紫铆花素上调人黑素瘤细胞A375酪氨酸酶活性及其相关蛋白表达

Butein Upregulates Tyrosinase,Tyrosinase-Related Proteins Expression of A375 Cells

目的探讨驱虫斑鸠菊(Vernonia anthelmintica willd)的黄酮类化合物-紫铆花素对人A375细胞中黑素合成作用的影响。方法采用MTT法、L-Dopa氧化法、NaOH裂解法、Realtime PCR法及Western印迹法分别检测紫铆花素对细胞增殖、黑素含量、酪氨酸酶活性及相关mRNA和蛋白表达的影响。结果在0.5~5.0μg/mL浓度范围内,紫铆花素能够促进人A375细胞内黑素合成和酪氨酸酶(TYR)活性,上调酪氨酸酶相关蛋白(TRP)-1、TRP-2和小眼畸形相关转录因子(MITF)的mRNA和蛋白表达,且无明显细胞毒性。结论紫铆花素可以通过提高酪氨酸酶活性以及促进其相关基因的mRNA和蛋白表达水平,诱导人A375细胞的黑素合成水平。

Objective To evaluate the effect of butein which is a natural compound from Vernonia anthelmintica willd seeds on melanogenesis in A375 cells in vitro.Methods MTT assay was performed to evaluate the proliferative activity,dopa oxidation assay was to evaluate tyrosinase activity,a sodium hydroxide solubilization method was used to measure melanin content,Realtime PCR and Western blot were used to detect mRNA and protein expression.Results The results showed that butein treatment of A375 cells stimulated an increase of intracellular melanin level and tyrosinase(TYR)activity without cytotoxicity within a dose of 0.5~5.0μg/mL.Reverse transcription quantitative polymerase chain reaction and Western blot results also indicated that butein may markedly improve the protein expression and RNA transcription of melanogenic enzyme TYR,microphthalmia-associated transcription factor(MITF),tyrosinase-related protein 1(TRP-1)and tyrosinase-related protein 2(TRP-2).Conclusion Butein can stimulate melanogenesis of A375 cells through up-regulating tyrosinase activity and improving mRNA and protein expression.