Lee Ye-Jin, Xu Yang, Hyun Chang-Gu
Jeju Inside Agency and Cosmetic Science Center, Department of Chemistry and Cosmetics, Jeju National University, Jeju 63243, Republic of Korea.
Pharmaceuticals (Basel). 2025 Mar 25;18(4):463. doi: 10.3390/ph18040463.
This study aims to investigate the effects of 5,7-dihydroxy-4-methylcoumarin (5,7D-4MC) on melanogenesis in B16F10 murine melanoma cells and to evaluate its safety as a potential ingredient for functional cosmetics and therapeutic agents targeting pigmentation-related disorders. The cytotoxicity of 5,7D-4MC was assessed using an MTT assay, and melanin content and tyrosinase activity were measured at different concentrations (25, 50, 100 µM). Western blot analyses were conducted to evaluate the expression of key melanogenesis-related proteins (TYR, TRP-1, TRP-2, and MITF) and to investigate the regulation of major signaling pathways, including PKA/cAMP, GSK3β, and PI3K/AKT. Additionally, a human primary skin irritation test was performed on 32 participants to assess the dermatological safety of 5,7D-4MC. 5,7D-4MC did not affect cell viability at concentrations below 100 µM and significantly promoted melanin production in a dose-dependent manner. Tyrosinase activity and the expression levels of melanogenic proteins increased significantly following 5,7D-4MC treatment. PKA and GSK3β pathways were activated, while the PI3K/AKT pathway was downregulated. The skin irritation test showed that 5,7D-4MC exhibited low irritation potential at concentrations of 50 µM and 100 µM. 5,7D-4MC enhances melanogenesis and demonstrates low skin irritation, making it a promising candidate for therapeutic applications in treating hypopigmentation disorders, such as vitiligo, as well as a functional cosmetic ingredient. However, further studies involving human melanocytes and clinical trials are required to validate their efficacy.
本研究旨在探讨5,7 - 二羟基 - 4 - 甲基香豆素（5,7D - 4MC）对B16F10小鼠黑色素瘤细胞黑色素生成的影响，并评估其作为功能性化妆品和针对色素沉着相关疾病的治疗药物潜在成分的安全性。使用MTT法评估5,7D - 4MC的细胞毒性，并在不同浓度（25、50、100 μM）下测量黑色素含量和酪氨酸酶活性。进行蛋白质免疫印迹分析以评估关键黑色素生成相关蛋白（TYR、TRP - 1、TRP - 2和MITF）的表达，并研究主要信号通路的调节，包括PKA/cAMP、GSK3β和PI3K/AKT。此外，对32名参与者进行了人体原发性皮肤刺激试验，以评估5,7D - 4MC的皮肤安全性。5,7D - 4MC在浓度低于100 μM时不影响细胞活力，并以剂量依赖性方式显著促进黑色素生成。5,7D - 4MC处理后，酪氨酸酶活性和黑色素生成蛋白的表达水平显著增加。PKA和GSK3β通路被激活，而PI3K/AKT通路被下调。皮肤刺激试验表明，5,7D - 4MC在50 μM和100 μM浓度下表现出低刺激潜力。5,7D - 4MC增强黑色素生成并表现出低皮肤刺激性，使其成为治疗色素减退性疾病（如白癜风）的治疗应用以及功能性化妆品成分的有前景的候选物。然而，需要进一步涉及人类黑素细胞的研究和临床试验来验证其疗效。
