Jeju Inside Agency and Cosmetic Science Center, Department of Chemistry and Cosmetics, Jeju National University, Jeju 63243, Republic of Korea.
Molecules. 2023 Mar 29;28(7):3039. doi: 10.3390/molecules28073039.
The objectives of this study were to investigate the melanogenetic potentials of the naturally occurring 7-hydroxy coumarin derivatives 7-hydroxy 5,6-dimethoxycoumarin (7H-5,6DM), 7-hydroxy 6,8-dimethoxycoumarin (7H-6,8DM), 7-hydroxy 6-methoxycoumarin (7H-6M), and 7-hydroxy 4-methylcoumarin (7H-4M) in the melanogenic cells model for murine B16F10 melanoma cells. The initial results indicated that melanin production and intracellular tyrosinase activity were significantly stimulated by 7H-4M but not by 7H-5,6DM, 7H-6,8DM, or 7H-6M. Therefore, our present study further investigated the melanogenic effects of 7H-4M in B16-F10 cells, as well as its mechanisms of action. In a concentration-dependent manner, 7H-4M increased intracellular tyrosinase activity, leading to the accumulation of melanin without affecting the viability of B16-F10 cells. Our study further investigated the effects of 7H-4M on melanogenesis, including its ability to promote tyrosinase activity, increase melanin content, and activate molecular signaling pathways. The results indicate that 7H-4M effectively stimulated tyrosinase activity and significantly increased the expression of melanin synthesis-associated proteins, such as microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein-1 (TRP1), and TRP2. Based on our findings, we can conclude that 7H-4M has the ability to activate the melanogenesis process through the upregulation of cAMP-dependent protein kinase (PKA) and the cAMP response element-binding protein (CREB). Additionally, our study showed that 7H-4M induced melanogenic effects by downregulating the extracellular signal-regulated kinase (ERK) and the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt)/glycogen synthesis kinase-3β (GSK-3β) cascades, while upregulating the JNK and p38 signaling pathways. Finally, the potential of using 7H-4M in topical applications was tested through primary human skin irritation tests. During these tests, no adverse reactions were induced by 7H-4M. In summary, our results indicate that 7H-4M regulates melanogenesis through various signaling pathways such as GSK3β/β-catenin, AKT, PKA/CREB, and MAPK. These findings suggest that 7H-4M has the potential to prevent the development of pigmentation diseases.
本研究旨在探究天然存在的 7-羟基香豆素衍生物 7-羟基-5,6-二甲氧基香豆素(7H-5,6DM)、7-羟基-6,8-二甲氧基香豆素(7H-6,8DM)、7-羟基-6-甲氧基香豆素(7H-6M)和 7-羟基-4-甲基香豆素(7H-4M)在黑素细胞生成模型中对鼠 B16F10 黑素瘤细胞的黑素生成潜能。初步结果表明,7H-4M 可显著刺激黑色素生成和细胞内酪氨酸酶活性,但 7H-5,6DM、7H-6,8DM 或 7H-6M 则无此作用。因此,本研究进一步探究了 7H-4M 在 B16-F10 细胞中的黑素生成作用及其作用机制。7H-4M 以浓度依赖的方式增加细胞内酪氨酸酶活性,导致黑色素积累,而不影响 B16-F10 细胞的活力。本研究进一步探究了 7H-4M 对黑素生成的影响,包括其促进酪氨酸酶活性、增加黑色素含量和激活分子信号通路的能力。结果表明,7H-4M 可有效刺激酪氨酸酶活性,并显著增加黑色素合成相关蛋白的表达,如小眼畸形相关转录因子(MITF)、酪氨酸酶、酪氨酸酶相关蛋白-1(TRP1)和 TRP2。基于我们的发现,我们可以得出结论,7H-4M 通过上调环磷酸腺苷(cAMP)依赖性蛋白激酶(PKA)和 cAMP 反应元件结合蛋白(CREB)来激活黑素生成过程。此外,我们的研究表明,7H-4M 通过下调细胞外信号调节激酶(ERK)和磷脂酰肌醇 3 激酶(PI3K)/蛋白激酶 B(Akt)/糖原合成激酶-3β(GSK-3β)级联,同时上调 JNK 和 p38 信号通路,诱导黑素生成效应。最后,通过对原代人皮肤刺激试验测试了 7H-4M 在局部应用中的潜力。在这些试验中,7H-4M 未引起不良反应。总之,我们的结果表明,7H-4M 通过 GSK3β/β-连环蛋白、AKT、PKA/CREB 和 MAPK 等多种信号通路调节黑素生成。这些发现表明,7H-4M 具有预防色素沉着疾病发展的潜力。