摘要
目的探讨不同浓度紫铆花素对黑素瘤细胞A375与角质形成细胞Hacat共培养体系细胞增殖、酪氨酸酶(TYR)活性、黑色素合成及酪氨酸相关蛋白1(TRP-1)、酪氨酸相关蛋白2(TRP-2) mRNA表达的影响。方法建立黑色素瘤细胞A375与角质形成细胞Hacat共培养体系,以不同浓度紫铆花素处理共培养体系48 h,采用噻唑蓝(MTT)法测定细胞增殖,氢氧化钠(Na OH)裂解法测定细胞黑色素含量,多巴氧化速率法测定黑色素细胞TYR活性,实时荧光聚合酶链反应(Real-time PCR)法检测TRP-1、TRP-2基因表达。结果与细胞共培养体系(即阴性对照组)比较,1.0,5.0μg·mL~(-1)紫铆花素与共培养体系细胞共孵育48 h后,可明显促进共培养细胞增殖(P<0.05或P<0.01),促进黑色素细胞TYR活性(P<0.05或P<0.01);在0.5~50μg·mL~(-1)范围内,紫铆花素可促进共培养体系黑色素合成(P<0.05或P<0.01);在0.1~0.5μg·mL~(-1)范围内,紫铆花素各剂量组TRP-1 mRNA表达升高(P<0.05,P<0.01);在0.5~5.0μg·mL~(-1)范围内,紫铆花素各剂量组TRP-2 mRNA表达升高(P<0.01)。讨论紫铆花素可能通过促进TYR活性和TRP-1、TRP-2mRNA表达增强共培养体系黑色素合成功能。
Objective To investigate the effects of butein on the proliferation,tyrosinase(TYR) activity,melanin synthesis and the mRNA expression of tyrosine-related protein 1(TRP-1) and tyrosine-related protein 2(TRP-2) in the in vitro co-culture model of keratinocytes and melanoma cells.Methods The in vitro co-culture model of A375 and Hacat cells was established.Melanosome synthesis changes induced by different concentration of butein in mixed cultures of A375 and Hacat cells were observed 48 h after treatment,respectively.The cell proliferation rate was detected by MTT,the melanin content was measured with a sodium hydroxide solubilization method,the activity of TYR was estimated by measuring the rate of oxidation of L-dopa,and the mRNA expression of TRP-1 and TRP-2 were determined by real-time PCR.Results Compared with negative control group,after the treatment with butein(1.0 μg·mL~(-1) and 5.0 μg·mL~(-1)) for 48 h,the cell proliferation rate of co-culture system of A375 and Hacat cells were significantly increased(P<0.05 or P<0.01),the content of melanin and TYR activity was increased(P<0.05 or P<0.01),the mRNA expression of TRP-1 and TRP-2 was increased(P<0.01).Conclusion The butein up-regulates TYR activity,as well as the mRNA expression of TRP-1 and TRP-2,and then promotes melanosome synthesis in co-culture system of A375 and Hacat cells.
引文
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