臺灣博碩士論文加值系統

青春痘是一類常見的皮膚炎性疾病，發病的因素主要有不正常的角質化、皮脂過度的分泌及大量增生的痤瘡丙酸桿菌所造成的發炎反應等，本實驗為探討青春痘所介導的發炎路徑，以小鼠試驗建立青春痘發炎模式，並研究角質細胞表現接受器表現量以及在發炎中所扮演的趨化角色和痤瘡丙酸桿菌是如何影響異常角質化的相互作用。
經小鼠試驗結果顯示，當痤瘡丙酸桿菌注射至小鼠皮內後，在第7天檢測到代表TH1的IFN-γ開始表現且約維持7天，而代表TH2的IL-4、IL-5則沒有被檢測到，推測青春痘發炎路徑與TH1有關。在角質細胞接受器實驗中，經痤瘡丙酸桿菌感染誘導後TLR2表現量約1.3~1.8倍，推測角質細胞不是免疫細胞，接受器表現能力有限。而角質細胞異常角質化實驗以分化標記物角質素的變化進行探討，結果顯示當角質細胞經痤瘡丙酸桿菌感染時，代表角質細胞開始分化到棘狀層的第1、10、16型角質素會明顯提升，而位於基底層代表角質細胞具正常分裂、增殖功能的第5、14角質素上升幅度不明顯，推測痤瘡丙酸桿菌具有誘導分化能力。再進一步使用IFN-γ與痤瘡丙酸桿菌刺激角質細胞，探討角質細胞所釋放的趨化因子，兩次結果比較一致的有CXCL16、IL-8、MCP-1趨化因子在四個組別皆被檢測到，在第一次實驗中CXCL16、IL-8趨化因子在痤瘡丙酸桿菌組別升高，但第二次實驗卻沒有升高，MCP-1趨化因子則在兩次實驗中IFN-γ組別皆有升高。而在兩次實驗中最為一致的是I-TAC、IP-10趨化因子，在IFN-γ組別以及菌種與IFN-γ相加組別被檢測到，且在第一次實驗中具有加乘作用，未來可將較顯著差異的趨化因子進行單一分析，確認經由痤瘡丙酸桿菌所介導而表現趨化因子，進一步確認青春痘中可能表現趨化因子。

Acne is a common type of skin inflammatory disease. The incidence of the main factors abnormal keratinization, inflammation excessive secretion of sebum and Propionibacterium acnes caused. By this experiment is to explore mediated inflammatory acne pathway, in tests in mice models to establish inflammation of acne, and studies show keratinocyte receptor expression in inflammation and in the role played by chemokines and P. acnes is how it affects the interaction of abnormal keratinization.
The test results in mice show that when P. acnes injected into the skin of mice after the 7 days of detected representatives TH1 of IFN-γ began to expression and last for about 7 days, and on behalf of TH2 of IL-4, IL- 5 has not been detected, suggesting that inflammatory acne path TH1 related. In keratinocytes receptor experiments by P. acnes infection induced Toll-like receptor 2 expression is about 1.3 to 1.8 fold, speculate keratinocytes not that immune cells limited receptor performance capabilities. The abnormal keratinocyte keratinocytes differentiation markers change experiment keratin was investigated. The results show when keratinocytes over time P. acnes infection, keratinocytes begin to differentiation on behalf of the first 1,10,16 type keratan spinous layer It will be improved significantly, and is located in the basal layer of the representative of a normal keratinocytes division, section 5, 14 proliferation of keratin obvious rise, suggesting that P. acnes induce differentiation. Further use of IFN-γ and P. acnes stimulate the keratinocytes, that discuss the release of chemokines, the two results are more consistent CXCL16, IL-8, MCP-1 chemokine in all four categories is detected in the first experiment CXCL16, IL-8 chemokines in P. acnes groups increased, but the second test has not increased, MCP-1 chemokine in the two experiments IFN-γ groups increased. In both experiments was the most consistent I-TAC, IP-10 chemokine in the IFN-γ group and adding the bacteria with IFN-γ groups were detected, and having in the first experiment synergistic effect, the future may be the more significant differences chemokine single analysis, confirmed by P. acnes mediated expression of chemokines and further confirmation acne may show chemokines.

中文摘要 I
Abstract II
誌謝 IV
目錄 V
表目錄 VII
圖目錄 VIII
第一章、文獻回顧 1
1.1青春痘 1
1.2青春痘發炎路徑 3
1.3 青春痘相關接受器 6
1.4痤瘡丙酸桿菌 8
1.5角質細胞 9
1.6 實驗目的 13
第二章、材料與方法 14
2.1實驗材料 14
2.1.1細菌培養 14
2.1.2細胞培養 14
2.1.3 RNA萃取與聚合酶連鎖反應檢測 15
2.1.4 酵素免疫吸附試驗檢測 15
2.1.5 蛋白質芯片分析 15
2.2實驗方法 16
2.2.1細菌培養 16
2.2.2細胞培養 16
2.2.3細胞存活率試驗 17
2.2.4細胞、細菌共培養 18
2.2.5細胞RNA萃取 18
2.2.6反轉錄聚合酶連鎖反應 19
2.2.7聚合酶連鎖反應 19
2.2.8即時定量聚合酶連鎖反應 19
2.2.9蛋白質芯片分析 20
2.2.10實驗動物 21
2.2.11菌種製備 21
2.2.12注射與採樣 22
2.2.13酵素免疫吸附試驗 22
第三章、結果 23
3.1建立青春痘發炎小鼠模式 23
3.2細胞存活率試驗 26
3.3角質細胞經感染後表現接受器 28
3.4角質細胞經感染後角質素變化 30
3.5角質細胞經感染後表現趨化因子 35
第四章、討論 46
第五章、參考文獻 49

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