PDF(Pubmed)
Abstract：
Abnormal melanogenesis can lead to hyperpigmentation. Tyrosinase (TYR), a key rate-limiting enzyme in melanin production, is an important therapeutic target for these disorders. We investigated the TYR inhibitory activity of hydrolysates extracted from the muscle tissue of Takifugu flavidus (TFMH). We used computer-aided virtual screening to identify a novel peptide that potently inhibited melanin synthesis, simulated its binding mode to TYR, and evaluated functional efficacy in vitro and in vivo. TFMH inhibited the diphenolase activities of mTYR, reducing TYR substrate binding activity and effectively inhibiting melanin synthesis. TFMH indirectly reduced cAMP response element-binding protein phosphorylation in vitro by downregulating melanocortin 1 receptor expression, thereby inhibiting expression of the microphthalmia-associated transcription factor, further decreasing TYR, tyrosinase related protein 1, and dopachrome tautomerase expression and ultimately impeding melanin synthesis. In zebrafish, TFMH significantly reduced black spot formation. TFMH (200 μg/mL) decreased zebrafish TYR activity by 43% and melanin content by 52%. Molecular dynamics simulations over 100 ns revealed that the FGFRSP (T-6) peptide stably binds mushroom TYR via hydrogen bonds and ionic interactions. T-6 (400 μmol/L) reduced melanin content in B16F10 melanoma cells by 71% and TYR activity by 79%. In zebrafish, T-6 (200 μmol/L) inhibited melanin production by 64%. TFMH and T-6 exhibit good potential for the development of natural skin-whitening cosmetic products.
摘要：
黑素生成异常可导致色素沉着过度。酪氨酸酶(TYR),黑色素生产中的关键限速酶，是这些疾病的重要治疗靶点。我们研究了从Takifuguflavidus（TFMH）的肌肉组织中提取的水解产物的TYR抑制活性。我们使用计算机辅助虚拟筛选来鉴定一种有效抑制黑色素合成的新型肽，模拟了它与TYR的绑定模式，并在体外和体内评估功能功效。TFMH抑制mTYR的二酚酶活性，降低TYR底物结合活性，有效抑制黑色素合成。TFMH通过下调黑皮质素1受体表达，在体外间接降低cAMP反应元件结合蛋白磷酸化，从而抑制小眼症相关转录因子的表达，进一步降低TYR，酪氨酸酶相关蛋白1和多巴色素互变异构体的表达并最终阻碍黑色素的合成。在斑马鱼中,TFMH显著减少黑点形成。TFMH（200μg/mL）使斑马鱼TYR活性降低43％，黑色素含量降低52％。超过100ns的分子动力学模拟表明，FGFRSP（T-6）肽通过氢键和离子相互作用稳定地结合蘑菇TYR。T-6（400μmol/L）使B16F10黑素瘤细胞中的黑色素含量降低71％，TYR活性降低79％。在斑马鱼中,T-6(200μmol/L)抑制黑色素生成64%。TFMH和T-6显示出开发天然皮肤美白化妆品的良好潜力。
