A new mechanism of action for skin whitening agents: binding to the peroxisome proliferator-activated receptor

Synopsis Octadecenedioic acid is known as a skin whitening agent but its activity is not mediated via a direct inhibition of tyrosinase. Based on the secondary properties of this molecule, such as its anti-inflammatory and anti-ageing effects, we postulated that octadecenedioic acid interacted with the peroxisome proliferator-activated receptor (PPAR) as this nuclear receptor also mediates these effects. Using reporter gene technology, we were indeed able to demonstrate binding of octadecenedioic acid to all three PPAR subtypes, in particular PPARgamma with an EC(50)-value of approx. 1 x 10(-6) m. Binding to PPARgamma of octadecenedioic acid or rosiglitazone, a known pharmaceutical PPARgamma agonist, led to reduced melanogenesis. Subsequently also tyrosinase mRNA (as measured by real-time polymerase chain reaction) and tyrosinase levels (as measured by Western blot) were reduced, suggesting the existence of a complete novel mechanism of skin whitening agents: binding to PPARgamma results in reduced tyrosinase mRNA expression which in turn results in less tyrosinase being formed. This in turn leads to reduced melanogenesis both in vitro and in vivo Because octadecenedioic acid binds not only to PPARgamma but also to PPARalpha and PPARdelta, other efficacies mediated via these receptors may also be expected.     

植物中酪氨酸酶的筛选及不同抑制剂对其活性的影响

以新鲜马铃薯、红富士苹果、红蛇果、秀珍菇和紫茄子为研究对象,从中分别提取酪氨酸酶并测定其酶活力,发现紫茄子中提取的酪氨酸酶具有最大酶活力。因此选紫茄子中提取的酪氨酸酶做进一步研究。选邻苯二酚作为茄子酪氨酸酶底物,进一步研究了有机类、无机类和天然类三大类抑制剂对酪氨酸酶催化活性的影响,并测定了这些抑制剂的IC50值。     

曲酸与阿魏酸对酪氨酸酶的抑制作用研究

本实验采用Lineweaver-Burk双倒数法探讨了曲酸与阿魏酸对蘑菇酪氨酸酶催化L-DOPA氧化的抑制作用并推测其抑制机理。通过对曲酸与阿魏酸的IC50与抑制常数KI的测定可知,曲酸对酪氨酸酶的抑制作用明显高于阿魏酸。分别采用分光光度法和ImageJ图像分析与处理软件定量地测定曲酸与阿魏酸对虾血淋巴和苹果褐变的抑制作用。曲酸对虾血淋巴褐变的抑制作用优于阿魏酸;对于苹果褐变,阿魏酸的抑制作用则高于曲酸。     

Artocarpus lakoocha heartwood extract as a novel cosmetic ingredient: evaluation of the in vitro anti-tyrosinase and in vivo skin whitening activities

The heartwood extract of Artocarpus lakoocha Roxb. was evaluated for the in vitro tyrosinase inhibitory activity and the in vivo melanin-reducing efficacy in human volunteers. The IC(50) of the extract and oxyresveratrol, its major active ingredient, against mushroom tyrosinase was determined to be 0.76 and 0.83 mug mL(-1), respectively. The extract dissolved in propylene glycol was subsequently tested in female volunteers using a parallel clinical trial with self-control (n = 20 per group). The first group received the 0.25% w/v A. lakoocha solution as the test solution, whereas the second and the third group, respectively, received 0.25% licorice extract and 3% kojic acid as the reference solutions in the same solvent. The subjects in each group twice daily applied the test (or reference) solution in one of her upper arm, whereas the remaining arm was treated with only propylene glycol (self-control) for 12 weeks. The melanin content of each application site was measured using Mexameter every week and calculated as % reduction in melanin content relative to the initial melanin value (% whitening). The value of % whitening was then compared between the product-treated and the propylene glycol-treated arms within the same subject using paired t-test (alpha = 0.05). The A. lakoocha extract was the most effective agent, giving the shortest onset of significant whitening effect after only 4 weeks of application (P < 0.05), followed by 3% kojic acid (6 weeks) and 0.25% licorice extract (10 weeks). The effect also increased with time with maximum whitening observed at week 12 for A. lakoocha extract. When the extract was formulated as an oil-in-water emulsion, its whitening efficacy was further enhanced. Daily application of 0.1% w/w A. lakoocha lotion to the upper arms (n = 25) and cheeks (n = 15) of volunteers produced significant whitening over the lotion base after 2 and 3 weeks, respectively (P < 0.05). Thus, the preliminary study suggested that the heartwood extract of A. lakoocha may have a promising potential for use as an effective and economical skin-whitening agent.     

