染料木素对A375细胞黑素合成及相关细胞信号通路调控作用的初步探讨

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

摘要:

目的: 研究染料木素对A375细胞黑素合成的影响,并探讨MAPK信号通路与此作用的相关性. 方法: 以人黑素瘤细胞A375为研究对象,将不同浓度的染料木素作用于A375细胞48 h,另设空白组,采用MTT法检测细胞活性;NaOH裂解法检测黑素量;多巴氧化法检测酪氨酸酶(TYR)活性;RT-PCR法测定酪氨酸酶(TYR),酪氨酸酶相关蛋白-1(TRP-1)和酪氨酸酶相关蛋白-2(TRP-2)以及MAPK信号通路关键胞外信号调节激酶1(ERK1),ERK2,c-Jun氨基末端激酶2(JNK2)mRNA的表达. 结果: 与空白组比较,染料木素在安全剂量(10^-1,10^-2,10^-3μmol·L^-1)下均可明显抑制A375细胞黑素合成及TYR活性(P <0.01),1 μmol·L^-1染料木素明显下调A375细胞TYR,TRP-2,TRP-1(P <0.05, P <0.01)mRNA表达,以及ERK1,ERK2,JNK2(P <0.05, P <0.01) mRNA表达. 结论: 染料木素可通过抑制TYR活性和／或下调TYR,TRP-1及TRP-2 mRNA表达,抑制A375细胞的黑素合成,此作用与抑制ERK1,ERK2,JNK2的表达有关.

Abstract:

Objective: To observe the effect of genistein on A375 cells melanogenesis, and to investigate its correlation with mitogen-activated protein kinase (MAPK) signal pathways. Method: A375 was treated by the different concentrations of genistein for 48 h and the blank group was assigned. The cell activity was assayed by using MTT test. The amount of melanin was detected by using NaOH schizolysis method. The activity of tyrosinase (TYR) was detected by using L-dopa oxidation method. The mRNA expressions of tyrosinase (TYR), tyrosinase related proteins-1 (TRP-1), TRP-2, extracellular signal-reg-ulated kinase 1 (ERK1), ERK2 and c-Jun N terminal kinase 2 (JNK2) RT-PCR were measured by using RT-PCR. Result: Compared with control group, genistein at the safe dose (10^-1, 10^-2, 10^-3 μmol·L^-1) could significantly suppress the melanin formation and activity of TYR of A375 cells. genistein at 1 μmol·L^-1 could significantly inhibit the mRNA expressions of TYR, TRP-1, TRP-2, ERK1, ERK2 and JNK2 (P <0.05, P <0.01). Conclusion: It is assumed that genistein could significantly suppress melanogenesis of A375 cells by inhibiting the activity of TYR and down-regulating the expressions of TYR, TRP-1 and TRP-2, the effect may be associated with the expression inhibition of ERK1, ERK2 and JNK2.