淨白成分抑制酪胺酸酶活性和斑馬魚胚胎黑色素生成之研究

Study on the Whitening Components on Inhibition of Tyrosinase Activity and Melanin Production in Zebrafish Embryos

劉祖寧(Tsu-Ning Liou) ； 洪浚祥(Jun-Xiang Hong) ； 陳詠善(Yung-Shan Chen) ； 陳淑德(Su-Der Chen)

《宜蘭大學生物資源學刊》 16卷 (2020/12) Pp. 49-62

https://doi.org/10.6175/job.202012_16.0004

摘要

麴酸和熊果素是衛福部核准的美白成分，而白藜蘆醇亦有淨白效果，故本研究選用麴酸、熊果素及白藜蘆醇作為酪胺酸酶的抑制劑，探討其酵素抑制動力學機制，由於台灣已禁止化妝品進行動物實驗，故本研究改以利用斑馬魚胚胎作為淨白研究的平台。結果顯示在體外試驗中，100μg/mL的麴酸、熊果素及白藜蘆醇對酪胺酸酶的抑制率分別為77.82%、8.72%及59.85%。在酵素抑制動力學的部分，麴酸及熊果素均為競爭型抑制劑，而白藜蘆醇則為無競爭型抑制。進一步做動物試驗，將三種抑制劑添加於斑馬魚胚胎水溶液中，並於72小時後分析斑馬魚胚胎黑色素生成量，可以發現在100μg/mL的濃度下，與控制組相比，浸泡於麴酸、熊果素與白藜蘆醇相對黑色素含量分別降為65.24、53.42與67.38%，尤其以熊果素淨白效果最佳，和體外抑制酪胺酸酶的效果有很大的差異，故未來可使用斑馬魚胚胎作為淨白成分之篩選平台。

關鍵字

麴酸 ； 熊果素 ； 白藜蘆醇 ； 酪胺酸酶 ； 酵素抑制動力學 ； 斑馬魚 ； 黑色素

並列摘要

Kojic acid and arbutin are whitening ingredients approved by the Ministry of Health and Welfare, and resveratrol has a whitening effect. Here, koji acid, arbutin, and resveratrol were selected as model tyrosinase inhibitors to study their enzyme inhibition kinetic mechanism in this study. Since Taiwan has banned animal experiments on cosmetics, this study changed to use zebrafish embryos as an in vivo platform for whitening study. The results showed that 100 μg/mL kojic acid, arbutin, and resveratrol inhibited tyrosinase by 77.82%, 8.72%, and 59.85% in the in vitro test, respectively. Regarding enzyme inhibition kinetics, both kojic acid and arbutin were competitive inhibitors, while resveratrol seemed to be a non-competitive inhibitor. Follow-up animal experiments, inhibitors at 100 μg/mL were immersed in zebrafish embryos, and the amounts of melanin production in zebrafish embryos were analyzed 72 hours later. Compared to the control, relative contents of melanin in kojic acid, arbutin, and resveratrol decreased to 65.24, 53.42, and 67.38%, respectively. Arbutin showed the most promising whitening effect, and the effect of inhibiting tyrosinase in vitro was different. Therefore, using zebrafish embryos can be a feasible screening platform for detailed testing of whitening ingredients.

並列關鍵字

kojic acid ； arbutin ； resveratrol ； tyrosinase ； enzyme inhibition kinetics ； zebrafish ； melanin