阿魏酸调节C-KIT/MITF蛋白表达影响黑素生成的实验研究

摘要

目的 探讨不同浓度阿魏酸对UVB诱导的A375细胞黑素生成的影响及其作用机制.方法 常规体外培养A375细胞,分为空白组、模型组、阿魏酸低浓度(10 μmol/L)组、阿魏酸高浓度(100 μmol/L)组和阳性对照组,除空白组外,各组均以UVB 20 mJ/cm2照射,阿魏酸高低浓度组加含阿魏酸的培养液、阳性对照组加入含熊果苷的培养液作用24h,四甲基偶氮唑蓝(MTT)比色法测定细胞增殖;采用多巴氧化法研究酪氨酸酶活性的变化;比色法测定黑素含量,免疫印迹法测定C-KIT和MITF蛋白表达.结果 与空白组比较,模型组细胞酪氨酸酶活性和黑素含量明显增加,阿魏酸低、高浓度组和阳性对照组均可抑制UVB诱导的A375细胞酪氨酸酶活性增高和黑素生成,其机制可能是通过下调C-KIT和MITF蛋白表达而实现的.结论 高浓度阿魏酸可明显下调C-KIT/MITF蛋白表达,进而抑制酪氨酸酶活性和黑素生成.%Objective To investigate the effect of the different concentrations of ferulic acid on the melanin fonnation of UVB induced A375 cells and its mechanism.Methods A375 cells cultivated in vitro were applied in this study,and they were divided into the control group,model group,low concentration of ferulic acid (10 μmol/L) group,high concentration of ferulic acid (100 μmol/L) group,and positive control group.Except for the control group,all other groups were treated with 20 mJ/cm2.Low and high concentration of ferulic acid groups were treated with culture medium containing ferulic acid for 24 h,while the positive control group was treated with arbutin for 24 h,MTF method was used to detect the cell proliferation;dopamine oxidation method was used to analysis the variation of tyrosinase activity;the melanin content was observed with colorimetric method;the proteins expression of C-KIT and MITF were observed by Western blot.Results The tyrosinase activity and melanin formation significantly increased in model group compared with the control group,low,high concentration ferulic acid inhibited the tyrosinase activity and melanin formation of UVB induced A375 cells,which might be achieved by lowering the proteins expression of C-KIT and MITF.Conclusion High concentration of ferulic acid significantly down-regulated the proteins expression of C-KIT/MITF,furthermore inhibited tyrosinase activity and melanin formation.