白桦脂醇对A375细胞黑素合成及相关细胞信号通路调控机制的研究  

Effect of Betulin on A375 Cells Melanogenesis and Mechanism of Regulating Related Cell Signaling Pathways

作　　者：闫姝[1] 牛春影 徐红丹[1] 熊辉[1] 陈巧云[1] 耿放[2] 张宁[1] 

机构地区：[1]黑龙江中医药大学,哈尔滨150040 [2]哈尔滨师范大学,哈尔滨150025

出　　处：《中国实验方剂学杂志》2016年第10期92-96,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上基金项目(81274035);教育部春晖计划合作科研项目(Z2010018);高校博士点专项基金项目(20112327120009);黑龙江省博士后科研启动金项目(LBH-Q13162);黑龙江中医药大学优秀创新人才支持计划项目(2012)

摘　　要：目的:探讨白桦脂醇治疗黄褐斑的内在作用机制。方法:选择对数生长期的A375细胞,培养24 h,弃掉液体,然后分组并加入不同浓度药液200μL,组别设定为空白组、雌二醇组(1×10^(-3)μmol·L^(-1)浓度的雌二醇)和白桦脂醇组(设定浓度的白桦脂醇),每组均设置6个复孔。用白桦脂醇干预A375细胞,噻唑蓝(MTT)法检测细胞活性;Na OH裂解法检测黑素量;多巴氧化法检测酪氨酸酶(TYR)活性;逆转录-聚合酶链式反应(RT-PCR)法检测TYR,酪氨酸酶相关蛋白-1(TRP-1)和酪氨酸酶相关蛋白-2(TRP-2)的表达,及丝裂原激活的蛋白激酶(MAPK)信号通路中关键蛋白激酶ERK1,ERK2,JNK2的mRNA表达。结果:与空白组比较,白桦脂醇在1,1×10^(-1),1×10^(-2),1×10^(-3)μmol·L^(-1)剂量下可明显抑制A375细胞黑素合成及TYR活性,1μmol·L^(-1)白桦脂醇可明显下调A375细胞TYR,TRP-1,TRP-2 mRNA表达,以及ERK1,ERK2,JNK2 mRNA表达(P<0.05,P<0.01)。结论:白桦脂醇可能是通过抑制TYR活性和/或下调TYR,TRP-1及TRP-2 mRNA表达,抑制了A375细胞中黑素的合成,并且这种作用与抑制ERK1,ERK2,JNK2的基因表达有关。Objective： To investigate the potential mechanism of betulin in treatment of chloasma.Method： The A375 cell lines in logarithmic phase were cultured for 24 hours,and then the liquid was discarded.The cultured cells were divided and added with different concentrations of 200 μL drug liquid,and then divided into blank group,estradiol group（ 1 × 10^-3μmol·L^-1estradiol）,betulin group（ at set concentration）,with 6wells in each group. A375 cells were treated with betulin and then methyl thiazdyl tetrazolium（ MTT） assay was used to detect cells activity; Na OH schizolysis method and L-dopa oxidation method were respectively used to detect amount of melanin and the activity of tyrosinase（ TYR）. Reverse transcription-polymerase chain reaction（ RT-PCR） was applied to measure the expression levels of tyrosinase（ TYR）,tyrosinase related protein-（-1）（ TRP-1） and tyrosinase related protein-2（ TRP-2）,as well as the mRNA expression levels of the key protein kinases ERK1,ERK2 and JNK2 in MAPK signal pathways. Result： As compared with the blank group,betulin of 1,1 × 10^-1,1 × 10^-2,1 × 10^-3μmol·L^-1can significantly inhibit melanin formation of A375 cells and activity of tyrosinase（ TYR）; betulin of 1 μmol ·L^-1 can significantly down-regulate mRNA expression levels of TYR,TRP-1,TRP-2 as well as ERK1,ERK2 and JNK2（ P 〈0. 05,P〈0. 01）. Conclusion： Betulin can inhibit the melanogenesis of A375 cells probably by inhibiting TYR activity and/or down-regulating TYR,TRP-1 and TRP-2mRNA expression levels,and this effect may be associated with inhibition of the expression of ERK1,ERK2 and JNK2 levels.

关 键 词：黄褐斑:白桦脂醇 A375细胞 黑素 MAPK信号通路 

分 类 号：R285.5[医药卫生—中药学]