Wang Hui Miao, Qu Li Qun, Ng Jerome P L, Zeng Wu, Yu Lu, Song Lin Lin, Wong Vincent Kam Wai, Xia Cheng Lai, Law Betty Yuen Kwan
Dr. Neher's Biophysics Laboratory for Innovative Drug Discovery, State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China.
Affiliated Foshan Maternity & Child Healthcare Hospital, Southern Medical University, Foshan, Guangdong 528000, China; School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510150, China.
Phytomedicine. 2022 Apr;98:153941. doi: 10.1016/j.phymed.2022.153941. Epub 2022 Jan 19.
5-demethylnobiletin is a natural polymethoxyflavone which is isolated from the extract of citrus fruits peels. It exhibits a broad spectrum of biological activities such as anti-cancer, anti-inflammatory, cardiovascular protective and neuroprotective effects, however, its effect in melanogenesis remains uninvestigated.
Melanin synthesis is a very important biological process in curing disease such as vitiligo with depigmentation on the skin. In the current work, we aim to confirm the bioactivity and mechanism of 5-demethylnobiletin in stimulating melanogenesis.
To confirm the mechanistic role of 5-demethylnobiletin in enhancing melanogenesis, its effect on the activity of tyrosinase, together with the level of microphthalmia-associated transcription factor (MITF), Trp-1, Trp-2, melanocyte-specific marker protein PMEL17, Rab27a, Melanophilin and Myosin VA were studied in B16F10 melanoma cells.
Multiple biological assays on melanogenesis-associated proteins such as melanin content detection, tyrosinase activity colorimetric assay, qPCR, western blot analysis, dual-luciferase reporter assay, cAMP activity assay and Fontana-Masson ammoniacal silver staining were used to confirm the role of 5-demethylnobiletin in stimulating melanin synthesis and the transportation of melanosomes.
As confirmed by multiple biological assays, 5-demethylnobiletin is found to stimulate dendrite structure formation in cells, melanin synthesis and the transportation of melanosomes, via inducing the phosphorylation of cAMP response element-binding protein (CREB) and increasing the intracellular levels of cAMP in vitro through the PKA-dependent pathway.
The findings suggested that 5-demethylnobiletin may be considered as a potential natural product candidate for patients with pigment disorder.
5-去甲基川陈皮素是一种从柑橘类水果果皮提取物中分离得到的天然多甲氧基黄酮。它具有广泛的生物活性，如抗癌、抗炎、心血管保护和神经保护作用，然而，其在黑色素生成中的作用仍未得到研究。
黑色素合成是治疗白癜风等皮肤色素脱失疾病的一个非常重要的生物学过程。在当前的工作中，我们旨在确认5-去甲基川陈皮素刺激黑色素生成的生物活性和机制。
为了确认5-去甲基川陈皮素在增强黑色素生成中的作用机制，研究了其对B16F10黑色素瘤细胞中酪氨酸酶活性以及小眼畸形相关转录因子（MITF）、Trp-1、Trp-2、黑色素细胞特异性标记蛋白PMEL17、Rab27a、黑色素亲和素和肌球蛋白VA水平的影响。
采用多种与黑色素生成相关蛋白的生物学检测方法，如黑色素含量检测、酪氨酸酶活性比色法、qPCR、蛋白质免疫印迹分析、双荧光素酶报告基因检测、cAMP活性检测和Fontana-Masson氨银染色，以确认5-去甲基川陈皮素在刺激黑色素合成和黑素小体转运中的作用。
通过多种生物学检测证实，5-去甲基川陈皮素在体外通过PKA依赖途径诱导cAMP反应元件结合蛋白（CREB）磷酸化并增加细胞内cAMP水平，从而刺激细胞中树突结构形成、黑色素合成和黑素小体转运。
研究结果表明，5-去甲基川陈皮素可能被认为是一种治疗色素紊乱患者的潜在天然产物候选物。
