Jeju Inside Agency and Cosmetic Science Center, Department of Chemistry and Cosmetics, Jeju National University, Jeju 63243, Republic of Korea.
Int J Mol Sci. 2023 Jun 20;24(12):10393. doi: 10.3390/ijms241210393.
In this study, we demonstrated that 2'-hydroxy-3,6'-dimethoxychalcone (3,6'-DMC) alleviated α-MSH-induced melanogenesis and lipopolysaccharides (LPS)-induced inflammation in mouse B16F10 and RAW 264.7 cells. In vitro analysis results showed that the melanin content and intracellular tyrosinase activity were significantly decreased by 3,6'-DMC, without cytotoxicity, via decreases in tyrosinase and the tyrosinase-related protein 1 (TRP-1) and TRP-2 melanogenic proteins, as well as the downregulation of microphthalmia-associated transcription factor (MITF) expression through the upregulation of the phosphorylation of extracellular-signal-regulated kinase (ERK), phosphoinositide 3-kinase (PI3K)/Akt, and glycogen synthase kinase-3β (GSK-3β)/catenin, and downregulation of the phosphorylation of p38, c-Jun N-terminal kinase (JNK), and protein kinase A (PKA). Furthermore, we investigated the effect of 3,6'-DMC on macrophage RAW264.7 cells with LPS stimulation. 3,6'-DMC significantly inhibited LPS-stimulated nitric oxide production. 3,6'-DMC also suppressed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 on the protein level. In addition, 3,6'-DMC decreased the production of the tumor necrosis factor-α and interleukin-6. Successively, our mechanistic studies revealed that 3,6'-DMC also suppressed the LPS-induced phosphorylation of the inhibitor of IκBα, p38MAPK, ERK, and JNK. The Western blot assay results showed that 3,6'-DMC suppresses LPS-induced p65 translocation from cytosol to the nucleus. Finally, the topical applicability of 3,6'-DMC was tested through primary skin irritation, and it was found that 3,6'-DMC, at 5 and 10 μM concentrations, did not cause any adverse effects. Therefore, 3,6'-DMC may provide a potential candidate for preventing and treating melanogenic and inflammatory skin diseases.
在这项研究中，我们证明了 2'-羟基-3,6'-二甲氧基查尔酮（3,6'-DMC）可减轻α-MSH 诱导的黑色素生成和脂多糖（LPS）诱导的小鼠 B16F10 和 RAW 264.7 细胞炎症。体外分析结果表明，3,6'-DMC 通过降低酪氨酸酶和酪氨酸酶相关蛋白 1（TRP-1）和 TRP-2 黑色素生成蛋白，以及下调小眼畸形相关转录因子（MITF）表达，同时上调细胞外信号调节激酶（ERK）、磷酸肌醇 3-激酶（PI3K）/蛋白激酶 B（Akt）和糖原合成酶激酶-3β（GSK-3β）/连环蛋白的磷酸化，显著降低黑素含量和细胞内酪氨酸酶活性，且无细胞毒性。此外，我们研究了 3,6'-DMC 对 LPS 刺激的 RAW264.7 巨噬细胞的影响。3,6'-DMC 显著抑制 LPS 刺激的一氧化氮产生。3,6'-DMC 还抑制了诱导型一氧化氮合酶（iNOS）和环氧化酶（COX）-2 的蛋白表达。此外，3,6'-DMC 降低了肿瘤坏死因子-α和白细胞介素-6 的产生。随后，我们的机制研究表明，3,6'-DMC 还抑制了 LPS 诱导的 IκBα 抑制剂、p38MAPK、ERK 和 JNK 的磷酸化。Western blot 分析结果表明，3,6'-DMC 抑制了 LPS 诱导的 p65 从细胞质向核内的转位。最后，通过原发性皮肤刺激试验测试了 3,6'-DMC 的局部适用性，结果发现 3,6'-DMC 在 5 和 10 μM 浓度下不会引起任何不良反应。因此，3,6'-DMC 可能为预防和治疗黑色素生成和炎症性皮肤病提供了一个潜在的候选药物。
