臺灣博碩士論文加值系統

背景：
皮膚為人體最大器官，其功能為人體對外界環境的保護屏障、防止人體過度流失水分等。在需多相關文獻指出毛蕊花苷具有抗發炎、抗氧化之效果，但對於皮膚上皮傷口癒合則無相關研究；另外也觀察毛蕊花苷對於毛髮生長之作用。台灣特有的牛樟芝作為中藥藥材具有許多功效，其中也具有抗發炎能力，因此進一步探討牛樟芝純化物SY-1 對於傷口癒合相關機制影響。
材料與方法：
以人類真皮層細胞Human dermal fibroblast (HDF) cell為研究模式。觀察毛蕊花苷(Acteoside)及牛樟芝萃取物(SY-1)對於傷口癒合、抗發炎作用機制；以wound healing assay確認毛蕊花苷及SY-1對人類真皮層細胞的癒合作用；以西方墨點法觀察毛蕊花苷及SY-1對生長因子及發炎因子訊息路徑相關蛋白質之活化情況；並以動物實驗應證之。
結果： 研究發現毛蕊花苷及SY-1具有促進生長因子蛋白表現作用，同時也能夠降低發炎反應。分析生長因子及其上下游訊息路徑，發現皆有受到活化及增加蛋白表現增加的現象。另外利用TNF-α誘導HDF細胞處於發炎反應，發現毛蕊花苷及SY-1能夠顯著抑制IL-1β與IL-6，並且影響NFκB入核路徑。最後以動物實驗方式應證，發現毛蕊花苷及SY-1能有效促進傷口癒合以及毛髮生長作用。
結論：
本研究顯示毛蕊花苷及 SY-1 促進人類真皮層細胞傷口癒合作用與抗發炎能力，能有效增加生長因子表現及抑制發炎因子的表現。顯示毛蕊花苷及SY-1能透過改善人類真皮層細胞修復及重建的作用，恢復上皮屏障，並且具有開發成為治療炎症病人用藥的潛力。

Background:
Skin is the largest organ of human which functions to protect our body against external environment from infectious or toxic agents and excessive water loss. With the essential role of skin in protection of human body, it is critically important to maintain a healthy and intact skin.Previously, literatures documented that acteoside and the extract from Antrodia cinnamomea displays significant anti-oxidative and anti-inflammatory activites in skin cell models. Despite of the significant beneficiary effects, the effect and mechanism of these agents remain to be investigated. In an effort to explore the potential applications of these agents in skin protection, the effect and mechanism of acteoside and the extract from Antrodia cinnamomea in skin wound healing and hair growth were studied.
Materials and Methods:
The human dermal fibroblasts (HDF) and C57BL/6J mouse models were used for this study. The effects of acteoside and SY-1, a phenolic compound isolated from Antrodia cinnamomea, on anti-inflammatory, wound healing, and hair growth were investigated. The non-infectious and infectious models were performed to assessed the efficacious of these agents in wound healing. To investigated the mechanism of action of these compounds, western blot analysis were used to explore the expression of growth factors, inflammatory cytokines that are associated with skin inflammation and wound healing.
Results:
Our results indicated that acteoside and SY-1 displayed a significant anti-inflammatory activity in human HDF cells without detectable cytotoxicity. These compounds suppressed the level of TNFα-induced pro-inflammatory cytokines including IL6, IL1β, and NF-κB in HDF cells. In addition, the expression level of vascular endothelial growth factor (VEGF) and transforming growth factor β (TGFβ) were induced by these compounds. Furthermore, these compounds conferred a significant increase in wound healing and hair growth activity in both of the in vitro HDF cells and in vivo mouse models.
Conclusion:
This study demonstrated that acteoside and SY-1 display a significant skin protective effect through anti-inflammatory, increased wound healing, and hair growth promoting activities in human HDF cell and animal models. These findings provide the molecular basis for the skin protective activity and potential dermal applications of acteoside and SY-1.

誌謝1
目錄2
圖目錄4
縮寫表7
中文摘要8
Abstract10
第一章 緒言12
第二章 材料與方法27
壹、 主要儀器及藥品試劑27
一、儀器及器材27
二、藥品試劑28
三、抗體29
四、Acteoside及SY-1來源29
貳、 方法30
一、Human dermal Fibroblast (HDF)細胞培養(Cell culture)30
二、蛋白質濃度測定（Protein assay）33
三、西方墨點法（Western blotting）34
四、細胞增生分析（Cell proliferation assay）41
五、刮傷癒合試驗（Wound healing assay）42
六、傷口癒合動物模式43
七、金黃色葡萄球菌之培養與計算44
八、抑菌環試驗46
九、傷口組織細菌含量試驗46
十、皮膚組織切片47
十一、小鼠皮膚組織萃取48
十二、毛髮生長動物模式49
十三、Masson trichrome stain50
第三章 結果 51
壹、 Acteoside及SY-1對Human dermal fibroblast(HDF)細胞毒性測試51
貳、 Acteoside及SY-1對Human dermal fibroblast(HDF)細胞migration之影響51
參、 Acteoside及SY-1對Human dermal fibroblast(HDF)生長機制之影響53
肆、 Acteoside及SY-1對Human dermal fibroblast(HDF)發炎機制之影響54
伍、 Acteoside及SY-1在非感染性創傷中，促進小鼠傷口癒合作用之影響55
陸、 Acteoside及SY-1在感染性創傷中，促進小鼠傷口癒合作用之影響57
柒、 Acteoside及SY-1促進小鼠毛髮生長作用之影響59
第四章 討論 60
第五章 結論 62
參考文獻 85

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