中草藥美白活性研究 五倍子對B16黑色素細胞黑色素形成之抑制研究

Whitening Activity of Chinese Herbal Medicines The inhibition of melanin biosynthesis of Galla Rhois on B16 cell

張瑋玲(Wei-Lin Chang)

指導教授 ： 施純明 ； 王靜瓊

臺北醫學大學/醫學院/醫學科學研究所/碩士(2005年)

摘要

現今草本精華之美白化妝品廣受女性朋友歡迎，所以本研究目的是對中草藥的美白活性成份研究。首先檢測以70 %丙酮或50 %乙醇萃取之30種中草藥，對抗氧化能力和黑色素生成的影響。結果顯示：五倍子、大黃、頷垂豆、安石榴、玫瑰具有較明顯之抑制酪胺酸?(Tyrosinase)之活性，其IC50 分別為 22.00、77.75、80.04、120.13及127.00 mg/ml。且其清除DPPH自由基之活性，IC50分別為為3.78、23.19、24.87、11.77及14.62 mg/ml，其中以五倍子效果最好。再以a-MSH、UVA刺激B16細胞黑色素形成之細胞模式，進行五倍子之黑色素形成抑制研究，結果顯示:五倍子在無誘導劑處理細胞、UV照射兩天及a-MSH合併誘導誘導之下，均呈劑量依存性之抑制作用。另以Folin-Ciocalteu reaction及HPLC分析其主要植物化學成份分佈：得知五倍子富含總多酚類，且其主成份為1,2,3,4,6-Penta-O- -galloyl-b-D-glucose (PG)，再利用相同實驗方法分析PG的酪胺酸?抑制作用，其IC50為 15 mM，且經由酵素動力學分析，屬於非競爭型抑制劑(Non-competitive inhibitor)。最後以親水軟膏為基劑，調製五倍子萃取物軟膏，進行動物皮膚刺激性試驗。觀察結果發現：2 g/g (w/w)、1 g/g (w/w)之五倍子濃度均不會引起小鼠皮膚過敏。所以，經由上述結果推論，五倍子應具皮膚美白化妝品開發研究之潛力。

關鍵字

B16黑色素細胞 ； 美白 ； 酪胺酸?

並列摘要

Nowadays herbal medicines of whitening cosmetic are popular for women. For this reason, we attempted to find the whiting activity compound present in many herbal medicines used. First screen 70 % acetone or 50 % ethanol extracts of 30 differents kinds of herbal medicines that effects of antioxidation and melanin biosynthesis. Results showed that: Rhus javanica Linne 、Rheum palmatum Linne、Archidendron lucidum Benth、Punica granatum Linne and Rosa rugosa Thunberg showed a potent tyrosinase activity inhibition , IC50 value was 22.00, 77.75, 80.04, 120.13 and 127.00 mg/ml. The antioxidation of DPPH, IC50 value was 3.78、23.19、24.87、11.77 and 14.62 mg/ml, Rhus javanica Linne showed the most potent effect. In the cell culture assay use a-MSH、UVA induce melanogenesis in B-16 mouse melanoma cell. The results of the inhibitory effect of Rhus javanica Linne with no inducer and with inducer a-MSH co-culture and radiation UVA 2 days on melanin biosynthesis was observed in B-16 mouse melanoma cell all in a dose dependence. Then use Folin-Ciocalteu reaction and HPLC assay was used to identify the major compound: Rhus javanica Linne was richly in polyphenol and the major component was 1,2,3,4,6-Penta-O- galloyl-b-D-glucose (PG), and use the same tyrosinase activity inhibition assay PG whose IC50 value was 15 mM. By mushroom tyrosinase inhibition kinetics assay, the inhibition mechanism obtained from Lineweaver-Burk plots showed that 1,2,3,4,6-Penta-O-galloyl-b-D- -glucose is a non-competitive inhibitor. Then skin stimulate tests in animal modesl, results showing that both 2 g/g (w/w) and 1 g/g (w/w) Rhus javanica Linne, was not induce an allergic reaction in mice skin. Overall, the results indicate that the Rhus javanica Linne could be development as whitening cosmetic for skin care.

並列關鍵字

B16 melanoma cell ； melanin biosynthesis ； Tyrosinase

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