Huzhou Jiaheng Industrial Co., Ltd, Huzhou, China.
Lanzhou University, Lanzhou, China.
J Cosmet Dermatol. 2024 Sep;23(9):3030-3037. doi: 10.1111/jocd.16370. Epub 2024 Jun 12.
In vitro single-cell experiments may yield inconsistent results compared to clinical trials. To enhance the reliability of cosmetic active ingredient screening, a coculture model of B16F10-HaCaT cells was established in vitro based on the structural characteristics of human skin, thereby improving the credibility of experimental outcomes. Currently, most cosmetic whitening additives primarily target simple efficacy goals such as inhibiting tyrosinase activity or melanin transfer. Therefore, investigating novel and efficient whitening additives has become a prominent research focus.
The aim is to establish an in vitro cell coculture model for more reliable experimental results and investigate the mechanism by which Paeonia lactiflora Pall seeds oil inhibits melanin production and transfer.
The impact of different concentrations of Paeonia lactiflora Pall seeds oil on cocultured cell proliferation rate was assessed using cck8 assay. Tyrosinase inhibition ability in cocultured cells was tested using levodopa as a substrate. Melanin production inhibition ability in coculture cells was evaluated by lysing cells with sodium hydroxide. The effect of Paeonia lactiflora Pall seeds oil on dendrite-related gene expression levels was examined through qPCR analysis. Additionally, Western blotting was employed to study the effect of Paeonia lactiflora Pall seeds oil on dendrite-related protein expression levels.
Different concentrations of Paeonia lactiflora Pall seeds oil did not affect the proliferation activity of cocultured cells. A specific concentration of α-MSH increased cell tyrosinase activity, cellular melanin content, as well as Rac1, Cdc42, and PAR-2 gene and protein expression related to dendritic formation. Treatment with a certain concentration of Paeonia lactiflora Pall seeds oil resulted in decreased tyrosinase activity and melanin content in cells along with downregulated expression levels of Rac1, Cdc42, and PAR-2 genes and proteins associated with dendritic formation.
Paeonia lactiflora Pall seeds oil at specific concentrations exhibits the ability to inhibit tyrosinase activity, decrease melanin content, and possesses the potential to impede melanin transfer.
与临床试验相比，体外单细胞实验的结果可能不一致。为了提高化妆品功效成分筛选的可靠性，本研究基于人皮肤的结构特点，建立了 B16F10-HaCaT 细胞共培养模型，提高了实验结果的可信度。目前，大多数化妆品增白添加剂主要针对抑制酪氨酸酶活性或黑色素转移等简单的功效目标。因此，研究新型、高效的增白添加剂已成为突出的研究重点。
建立体外细胞共培养模型，获得更可靠的实验结果，并研究丹皮酚抑制黑色素生成和转移的作用机制。
采用 CCK8 法检测不同浓度丹皮酚对共培养细胞增殖率的影响，以左旋多巴为底物检测共培养细胞酪氨酸酶抑制能力，用氢氧化钠裂解细胞检测共培养细胞黑色素生成抑制能力，通过 qPCR 分析检测丹皮酚对树突状相关基因表达水平的影响，并用 Western blot 检测丹皮酚对树突状相关蛋白表达水平的影响。
不同浓度丹皮酚对共培养细胞的增殖活性没有影响。特定浓度的 α-MSH 增加了细胞酪氨酸酶活性、细胞黑色素含量，以及 Rac1、Cdc42 和 PAR-2 与树突形成相关的基因和蛋白表达。一定浓度的丹皮酚处理降低了细胞的酪氨酸酶活性和黑色素含量，下调了 Rac1、Cdc42 和 PAR-2 基因和蛋白的表达，这些基因和蛋白与树突形成有关。
特定浓度的丹皮酚具有抑制酪氨酸酶活性、降低黑色素含量和阻止黑色素转移的潜力。