白藜芦醇对酪氨酸酶体外抑制活性的动力学研究

在30℃,pH 6.8的Na2HPO4-NaH2PO4的缓冲体系中,采用酶促动力学方法,研究了白藜芦醇对酪氨酸酶单酚酶和二酚酶的抑制作用。结果表明,白藜芦醇对酪氨酸酶、单酚酶和二酚酶均有抑制作用,对单酚酶抑制活性的IC50值(抑制率达到50%时的白藜芦醇质量浓度)约为5.1mg/mL,对二酚酶抑制活性的IC50值约为5.6 mg/mL。此外,白藜芦醇可延长单酚酶的迟滞效应,8 mg/mL的白藜芦醇能使迟滞时间从22 s延长至62 s,而对二酚酶则无此迟滞作用。Lineweav-er-Burk图分析表明,白藜芦醇对酪氨酸酶的抑制作用为混合型抑制,对游离酶的抑制常数(KI)和对酶-底物络合物的抑制常数(KIS)分别为3.4 mg/mL和35.98 mg/mL。     

青梅花提取物的酪氨酸酶抑制作用及机理研究

目的：以青梅花为原料,探讨其提取物（PF）在体外对酪氨酸酶的抑制作用及机理。方法：首先采用多巴速率氧化法体外测定PF酪氨酸酶的抑制活性,并绘制Lineweaver-Burk曲线,推断其抑制类型。分别采用化学发光法和铁氰化钾还原法测定PF的清除自由基能力及还原力。结果表明：PF能显著抑制酪氨酸酶活性,其半抑制浓度IC50为（98．89±6．34）ug／mL,抑制类型为混合型抑制。PF对酪氨酸酶的抑制作用可能与提取物中绿原酸等多酚物质、较强的自由基清除能力和还原力有关。结论：PF是一种有前途的酪氨酸酶抑制剂,可应用于果蔬等食品的加工和贮藏。     

沙棘黄酮对酪氨酸酶活力抑制作用的研究

研究沙棘黄酮对酪氨酸酶活性的抑制作用。实验测定了沙棘黄酮分别在温度、pH、时间、底物浓度、酶液加入量等因素条件下对酪氨酸酶活力的影响。结果显示,沙棘黄酮对酪氨酸酶活性的抑制率随着时间的延长,底物浓度的增加,酶量的增加而增强,而且在35℃及pH7.1时沙棘黄酮对酪氨酸酶活力的抑制作用最强可达到47.45%。说明沙棘黄酮对酪氨酸酶的活性具有较强的抑制作用。     

白附子酪氨酸酶活性抑制成分的提取与分离

研究了不同溶剂及提取方式对白附子中酪氨酸酶抑制成分的影响,并应用活性追踪法对白附子中酪氨酸酶活性抑制成分进行考察。结果表明,以20%乙醇水溶液为溶剂从白附子中提取的粗提物对酪氨酸酶活性的抑制作用最强,抑制率可达到96.35%,与恒温水浴振荡提取法相比,超声波辅助提取法对活性成分的提取效果更好。在此提取条件下所得白附子粗提物对酪氨酸酶的半抑制浓度IC50为24.69mg/mL。该粗提物依次经乙醚和正丁醇萃取,所得正丁醇萃取组分对酪氨酸酶的抑制率达71.94%。对高活性的正丁醇萃取组分进行分离,获得了一种酪氨酸酶抑制率高达98.82%的组分,是一种潜在的酪氨酸酶抑制剂。研究结果对白附子及其活性组分在医药和食品领域的应用有指导意义。     

发酵法制曲酸的研究应用

本文概述了曲酸特有的分子结构及产品性质,同时介绍了其在化妆品、食品、医药、生物农药等领域的良好用途。说明曲酸尽快实现工业化生产在我国已迫在眉睫。