Department of Cosmetics Engineering, Konkuk University.
Department of Obstetrics and Gynecology, Konkuk University School of Medicine, Seoul, Republic of Korea.
Pharmazie. 2023 Apr 15;78(1):6-12. doi: 10.1691/ph.2023.2490.
The aim of the present study was to investigate the potential inhibitory effects of nodakenin, a coumarin glucoside derivative from the root extract of Nakai (AGN), on melanogenesis and its underlying mechanisms in B16F10 melanoma cells. The inhibitory effects of nodakenin on melanogenesis were evaluated by determining melanin contents and tyrosinase activity in α -melanocyte stimulating hormone (α-MSH)-treated B16F10 melanoma cells. The mechanisms associated with the anti-pigmentation effect of nodakenin were investigated by quantitative real-time PCR and immunoblotting analysis. Using the UVB-irradiated conditioned media culture system and UVB-irradiated co-cultivation system of HaCaT keratinocytes and B16F10 melanoma cells mimicking melanin biosynthesis, the effect of nodakenin on melanin production was evaluated. Melanin content analysis showed that nodakenin decreased cellular melanin biosynthesis in α-MSH-treated B16F10 cells. Immunoblotting revealed that CREB phosphorylation, MITF, a mastering transcription factor of melanogenesis and its downstream genes tyrosinase, tyrosinase-related protein 1, and tyrosinase-related protein 2 were downregulated by nodakenin in a dose-dependent manner. Interestingly, nodakenin did not affect the phosphorylation of PKA and p38 MAPK but the phosphorylation of ERK1/2 and MSK1. In addition, the inhibition of melanin accumulation by nodakenin in the UVB-irradiated conditioned media culture system and UVB-irradiated co-cultivation system of HaCaT and B16F10 cells suggests that nodakenin has potential as an anti-pigmentation activity. These data suggest that nodakenin inhibits the melanogenesis in B16F10 cells by interfering the ERK/ MSK1/CREB axis and thus preventing MITF expression.
本研究旨在探讨从 Nakai（AGN）根提取物中分离得到的香豆素葡萄糖苷衍生物——虎杖苷对黑色素生成的潜在抑制作用及其在 B16F10 黑素瘤细胞中的作用机制。通过测定 α-促黑素细胞激素（α-MSH）处理的 B16F10 黑素瘤细胞中黑色素含量和酪氨酸酶活性来评估虎杖苷对黑色素生成的抑制作用。通过实时定量 PCR 和免疫印迹分析研究了虎杖苷抗色素沉着作用的机制。采用 UVB 照射条件培养基培养系统和 HaCaT 角质形成细胞与 B16F10 黑素瘤细胞共培养的 UVB 照射系统模拟黑色素生物合成，评估了虎杖苷对黑色素生成的影响。黑色素含量分析表明，虎杖苷降低了 α-MSH 处理的 B16F10 细胞中的细胞黑色素生物合成。免疫印迹显示，虎杖苷以剂量依赖性方式下调 CREB 磷酸化、MITF（黑色素生成的主要转录因子）及其下游基因酪氨酸酶、酪氨酸酶相关蛋白 1 和酪氨酸酶相关蛋白 2。有趣的是，虎杖苷不影响 PKA 和 p38 MAPK 的磷酸化，但影响 ERK1/2 和 MSK1 的磷酸化。此外，虎杖苷在 UVB 照射条件培养基培养系统和 UVB 照射 HaCaT 和 B16F10 细胞共培养系统中抑制黑色素积累，表明虎杖苷具有作为抗色素沉着活性的潜力。这些数据表明，虎杖苷通过干扰 ERK/MSK1/CREB 轴抑制 B16F10 细胞中的黑色素生成，从而阻止 MITF 表达。
