熊果苷对A375细胞与HACAT共培养模型中黑素合成的影响

Effect of Arbutin on the in vitro model of the Melanoma Cells and keratinocyte co-culture system

目的:体外构建研究色素问题的黑素瘤细胞(A375)与角质形成细胞(HACAT)直接接触的共培养模型,利用熊果苷对此共培养体系进行验证。方法:MEM培养基分别培养A375细胞与HACAT细胞,A375细胞与HACAT以1:2的比例共培养,构建黑素瘤细胞与角质形成细胞直接接触的共培养模型;将不同浓度熊果苷作用于此模型,用MTT法、多巴氧化法及NaOH裂解法检测熊果苷对细胞增殖、酪氨酸酶活性以及黑素合成的影响。结果:A375细胞与HACAT能共同存活,1.0mg/ml、0.5mg/ml和0.2mg/ml熊果苷对共培养细胞的增殖、黑素细胞酪氨酸酶活性及黑素合成均有较强的剂量相关的抑制作用,与对照组相比差异有统计学意义(P<0.05或P<0.01)。结论:成功构建了A375细胞与HACAT细胞直接接触的共培养模型;脱色药熊果苷作用于本模型后引起酪氨酸酶及黑素的变化与预期结果一致;本模型为进一步研究色素问题奠定了基础。

Objective To Establish the in vitro co-culture model of melanoma cells and keratinocytes,and observe the effect of Arbutin on pigmentation in co-culture model of melanoma cells and keratinocytes. Methods We cultivated the melanoma cells and keratinocytes respectively and then constructed the mixed cultivating model of them.After arbutin was added to the model,celludar proliferation,tyrosinase activity and melanin content were measured by MTT assay,L-DOPA oxidation assay and NaOH assay respectively. Results The in vitro co-culture model of melanoma cells and keratinocytes was established successfully The proliferation of co -culture cells,tyrosinase activity and the melanin synthesis were markedly suppressed by arbutin in a dose-dependent manner.The significant suppression was observed with 1.0mg/ml,0.5mg/ml,0.2mg/ml of arbutin than that with control. Conclusion We have successfully constructed the co-culture model of melanoma cells and keratinocytes,Arbutin known to inhibit melanogenesis were examined using the co-culture model and the results were consistent with expectations. This model provide requirement for further study.