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人参(Panax ginseng C.A. Meyer)乙酸乙酯提取物及其主要成分通过抑制B16小鼠黑色素瘤细胞中的氧化应激来抑制α-促黑素细胞激素诱导的黑色素生成。

Ethyl acetate extract from Panax ginseng C.A. Meyer and its main constituents inhibit α-melanocyte-stimulating hormone-induced melanogenesis by suppressing oxidative stress in B16 mouse melanoma cells.

作者信息

Jiang Rui, Xu Xiao-Hao, Wang Ke, Yang Xin-Zhao, Bi Ying-Fei, Yan Yao, Liu Jian-Zeng, Chen Xue-Nan, Wang Zhen-Zhong, Guo Xiao-Li, Zhao Da-Qing, Sun Li-Wei

机构信息

Jilin Technology Innovation Center for Chinese Medicine Biotechnology, College of Biology and Chemistry, Beihua University, Jilin City, Jilin Province 132013, China.

Changchun University of Chinese Medicine, Changchun, Jilin Province 130000, China.

出版信息

J Ethnopharmacol. 2017 Aug 17;208:149-156. doi: 10.1016/j.jep.2017.07.004. Epub 2017 Jul 8.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Hyperpigmentation disease involves darkening of the skin color due to melanin overproduction. Panax ginseng C.A. Meyer is a well-known traditional Chinese medicine and has a long history of use as a skin lightener to inhibit melanin formation in China, Korea and some other Asian countries. However, the constituents and the molecular mechanisms by which they affect melanogenesis are not fully clear.

AIM OF THE STUDY

The purpose of this study was to identify the active ingredient in Panax ginseng C.A. Meyer extract that inhibits mushroom tyrosinase activity and to investigate the antioxidative capacity and molecular mechanisms of the effective extract on melanogenesis in B16 mouse melanoma cells.

MATERIALS AND METHODS

Aqueous extracts of Panax ginseng C.A. Meyer were successively fractionated with an equal volume of chloroform, ethyl acetate, and n-butyl alcohol to determine the effects by examining the activity of mushroom tyrosinase. The effective fraction was analyzed using HPLC and LC-MS. The antioxidative capacity and the inhibitory effects on melanin content, cell intracellular tyrosinase activity, and melanogenesis protein levels were determined in α-melanocyte-stimulating hormone (α-MSH)-treated B16 mouse melanoma cells.

RESULTS

The ethyl acetate extract from Panax ginseng C.A. Meyer (PG-2) had the highest inhibiting effect on mushroom tyrosinase, mainly contained phenolic acids, including protocatechuic acid, vanillic acid, p-coumaric acid, salicylic acid, and caffeic acid, and exhibited apparent antioxidant activity in vitro. PG-2 and its main constituents significantly decreased melanin content, suppressed cellular tyrosinase activity, and reduced expression of tyrosinase protein to inhibit B16 cells melanogenesis induced by α-MSH, and no cytotoxic effects were observed. They also inhibited cellular reactive oxygen species (ROS) generation, increased superoxide dismutase (SOD) activity and glutathione (GSH) level in α-MSH-treated B16 cells effectively. And those activities of its main constituents could reach more than 80% of PG-2. The ROS scavengers N-acetyl-L-cysteine (NAC) had a similar inhibitory effect on melanogenesis.

CONCLUSIONS

These results suggest that ethyl acetate extract from Panax ginseng C.A. Meyer has the highest effect on inhibiting melanogenesis, and that its main components are polyphenolic compounds, which may inhibit melanogenesis by suppressing oxidative stress. This work provides new insight into the active constituents and molecular mechanisms underlying skin-lightening effect of Panax ginseng C.A. Meyer.

摘要

民族药理学相关性

色素沉着疾病是由于黑色素过度生成导致皮肤颜色加深。人参是一种著名的传统中药,在中国、韩国和其他一些亚洲国家,作为一种抑制黑色素形成的皮肤美白剂有着悠久的使用历史。然而,其影响黑色素生成的成分和分子机制尚不完全清楚。

研究目的

本研究旨在确定人参提取物中抑制蘑菇酪氨酸酶活性的活性成分,并研究有效提取物对B16小鼠黑色素瘤细胞黑色素生成的抗氧化能力和分子机制。

材料与方法

人参水提取物依次用等体积的氯仿、乙酸乙酯和正丁醇进行分级分离,通过检测蘑菇酪氨酸酶的活性来确定其效果。有效部位采用高效液相色谱(HPLC)和液相色谱-质谱联用(LC-MS)进行分析。在α-黑素细胞刺激素(α-MSH)处理的B16小鼠黑色素瘤细胞中测定抗氧化能力以及对黑色素含量、细胞内酪氨酸酶活性和黑色素生成蛋白水平的抑制作用。

结果

人参乙酸乙酯提取物(PG-2)对蘑菇酪氨酸酶的抑制作用最强,主要含有酚酸类成分,包括原儿茶酸、香草酸、对香豆酸、水杨酸和咖啡酸,并且在体外表现出明显的抗氧化活性。PG-2及其主要成分显著降低了黑色素含量,抑制了细胞酪氨酸酶活性,并降低了酪氨酸酶蛋白的表达,从而抑制α-MSH诱导的B16细胞黑色素生成,且未观察到细胞毒性作用。它们还有效抑制了α-MSH处理的B16细胞中细胞活性氧(ROS)的产生,提高了超氧化物歧化酶(SOD)活性和谷胱甘肽(GSH)水平。其主要成分的这些活性可达PG-2的80%以上。ROS清除剂N-乙酰-L-半胱氨酸(NAC)对黑色素生成具有类似的抑制作用。

结论

这些结果表明,人参乙酸乙酯提取物对抑制黑色素生成的效果最佳,其主要成分是多酚类化合物,可能通过抑制氧化应激来抑制黑色素生成。本研究为人参皮肤美白作用的活性成分和分子机制提供了新的见解。

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