Abstract：
BACKGROUND: The pursuit for safe and efficacious skin-whitening agents has prompted a dedicated exploration of plant-derived compounds. Notably, Tagetes erecta L. flowers have been used as a medicinal extract and possessed in vitro mushroom tyrosinase activity. However, whether polyphenol-enriched fraction extracted from T. erecta L. flowers (TE) regulates melanogenesis within cellular and animal models has not yet been investigated.
OBJECTIVE: This study aimed to investigate the effect of TE as a prospective inhibitor of melanogenesis.
METHODS: Through advanced UPLC-QTof/MS analysis, the components of TE were analyzed. Anti-melanogenic effects of TE were evaluated in α-melanocyte-stimulating hormone (α-MSH)-stimulated B16F10 melanoma cells by measuring cell viability assay, extracellular and intracellular melanin biosynthesis, cyclic adenosine monophosphate (cAMP) production, and melanogenesis-related gene and protein expression. Zebrafish larvae were employed for in vivo studies, assessing both heart rate and melanogenesis. Furthermore, molecular docking analyses were employed to predict the interaction between TE components and the melanocortin 1 receptor (MC1R). Direct binding activity of TE components to MC1R was compared with [Nle4, d-Phe7]-MSH (NDP-MSH).
RESULTS: TE was found to contain significant phenolic compounds such as patulitrin, quercetagetin, kaempferol, patuletin, and isorhamnetin. This study revealed that TE effectively inhibits melanin biosynthesis in both in vitro and in vivo models. This inhibition was attributed to interference of TE with the cAMP-cAMP response element-binding protein (CREB)-microphthalmia-associated transcription factor (MITF)-tyrosinase pathway, which plays a pivotal role in regulating melanogenesis. Importantly, TE exhibited the remarkable ability to curtail α-MSH-induced melanogenesis in zebrafish larvae without impacting heart rates. Molecular docking analyses predicted that the components of TE possibly interact with the melanocortin 1 receptor, suggesting their role as potential inhibitors of melanin biosynthesis. However, through the direct binding activity compared with NDP-MSH, any TE components did not directly bind to MC1R, suggesting that TE inhibits α-MSH-induced melanogenesis by inhibiting the cAMP-mediated intracellular signaling pathway. The assessment of anti-melanogenic activity, conducted both in vitro and in vivo, revealed that patulitrin and patuletin exhibited significant inhibitory effects on melanin formation, highlighting their potency as major contributors.
CONCLUSIONS: This investigation demonstrated the considerable potential of TE as a natural remedy endowed with remarkable anti-melanogenic properties. The demonstrated capacity of TE to attenuate melanin production by modulating the cAMP-CREB-MITF-tyrosinase pathway underscores its central role in management of disorders associated with excessive pigmentation. Importantly, the implications of these findings extend to the cosmetics industry, where TE emerges as a prospective and valuable ingredient for the formulation of skin-whitening products. The elucidated interactions between TE components and MC1R not only provide insight into a potential mechanism of action but also elevate the significance of this study. In summary, this study not only contributes to our comprehension of pigmentation-related conditions but also firmly establishes TE as a secure and natural strategy for the regulation of melanin production. The innovative aspects of TE propel it into the forefront of potential interventions, marking a noteworthy advancement in the pursuit of effective and safe solutions for pigmentation disorders.
OBJECTIVE: This study aimed to investigate the effect of TE as a prospective inhibitor of melanogenesis.
METHODS: Through advanced UPLC-QTof/MS analysis, the components of TE were analyzed. Anti-melanogenic effects of TE were evaluated in α-melanocyte-stimulating hormone (α-MSH)-stimulated B16F10 melanoma cells by measuring cell viability assay, extracellular and intracellular melanin biosynthesis, cyclic adenosine monophosphate (cAMP) production, and melanogenesis-related gene and protein expression. Zebrafish larvae were employed for in vivo studies, assessing both heart rate and melanogenesis. Furthermore, molecular docking analyses were employed to predict the interaction between TE components and the melanocortin 1 receptor (MC1R). Direct binding activity of TE components to MC1R was compared with [Nle4, d-Phe7]-MSH (NDP-MSH).
