臺灣博碩士論文加值系統

紫外線UV-B隨著臭氧層的逐年減少而對人體造成的傷害逐年增加，除了會造成皮膚發炎紅腫之外也具致癌性。牛樟芝 ( Taiwanofungus camphoratus )，其子實體具有許多生理活性成分，在許多實驗中已證實牛樟芝子實體萃取物具有抗氧化的能力，因此本論文主要以探索能降低UV-B傷害的特定牛樟芝分離萃取物，並進一步闡明其對受損細胞的保護機制。
牛樟芝先以酒精粗萃取後再以管柱層析的方式分離成FK1、FK2、FK3、FK4、FK5、FK6、Me等七個劃分( fractions ) 。將上述七個劃分進行毒性測試後，發現只有Me fraction具有明顯促進纖維母細胞( CCD-966SK )存活率的效用。繼續在細胞修復試驗中先以UV-B ( 60 mj/cm2 )刺激後再加入Me ( 7.5 g/ml ) ， 發現CCD-966SK存活率分別於24小時及48小時 回復14 % 與18.9 % ，及遷移能力的顯著回復，並減少細胞凋亡10.6 % 與 減少DNA片斷化 20 % 。探討細胞修復機制後發現加入Me的組別p53、Bax、C-PARP表現量下降，p21、Pro-Casepase 3表現量回復，並使細胞週期停滯於G2/M期，抑制細胞走向凋亡並進行DNA修復。在動物試驗中先以UV-B ( 360 mj/cm2 )刺激表皮後，再以Me塗抹於表皮後發現Me加速了小鼠皮膚損傷修復的速度。本論文首次發現牛樟芝子實體之部份純化物Me具有降低UV-B誘導損傷的效用，未來應有機會發展為修復受損皮膚的新藥。

With the gradual thining of the earth’s ozone layer, the damage UVB light reaching to the ground more and more. Too much ultraviolet B exposure may cause skin irritation, swelling, and the worst will lead to cause skin cancer. Taiwanofungus camphoratus (TC) is a unique fungal species of Taiwan. Recently, many studies have indicated that the antioxidative effects of crude extracts of the TC fruiting body (TCFB) in different test models. The main aim of this stdy is to explore the specific extract component from TCFB to reduce UVB damage and further clarify its protective mechanisms against damaged cells.
By using column chromatography technique, the ethanol crude extracts of TCFB could be separated into seven characteristic fractions, named as FK1, FK2, FK3, FK4, FK5, FK6 and Me. After cell viability test we found Me fraction showed the best result in the proliferation of CCD-966SK fibroblast cell. In the cell repair tests, CCD-966SK cells prior irradiated with UVB ( 60 mj/cm2 ) before Me ( 7.5 g/ml ) treated, the result showed Me could significantly enhance the cell migration and recover cell viability up to 14 % and 18.9 % in 24 and 48 hours respectively. In cell cycle test, the result showed Me component reduced SubG1 phase 10.6%. In TUNEL assay, Me component reduced DNA fragmentation 20%. In Western blot, Me component reduced p53, Bax, C-PARP but recovered p21 and pro-casepase3 expression, induced cell cycle arrested at G2/M phase. These results could be interpreted as Me component could inhibit apoptosis of damaged cells by UVB and improved the DNA repair. In animal model experiment, prior to stimulate the skin with UVB ( 360 mj/cm2 ) and then applied the Me component on the damaged skin, the results showed Me could accelerate the speed of wound healing. This study first confirmed that Me component, partially purified from TCFB, could reduce UVB-induced damage. It should have the opportunity to develop into skin care agent in the future.

中文摘要 I
Abstract III
目錄 V
表目錄 VI
圖目錄 VII
第一章、前言 1
第二章、文獻回顧 3
第三章、研究目的 25
第四章、實驗材料與儀器 27
第五章、實驗方法 33
第六章、實驗結果 49
第七章、討論 85
第八章、結論 91
第九章、未來展望 93
第十章、參考文獻 95

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http//content .edu.tw/senior/chemistry/tp_sc/surround/ozone/right11.htm 引用日期 2013/6/19
http://en.wikipedia.org/wiki/Pyrimidine_dimer
引用日期 2013/6/19