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目的:考察山茱萸水提取物对体外构建的人黑素瘤细胞(A375)与角质形成细胞(Hacat)单层混合共培养模型黑素合成的影响。方法:将山茱萸水提取物制备成含生药0.75～12 g.L-15个质量浓度,细胞给药,通过MTT法、NaOH裂解法、多巴氧化法分别测定给药前后A375细胞与Hacat细胞共培养模型的细胞活力、黑素含量和酪氨酸酶活性。结果:与空白对照组相比,6,3,1.5,0.75 g.L-1山茱萸水提取物对共培养体系的细胞增殖率影响不大,对共培养体系黑素生成均有显著差异(P<0.01),对共培养体系酪氨酸酶活性有差异(P<0.05或P<0.01);与阳性对照药物熊果苷相比,6,3 g.L-1山茱萸水提取物对共培养体系黑素生成有显著差异(P<0.01),作用强于熊果苷,1.5,0.75 g.L-1山茱萸水提取物对共培养体系黑素生成无差异;与阳性对照药物熊果苷相比,6,3,1.5,0.75 g.L-1山茱萸水提取物对共培养体系酪氨酸酶活性无差异。结论:山茱萸水提取物能够通过抑制酪氨酸酶活性来抑制A375细胞和Hacat细胞共培养体系的黑素合成,为临床使用山茱萸治疗黄褐斑提供了实验依据。
Objective: To investigate the effect of Fructus corni aqueous extract on the melanogenesis of a mixed monolayer culture model of human melanoma cells (A375) and keratinocytes (Hacat) in vitro. Methods: The Fructus Corni water extract was prepared with 0.75 ~ 12 gL-15concentration of crude drug, and the cells were administrated. The co-culture model of A375 cells and Hacat cells were determined by MTT method, NaOH cleavage method and dopa oxidation method Cell viability, melanin content and tyrosinase activity. Results: Compared with the blank control group, the water extract of 6, 3, 1.5, and 0.75 g L-1 of Fructus Corni did not significantly affect the cell proliferation rate of the co-culture system, but had significant difference on the melanogenesis of the co-culture system ), The tyrosinase activity of the co-culture system was different (P <0.05 or P <0.01); compared with the positive control drug arbutin, 6,3 gL-1 of Fructus corni water extract of co-culture system melanogenesis (P <0.01), stronger than arbutin, 1.5,0.75 gL-1 of dogwood water extract no difference in melanogenesis of co-culture system; compared with the positive control drug arbutin, 6,3, 1.5,0.75 gL-1 dogwood water extract on the co-cultivation system tyrosinase activity no difference. Conclusion: Fructus corni aqueous extract can inhibit the melanogenesis of A375 cells and Hacat cell co-culture system by inhibiting the tyrosinase activity, which provides experimental evidence for the clinical use of dogwood for the treatment of melasma.