RESULTS: TE was found to contain significant phenolic compounds such as patulitrin, quercetagetin, kaempferol, patuletin, and isorhamnetin. This study revealed that TE effectively inhibits melanin biosynthesis in both in vitro and in vivo models. This inhibition was attributed to interference of TE with the cAMP-cAMP response element-binding protein (CREB)-microphthalmia-associated transcription factor (MITF)-tyrosinase pathway, which plays a pivotal role in regulating melanogenesis. Importantly, TE exhibited the remarkable ability to curtail α-MSH-induced melanogenesis in zebrafish larvae without impacting heart rates. Molecular docking analyses predicted that the components of TE possibly interact with the melanocortin 1 receptor, suggesting their role as potential inhibitors of melanin biosynthesis. However, through the direct binding activity compared with NDP-MSH, any TE components did not directly bind to MC1R, suggesting that TE inhibits α-MSH-induced melanogenesis by inhibiting the cAMP-mediated intracellular signaling pathway. The assessment of anti-melanogenic activity, conducted both in vitro and in vivo, revealed that patulitrin and patuletin exhibited significant inhibitory effects on melanin formation, highlighting their potency as major contributors.
CONCLUSIONS: This investigation demonstrated the considerable potential of TE as a natural remedy endowed with remarkable anti-melanogenic properties. The demonstrated capacity of TE to attenuate melanin production by modulating the cAMP-CREB-MITF-tyrosinase pathway underscores its central role in management of disorders associated with excessive pigmentation. Importantly, the implications of these findings extend to the cosmetics industry, where TE emerges as a prospective and valuable ingredient for the formulation of skin-whitening products. The elucidated interactions between TE components and MC1R not only provide insight into a potential mechanism of action but also elevate the significance of this study. In summary, this study not only contributes to our comprehension of pigmentation-related conditions but also firmly establishes TE as a secure and natural strategy for the regulation of melanin production. The innovative aspects of TE propel it into the forefront of potential interventions, marking a noteworthy advancement in the pursuit of effective and safe solutions for pigmentation disorders.
