| 摘要: | 麴酸為目前衛生署訂定之美白成分，主要從真菌類Aspergillus與Penicillium之醱酵液中獲得，其作用機轉為抑制酪胺酸酶的活性，藉此達到防止黑色素生成之幼纂C然而高劑量麴酸會造成人體致癌與基因毒性等變異性風險。本研究目的擬由米麴菌醱酵後之產物尋找具應用潛力之美白原料並探討其作用機轉。
本研究利用中草藥（薑、黃耆、白芍、黃芩、麥門冬）與異黃酮指標成分Daidzein、Genistein經由米麴菌(Aspergillus oryzae)醱酵後，與中草藥醱酵產物5784’-TF (5,7,8,4'-Tetrahydroxyisoflavone)、784'-TF (7,8,4'-Trihydroxyisoflavone)探討具有抑制B16小鼠黑色素瘤細胞內酪胺酸酶活性與減少黑色素生成作用。利用體外模式系統評估中草藥經米麴菌醱酵產物與5784'-TF對蘑菇酪胺酸酶活性抑制之影響。結果顯示，濃度0.1mg/mL之黃耆未經由米麴菌醱酵產物（黃耆-前）與薑、麥門冬經由米麴菌醱酵後代謝產物（薑-後、麥門冬-後），皆有抑制蘑菇酪胺酸酶之活性，抑制率分別為51.3%、37.6%、31.4%；於濃度0.1mM daidzein、genistein及其醱酵產物5784'-TF抑制率分別為61.4%、63.4%、66.9%。另一方面，建立B16小鼠黑色素瘤細胞測試平台，其結果顯示薑、黃耆經由米麴菌醱酵後之代謝產物均可減少黑色素含量，與抑制細胞內酪胺酸酶活性且呈現劑量依存反應(dose-dependent reaction)。而白芍、黃芩、麥門冬之醱酵後產物則須在0.1mg/mL濃度下，才有顯著抑制效果。然而daidzein、genistein經米麴菌醱酵後之代謝產物784'-TF、5784'-TF比醱酵前均可抑制黑色素瘤細胞胞外釋放黑色素含量，亦可減少胞內黑色素生成，其中皆可抑制催化黑色素生成之酵素活性以降低黑色素細胞內黑色素生合成。此外，以tyrosinase zymography試驗，再次確認薑、黃耆及其醱酵產物確實能夠抑制細胞內酪胺酸酶活性之表現。再以CIE L*a*b*皮膚色澤計、多弁鄍祧尾侅t統，進行受測樣品實際應用於人體皮膚之美白有效性評估試驗，其結果發現，十位受測者塗抹乳液後皆具顯著改善膚色、降低黑色素生成之效果。
經由上述結果證實，薑、黃耆與異黃酮指標成分經由米麴菌(Aspergillus oryzae )醱酵後之代謝產物784'-TF、5784'-TF，較未經由米麴菌醱酵之成分具有抑制酪胺酸酶活性與減少黑色素生成作用。本研究推測經由米麴菌醱酵後之代謝產物，釋放酵素將原存中草藥化學分子進行生物轉化作用並催化形成更具美白之有效成分。因此，期望未來可應用於美白化粧品新成份開發之參考依據。
Kojic acid (5-hydoxy-4-pyran-4-one-2-methyl) is a tyrosinase inhibitor that it derived from various fungal species such as Aspergillus and Penicillium. However, some reports showed the carcinogenic potential and genotoxic risk of kojic acid.
This study hyperpigmentation of the mechanism has attempted to find effect of whitening agent fermented from Aspergillus oryzae.The inhibition of tyrosinase activities and melanin formation in B16 melanoma cells of fermented or nonfermented by Aspergillus oryzae from traditional Chinese medical herbs (Rhizoma Zingiberi,Radix Astragali, Paeoniae Albiflorae Radix, Scutellatiae Radix,Ophiopogonis Tuber) and the purified isoflavones from fermented 5784'-TF(5,7,8,4'-Tetrahydroxyisoflavone),784'-TF(7,8,4'-Trihydroxyisoflavone ) were evaluated in this study.
The experiments were evaluation mushroom tyrosinase activities of the extracts of Chinese medical herbs (Radix Astragali, Rhizoma Zingiberi, Ophiopogonis Tuber) and purified isoflavones by Aspergillus oryzae from fermented 5784'-TF, In vitro. Chinese medical herbs show that at 0.1mg/mL concentration was higher inhibition tyrosinase activity with data of 51.3%, 37.6%, 31.4%, and Daidzein, Genistein , 5784'-TF at 0.1mM concentration decrease 61.4%, 63.4%, 66.9%.
The results show that Rhizoma Zingiberi, Radix Astragali fermented with Aspergillus oryzae significantly reduce the cellular melanin content and inhibitory tyrosinase activity in a dose-dependent reaction. While that at 0.1mg/mL concentraction, fermented Chinese medical herb extracts can cause different effect of inhibiting tyrosinase that Paeoniae Albiflorae Radix, Scutellatiae Radix, Ophiopogonis Tuber. All results show that 784'-TF, 5784'-TF can inhibit the melanin formation and secretion from B16 melanoma cells after fermentation treatment.
Tyrosinase zymography was use to make sure Rhizoma Zingiberi, Radix Astragali fermented from Aspergillus oryzae reduce B16 cellular tyrosinase activity expression. In this study, we use a chromatography with CIE L*a*b* colour system to observe the whitening efficacy when apply on the human skin. The results show that ten voluteers all apparently improving, then confirmed the significantly reduce melanogenesis after the cream treatment.
Our conclusion show that Chinese medical herbs (Rhizoma Zingiberi, Radix Astragali, Paeoniae Albiflorae Radix, Scutellatiae Radix, Ophiopogonis Tuber) and purified isoflavones by Aspergillus oryzae from fermented products, significantly reduce the cellular melanin and inhibit tyrosinase activity in B16 melanoma cell.This study guess the biotransformation of secretion with Chinese medical herbs by enzymes and catalyzed as effective skin-whitening angents from fermented with Aspergillus oryzae.Therefore,there will developed effective skin-whitening angents for cosmetic markets in future. |
