墨點法檢測 40 4-5用AzA處理WJSC細胞存活率檢測 40 4-6用AzA處理WJSC流式細胞儀分析 41 4-7用AzA處理WJSC後轉染siCXCR4細胞存活率檢測 41 4-8用AzA處理WJSC後轉染siCXCR4西方墨點法檢測 42 4-9用AzA處理WJSC的條件培養液救援老化的HaCaT細胞存活率檢測 43 4-10用AzA和siCXCR4處理WJSC的條件培養液救援老化的HaCaT西方墨點法檢測 43 4-11 UVB誘導小鼠皮膚老化後用AzA處理 WJSC的條件培養液救援老化小鼠皮膚實驗設計 44 4-12 UVB誘導小鼠皮膚老化後用AzA處理WJSC的條件培養液救援老化小鼠皮膚外觀改善情況 45 4-13 UVB誘導小鼠皮膚老化後用AzA處理WJSC的條件培養液救援老化小鼠皮膚毛囊數目改善情況 45 4-14 UVB誘導小鼠皮膚老化後用AzA處理WJSC的條件培養液救援老化小鼠皮膚表皮層增厚情況 46 4-15 UVB誘導小鼠皮膚老化後用AzA處理WJSC的條件培養液救援老化小鼠皮膚真皮層的膠原蛋白流失情況 46 4-16 UVB誘導小鼠皮膚老化後用AzA處理WJSC的條件培養液救援老化小鼠皮膚膠原蛋白標記Collagen 1 A1表現程度 47 4-17 UVB誘導小鼠皮膚老化後用AzA處理WJSC的條件培養液救援老化小鼠皮膚老化標記p53表現程度 48 4-18 UVB誘導小鼠皮膚老化後用AzA處理WJSC的條件培養液救援老化小鼠皮膚老化標記p21表現程度 49 5.討論 50 6.結論 53 7.未來工作 54 7-1 AzA-WJSC-CM的可專利性 54 7-2是否能將AzA-WJSC-CM製成商業配方 54 7-3除了治療衰老,AzA-WJSC-CM是否可以治愈其他皮膚疾病 55 7-4探索幹細胞旁分泌蛋白途徑。 55 8.參考文獻 56 9.圖與表格 62

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