不同纯度高良姜素对A375-HaCaT共培养模型中黑素细胞增殖及黑素合成的影响

Effect of Different Purity Galangin on Melanocyte Proliferation and Melanin Synthesis of A375-HaCaT Co-culture System

目的研究不同纯度高良姜素对人黑色素瘤细胞-人永生化角质形成细胞(A375-HaCaT)共培养模型中黑素细胞增殖及黑素合成的影响。方法利用Transwell技术建立A375-HaCaT共培养体系,以0.1、0.5、1.0、5.0、10μg/mL纯度为70%和90%的高良姜素作用于共培养模型。采用MTT比色法检测细胞增殖,NaOH裂解法测定黑素含量,多巴氧化法测定酪氨酸酶活性,ELISA法检测HaCaT细胞因子干细胞生长因子(SCF)、内皮素(ET)-1的表达。结果 A375-HaCaT共培养模型中的A375细胞生长良好,0.1、0.5、1.0、5.0、10μg/mL纯度为70%和90%的高良姜素对共培养体系中的A375细胞增殖、酪氨酸酶活性及黑素合成均具有上调作用。各纯度高良姜素浓度≥0.5μg/mL时,能够提高HaCaT细胞中的ET-1和SCF等细胞因子的表达水平,且纯度70%的高良姜素作用强度优于90%的高良姜素。结论高良姜素能显著促进A375-HaCaT共培养模型中A375的细胞增殖及黑素合成,其作用机制可能与高良姜素上调HaCaT细胞中的ET-1和SCF等细胞因子的表达水平有关。

Objective To investigate the effect of Galangin of different purity on melanocyte proliferation and melanin synthesis of A375-HaCaT co-culture system. Methods The A375-HaCaT co-culture system was established with Transwell technology, and 0.1, 0.5, 1.0, 5.0, 10 μg/mL Galangin of the purity of 70%and 90%was used to act on the co-culture system. Cell proliferation, melanin content and tyrosinase of A375 were measured by MTT assay, NaOH lysis assay and L-DOPA oxidation assay respectively. The cytokines such as ET-1 and SCF in HaCaT were detected by ELISA. Results A375 cells in co-culture system grew well, and 0.1, 0.5, 1.0, 5.0, 10 μg/mL Galangin of the purity of 70% and 90% had up-regulation effect on cell proliferation, melanin content and tyrosinase of A375. Moreover, Galangin could increase the expression level of ET-1 and SCF of HaCaT at more than 0.5 μg/mL, and the effect of Galangin of 70% purity was better than 90% purity. Conclusion Galangin could promote the cell proliferation, melanin content and tyrosinase of A375, and mechanism of the pathways is probably related to the up-regulation on the expression of ET-1 and SCF of HaCaT.