黑豆種皮乙醇萃取物對人類黑色素瘤細胞黑色素形成之抑制效果

In vitro anti-melanogenesis effect of crude extract from seed coat of black soybean toward human malignant melanoma A375.S2 cell line

詹鈞凱(Jyun-Kai Jhan)

指導教授 ： 許成光

亞洲大學/健康學院/生物科技學系碩士班/碩士(2009年)

摘要

本論文主要探討黑豆(台南三號)之種皮50％乙醇萃取物對酪胺酸酵素活性抑制影響，並利用人類黑色素瘤細胞A375.S2細胞株，進一步研究黑豆種皮50％乙醇萃取物對體外黑色素生成之抑制影響。於酪胺酸酵素活性抑制實驗之結果顯示，黑豆50％乙醇萃取物對酪胺酸酵素活性有抑制影響，於同濃度(1.66 mg/ml)下黑豆種皮50％乙醇萃取物對於酪胺酸酵素活性抑制率為黑豆整顆50％乙醇萃取物之兩倍以上，顯示大部份之抑制酪胺酸酵素有效成份存在於黑豆種皮中，且抑制率隨著黑豆種皮50％乙醇萃取物濃度提高而增加，抑制機制屬於混合型抑制作用。於A375.S2細胞培養試驗之結果顯示，A375.S2細胞株中含有酪胺酸酵素活性可與受質進行反應生成黑色素前驅物，於黑豆50％乙醇萃取物加藥處理後，以三種不同濃度之L-3,4-dihydroxy- phenylalanine (L-dopa) 0.05, 0.25 and 0.5 mg/ml作為受質，最高的兩個黑豆50％乙醇萃取物濃度(20 μg/ml與30 μg/ml)對於A375.S2細胞株中酪胺酸酵素活性有較明顯的抑制效果。

關鍵字

黑豆 ； 酪胺酸酵素 ； 多巴胺 ； 人類黑色素瘤細胞株

並列摘要

The purposes of this study were to evaluate the inhibitory potential of 500 mg/L ethanol extract from the seed coat of black soybean (Glycine max (L.) Merrill, Tainan No. 3) (BSSCEE) toward the activity of tyrosinase, otherwise human malignant melanoma A375.S2 cell line was used to assess the in vitro effect of BSSCEE on the melanogenesis. The results of inhibitory effects on mushroom tyrosinase activity showed that BSSCEE could inhibit the activity of mushroom tyrosinase. At the same concentration (1.66 mg/ml) between BSSCEE and 500 mg/L ethanol extract from the whole black soybean (WSSEE), the inhibition rate of BSSCEE better than WSSEE exceed in double. Indicate that the most of inhibitors on mushroom tyrosinase were existed in the seed coat of black soybean. Otherwise the inhibition rate was depend on the concentration of BSSCEE and the inhibition mechanism was belong to mixed-type inhibition. The results of cell culture were showed that there was the tyrosinase ability to catalyze L-dopa during human malignant melanoma A375.S2 cell line. After treated with different concentrations of BSSCEE, using three different concentrations of L-dopa (0.05, 0.25 and 0.5 mg/ml) to evaluate. Among all of results the highest two concentrations of BSSCEE (20 μg/ml and 30 μg/ml) showed the more significant inhibitory effect on human malignant melanoma A375.S2 cell line.

並列關鍵字

black soybean ； tyrosinase ； L-3,4-dihydroxyphenylalanine ； A375.S2 cell

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