摘要
目的探讨葛根素对正常人黑素细胞黑素合成的影响和可能机制。方法用噻唑蓝（MTr）法、NaOH裂解法观察葛根素对黑素细胞增殖及黑素合成作用，RT—PCR及Western印迹法检测葛根素对黑素细胞小眼畸形相关转录因子（MITF）、酪氨酸酶（rIYR）、酪氨酸酶相关蛋白1（TRP-1）基因转录和蛋白表达水平的影响。结果1—40μmol／L浓度范围内葛根素对体外培养的黑素细胞增殖影响与正常对照组相比，差异无统计学意义（P〉0．05）。与正常对照组相比，40μmol／L葛根素可显著促进黑素细胞的黑素合成（P〈0．05），并可显著增加MITF、TYR、TRP-1mRNA及蛋白的表达（P〈0．05）。40μmol／L葛根素使MITF、TYR、TRP-1的蛋白表达量分别比正常对照组增加8．69％，10．28％和10．58％（P〈O．05）；并使MITF、TYR、TRP-1的mRNA表达量分别比正常对照组增加2．48倍，1．91倍和1．63倍（P〈0．05）。结论葛根素能增加MITF、TYR、TRP-1mRNA及蛋白的表达水平，促进黑素合成。
Objective To evaluate the effect of puerarin on melanogenesis in melanocytes, and to explore its possible mechanisms. Methods Third- to fifth-passage melanocytes isolated from human foreskin were treated with different concentrations （ 1, 5, 10, 20, 40, 80 and 160 μmol/L） of puerarin for 24 hours, with those receiving no treatment as the normal control group. Methyl thiazolyl tetrazolium （MTT） assay was performed to evaluate the proliferative activity of melanocytes, a sodium hydroxide solubilization method was used to measure melanin content, and reverse transcription PCR （RT-PCR） and Western blot analysis were performed to quantify the mRNA and protein expressions of microphthalmia-assoeiated transcription factor （MITF）, tyrosinase （TYR） and tyrosinase-related protein-1 （TRP-1） respectively. Results There were no significant differences in the proliferative activity of melanoeytes between the puerarin （ 1 - 40 p, mol/L） groups and normal control group （P 〉 0.05 ）, and 40 μmol/L was chosen as the concentration of puerarin for subsequent experiments. Compared with the normal control group, the 40-μmol/L puerarin group showed increased melanin content as well as mRNA and protein expressions of MITF, TYR and TRP-1 （all P 〈 0.05 ）. Concretely speaking, the protein expressions of MITF, TYR and TRP-1 in the 40-p, mol/L puerarin group were increased by 8.69%, 10.28% and 10.58% compared with the normal control group respectively （all P 〈 0.05）, and their mRNA expressions were 2.48, 1.91 and 1.63 times higher in the 40-p, mol/L puerarin group than in the normal control group respectively （all P 〈 0.05）. Conclusion Puerarin can increase the mRNA and protein expressions of MTTF, TYR and TRP-1, and promote melanogenesis in melanoeytes.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2016年第5期338-341,共4页
Chinese Journal of Dermatology
基金
国家自然科学基金（81271758、81472887）
浙江省医药卫生重大科技计划项目（WKJ2012-2-306）
关键词
白癜风
黑素细胞
葛根素
细胞增殖
小眼畸形相关转录因子
一元酚单氧酶
Vitiligo
Melanoeytes
Puerarin
Cell proliferation
Mierophthalmia-associated transcription factor
Monophenol monooxygenase
作者简介
通信作者：许爱娥，Email：xuaiehz@msn．com
