Objective To study the effect of H2O2 at different concentration on cell proliferation,melanogenesis and tyrosinase (TYR) expression in a co-culture system model of melanocyte (MC) and keratinocyte (KC). Methods A co-culture system model was constructed using the primary human melanocytes and keratinocytes. MTT assay was used to determine cell proliferation, NaOH was used to determine the content of melanin, and real time PCR was used to determine TYR expression. Results A stable coculture system was constructed when melanocyte and keratinocyte were mixed in a proportion of 3 : 1. Cell apoptosis was induced under the treatment of H2O2 at high concentration, and cell proliferation and TYR expression were suppressed (P<0.05). No obvious apoptosis was found when H2O2 was at low concentration (<0.1 mmol/L, P>0.05), and cell proliferation were slightly inhibited, but was improved. TYR expression level showed double trends with different H2O2 level. Conclusions The effect of H2O2 on the co-culture system is very complex with a double trends on melanogenesis.%目的 探讨不同浓度H2O2对黑素细胞(melanocyte,MC)和角质形成细胞(keratinocyte,KC)共培养体系的细胞增殖、黑素合成及酪氨酸酶(tyrosinase,TYR)表达的影响.方法 培养原代人表皮MC和KC,建立共培养模型.采用MTT法测定细胞增殖,NaOH法测定细胞黑素含量,实时PCR方法观察TYR基因的表达.结果 KC和MC以3:1浓度混合时,可形成稳定共培养模型.高浓度H2O2作用下,共培养细胞增殖受到明显抑制(P＜0.05),TYR基因表达显著降低(P＜0.05)；低浓度H2O2(＜0.1 mrmol/L)作用下,细胞增殖受到轻度抑制,TYR基因表达增高(P＜0.05).结论 H2O2对MC与KC共培养体系的作用复杂,对黑素合成功能可呈现双向作用.
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