摘要：
背景：对安全有效的皮肤增白剂的追求促使了对植物衍生化合物的专门探索。值得注意的是,万寿菊花已被用作药用提取物，并具有体外蘑菇酪氨酸酶活性。然而,是否从T.eriptaL.花（TE）中提取的富含多酚的部分调节细胞和动物模型中的黑素生成尚未研究。
目的：本研究旨在研究TE作为黑素生成的前瞻性抑制剂的作用。
方法：通过先进的UPLC-QTof/MS分析，对TE的成分进行了分析。在α-黑素细胞刺激激素（α-MSH）刺激的B16F10黑色素瘤细胞中，通过测量细胞活力测定法评估了TE的抗黑色素生成作用。细胞外和细胞内黑色素生物合成，环磷酸腺苷（cAMP）的生产，和黑色素生成相关基因和蛋白质表达。斑马鱼幼虫用于体内研究，评估心率和黑色素生成。此外，分子对接分析用于预测TE组分与黑皮质素1受体（MC1R）之间的相互作用。将TE组分与MC1R的直接结合活性与[Nle4,d-Phe7]-MSH(NDP-MSH)进行比较。
结果：TE被发现含有显著的酚类化合物，如patulitrin，槲皮素,山奈酚,patutinetin,和isorhametin.这项研究表明，TE在体外和体内模型中都能有效抑制黑色素的生物合成。这种抑制作用归因于TE对cAMP-cAMP反应元件结合蛋白（CREB）-小眼症相关转录因子（MITF）-酪氨酸酶途径的干扰，在调节黑色素生成中起着关键作用。重要的是,TE在斑马鱼幼虫中表现出明显的减少α-MSH诱导的黑素生成的能力，而不会影响心率。分子对接分析表明，TE的成分可能与黑皮质素1受体相互作用，表明它们作为黑色素生物合成的潜在抑制剂的作用。然而,通过与NDP-MSH相比的直接结合活性，任何TE组件都不直接与MC1R结合，提示TE通过抑制cAMP介导的细胞内信号通路抑制α-MSH诱导的黑素生成。抗黑色素生成活性的评估，在体外和体内进行，显示patulitrin和patuletin对黑色素形成具有显著的抑制作用，强调他们作为主要贡献者的能力。
结论：这项研究证明了TE作为一种天然药物具有显著的抗黑色素生成特性的巨大潜力。证实的TE通过调节cAMP-CREB-MITF-酪氨酸酶途径来减弱黑色素产生的能力强调了其在与过度色素沉着相关的疾病的管理中的核心作用。重要的是,这些发现的影响延伸到化妆品行业，其中TE成为皮肤美白产品配方的潜在和有价值的成分。阐明的TE成分与MC1R之间的相互作用不仅提供了对潜在作用机制的了解，而且提高了本研究的意义。总之,这项研究不仅有助于我们对色素沉着相关疾病的理解,而且还牢固地确立了TE作为调节黑色素产生的安全和自然策略.TE的创新方面推动其进入潜在干预措施的最前沿，标志着在寻求有效和安全的色素沉着障碍解决方案方面取得了显著进展。
目的：本研究旨在研究TE作为黑素生成的前瞻性抑制剂的作用。
方法：通过先进的UPLC-QTof/MS分析，对TE的成分进行了分析。在α-黑素细胞刺激激素（α-MSH）刺激的B16F10黑色素瘤细胞中，通过测量细胞活力测定法评估了TE的抗黑色素生成作用。细胞外和细胞内黑色素生物合成，环磷酸腺苷（cAMP）的生产，和黑色素生成相关基因和蛋白质表达。斑马鱼幼虫用于体内研究，评估心率和黑色素生成。此外，分子对接分析用于预测TE组分与黑皮质素1受体（MC1R）之间的相互作用。将TE组分与MC1R的直接结合活性与[Nle4,d-Phe7]-MSH(NDP-MSH)进行比较。
结果：TE被发现含有显著的酚类化合物，如patulitrin，槲皮素,山奈酚,patutinetin,和isorhametin.这项研究表明，TE在体外和体内模型中都能有效抑制黑色素的生物合成。这种抑制作用归因于TE对cAMP-cAMP反应元件结合蛋白（CREB）-小眼症相关转录因子（MITF）-酪氨酸酶途径的干扰，在调节黑色素生成中起着关键作用。重要的是,TE在斑马鱼幼虫中表现出明显的减少α-MSH诱导的黑素生成的能力，而不会影响心率。分子对接分析表明，TE的成分可能与黑皮质素1受体相互作用，表明它们作为黑色素生物合成的潜在抑制剂的作用。然而,通过与NDP-MSH相比的直接结合活性，任何TE组件都不直接与MC1R结合，提示TE通过抑制cAMP介导的细胞内信号通路抑制α-MSH诱导的黑素生成。抗黑色素生成活性的评估，在体外和体内进行，显示patulitrin和patuletin对黑色素形成具有显著的抑制作用，强调他们作为主要贡献者的能力。
结论：这项研究证明了TE作为一种天然药物具有显著的抗黑色素生成特性的巨大潜力。证实的TE通过调节cAMP-CREB-MITF-酪氨酸酶途径来减弱黑色素产生的能力强调了其在与过度色素沉着相关的疾病的管理中的核心作用。重要的是,这些发现的影响延伸到化妆品行业，其中TE成为皮肤美白产品配方的潜在和有价值的成分。阐明的TE成分与MC1R之间的相互作用不仅提供了对潜在作用机制的了解，而且提高了本研究的意义。总之,这项研究不仅有助于我们对色素沉着相关疾病的理解,而且还牢固地确立了TE作为调节黑色素产生的安全和自然策略.TE的创新方面推动其进入潜在干预措施的最前沿，标志着在寻求有效和安全的色素沉着障碍解决方案方面取得了显著进展。
