白芨水提物对黑素瘤与角质形成细胞共培养模型黑素合成的影响

Influence of Bletilla Water Extract of Melanoma Cells and Keratinocytes on Melanin Synthesis in Co-culture Model

目的:利用人表皮黑素瘤细胞(A375细胞)与人永生化角质形成细胞(HaCat细胞)共培养模型,观察白芨水提物对该模型中黑素合成的影响。方法:构建A375细胞与HaCat细胞共培养模型,采用氢氧化钠裂解法、MTT法和多巴氧化法分别检测白芨水提物对黑素含量、细胞增殖和酪氨酸酶(TYR)活性的影响。结果:与阴性对照组比较,2 g/L、1 g/L、0.5 g/L、0.25 g/L白芨水提物对黑素合成有显著抑制作用(P<0.01),2 g/L、1 g/L组对黑素合成的抑制作用与阳性对照药物熊果苷组相当;白芨水提物对TYR活性有显著抑制作用(P<0.01),其中0.5 g/L、0.25 g/L和0.125 g/L组与阳性对照药物熊果苷组的作用相当。结论:白芨水提取物可以通过抑制TYR活性抑制A375细胞与Hacat细胞共培养体系的黑素合成,为临床使用白芨治疗黄褐斑提供了实验依据。

Objective：Using human epidermal melanoma cells （A375 cells） and immortalized human keratinocytes （HaCat cells） in co-culture model to observe the effect of the aqueous extract of Bletilla on melanin synthesis.Methods：Construction of A375 cells and HaCat cell co-culture model with sodium hydroxide decomposition method,influence of Bletilla water extract melanoma cells and keratinocytes on melanin synthesis,cell proliferation and tyrosinase （TYR） activity were determined by MTT method and dopamine oxidation.Results：Compared with negative control group,2 g/L,1 g/ L,0.5 g/L,0.25 g/L bletilla aqueous extract had a significant inhibition of melanin synthesis （P 〈 0.01）,the inhibition effects of group 2 g/L,1 g/L were equal to those of positive control drug arbutin group; Bletilla aqueous extract had significant inhibitory effect on activity of TYR （P 〈 0.01）,of which 0.5 g/L,0.25 g and 0.125 g/L/L group were equal to those of positive control drug arbutin group.Conclusions：Bletilla water extract can inhibit TYR activity of A375 cells and melanin synthesis in HaCat cell co-culture system and provide an experimental reference for clinical use of bletilla treatment of chloasma.