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研究生:謝宛諭
研究生(外文):Wan-Yu Hsieh
論文名稱:玉蘭花萃取物美白活性及抗氧化能力之研究
論文名稱(外文):Inhibition of melanogenesis and antioxidant properties of Magnolia grandiflora L. flower extract
指導教授:張聰民張聰民引用關係
指導教授(外文):Tsong-Min Chang
學位類別:碩士
校院名稱:弘光科技大學
系所名稱:化妝品科技研究所
學門:民生學門
學類:美容學類
論文種類:學術論文
畢業學年度:100
語文別:中文
論文頁數:69
中文關鍵詞:玉蘭花黑色素生成酪胺酸酶黑色素抗氧化
外文關鍵詞:Magnolia grandiflora Lmelanogenesistyrosinasemelaninantioxidant
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玉蘭花是一種極具台灣特色之花卉,廣泛使用在亞洲作為傳統草藥藥物。以往的研究報告指出,玉蘭花葉之乙醇萃取物具有抗真菌、抗胃潰瘍和抗氧化活性等,針對其花瓣之萃取物的美白和抗氧化能力研究則未曾被探討。本研究利用分光光度計來測定玉蘭花萃取物對於蕈菇酪胺酸酶的活性,在B16F10小鼠黑色素瘤細胞中酪胺酸酶的活性和黑色素含量的抑制作用;利用西方墨點法來測定黑色素生成路徑相關的蛋白質表達。同時還探討了玉蘭花萃取物對於清除自由基能力及抗氧化活性。結果顯示,玉蘭花萃取物可降低蕈菇酪胺酸酶活性(IC50= 11.1 %),在B16F10小鼠黑色素瘤細胞中有效降低黑色素含量(IC50= 25.6 %)和抑制細胞內酪胺酸酶活性(IC50= 13.6 %),上述生物活性皆呈現劑量依賴性,玉蘭花萃取物也明顯減少Tyrosinase和TRP-1兩種蛋白的表現。在抗氧化方面,總酚含量測定的結果是最好的,其次是ABTS+自由基清除和還原力。我們得到的結論是玉蘭花萃取物可能是經由抑制Tyrosinase和TRP-1的表現或是降低活性氧(ROS)的含量,達到降低細胞中黑色素含量的效果,亦具有抗氧化能力。因此玉蘭花萃取物可以作為一種皮膚美白劑應用添加於保養品中。
Magnolia grandiflora L. flower is a very special plant species in Taiwan, also widely used as traditional herbal medicines in Asia. Previous studies have demonstrated that Magnolia grandiflora L. ethanol extract has anti-fungal, anti-ulcer and antioxidant activities. However, the whitening and antioxidant capacities of Magnolia grandiflora L. have not been explored. In this study, the effects of Magnolia grandiflora L. aqueous extract on mushroom tyrosinase activity, B16F10 murine melanoma cellular tyrosinase activity and cellular melanin content were determined spectrophotometrically. The expression of tyrosinase and related proteins in B16F10 cells were determined by Western blotting. Meanwhile, the antioxidative capacities of the flower extract were also investigated. Our data indicate that Magnolia grandiflora L. flower extract reduce mushroom tyrosinase activity (IC50= 11.1%), the flower extract also decrease the amount of melanin (IC50= 25.6%) and effectively suppressed intracellular tyrosinase activity (IC50= 13.6%) in a dose-dependent manner in B16F10 cells. Protein expression level of tyrosinase and tyrosinase-related protein 1 (TRP-1) were also decreased by the flower extract. Additionally, The extract exhibited apparent total phenolic content, ABTS+ free radical scavenging activity and reducing capacity. Our results conclude that Magnolia grandiflora L. flower extract inhibited melanogenesis in B16F10 cells through decreased the expression of tyrosinase and TRP-1 or depletion of cellular reactive oxygen species (ROS).These findings indicate that Magnolia. grandiflora L. could be applied as a type of skin-whitening agent in skin care products.
誌謝 I
中文摘要 II
英文摘要 III
目錄 IV
圖目錄 VII
縮寫表 VIII
壹、緒論 1
一、玉蘭花 1
(一)玉蘭花的介紹 1
(二)玉蘭花的成分組成 1
(三)玉蘭花的生物活性 3
二、黑色素生合成 4
(一)黑色素 4
(二)酪胺酸酶 8
(三)相關蛋白表現機制 9
(四)訊息調節劑 13
三、抗氧化 14
(一)自由基 14
(二)活性氧 15
(三)自由基的產生 15
(四)自由基對生物體分子的傷害 15
(五)抗氧化劑 18
貳、研究目的與實驗架構 20
一、研究目的 20
二、實驗架構 21
參、材料與方法 22
一、實驗材料 22
(一)細胞株 22
(二)樣品來源 22
二、實驗方法 22
(一)細胞培養 22
(二)細胞存活率分析 23
(三)美白能力測定 24
(四)抗氧化能力測定 31
(五)統計分析 38
肆、實驗結果 39
一、美白能力測定 39
(一)玉蘭花萃取物對B16F10細胞存活率之影響 39
(二)玉蘭花萃取物對蕈菇酪胺酸酶活性之影響 39
(三)玉蘭花萃取物對B16F10細胞黑色素含量之影響 39
(四)玉蘭花萃取物對B16F10細胞酪胺酸酶活性之影響 39
(五)訊息調節劑對玉蘭花萃取物影響B16F10細胞黑色素含量之改變 40
(六)玉蘭花萃取物對B16F10細胞內蛋白質表現量之影響 40
二、抗氧化能力測定 41
(一)玉蘭花萃取物對於DPPH自由基清除能力測定 41
(二)玉蘭花萃取物之總酚含量測定 41
(三)玉蘭花萃取物之ABTS+自由基清除能力測定 41
(四)玉蘭花萃取物之還原力測定 41
(五)玉蘭花萃取物之螯合金屬離子能力測定 42
(六)玉蘭花萃取物對B16F10細胞內ROS表現量之影響 42
(七)玉蘭花萃取物對B16F10細胞內RS表現量之影響 42
伍、討論 43
陸、結論 46
柒、參考文獻 46
捌、結果附圖 55
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