补骨脂素对A375细胞黑素合成及ER/MAPK信号通路调控机制探讨

崔悦; 单孟瑶; 张丽宏; 刘国良; 董蕊; 赵旭; 张宁; 耿放

doi:10.13422/j.cnki.syfjx.20181932

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补骨脂素对A375细胞黑素合成及ER/MAPK信号通路调控机制探讨

药理 | 更新时间：2024-01-04

- 补骨脂素对A375细胞黑素合成及ER/MAPK信号通路调控机制探讨
- Mechanism of Psoralen on Melanin Synthesis and ER/MAPK Signaling Pathway in A375 Cells
- 中国实验方剂学杂志 2018年24卷第19期页码：153-158
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金面上项目（81274035，81673621）
- DOI：10.13422/j.cnki.syfjx.20181932
  中图分类号： R285.5
- 纸质出版日期：2018
- 稿件说明：
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崔悦, 单孟瑶, 张丽宏, 等. 补骨脂素对A375细胞黑素合成及ER/MAPK信号通路调控机制探讨[J]. 中国实验方剂学杂志, 2018,24(19):153-158.

CUI Yue, SHAN Meng-yao, ZHANG Li-hong, et al. Mechanism of Psoralen on Melanin Synthesis and ER/MAPK Signaling Pathway in A375 Cells[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(19): 153-158.
崔悦, 单孟瑶, 张丽宏, 等. 补骨脂素对A375细胞黑素合成及ER/MAPK信号通路调控机制探讨[J]. 中国实验方剂学杂志, 2018,24(19):153-158. DOI： 10.13422/j.cnki.syfjx.20181932.

CUI Yue, SHAN Meng-yao, ZHANG Li-hong, et al. Mechanism of Psoralen on Melanin Synthesis and ER/MAPK Signaling Pathway in A375 Cells[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(19): 153-158. DOI： 10.13422/j.cnki.syfjx.20181932.

摘要

目的：探讨补骨脂素对A375细胞黑素含量、酪氨酸酶活性的影响及作用机制。方法：以密度2×105个/孔的细胞数接种于6孔板中，将细胞分为空白组、雌二醇组、补骨脂素组、补骨脂素+雌激素受体（ER）阻断剂组（ICI182780）（浓度分别为1×10-3，1，1 μmol·L-1）。采用NaOH裂解法、多巴醌氧化法检测黑素含量及酪氨酸酶活性；蛋白免疫印迹法（Western blot）和实时荧光定量聚合酶链式反应（Real-time PCR）法测定ER/丝裂原激活的蛋白激酶（ER/MAPK）通路中关键蛋白激酶p38，c-Jun氨基末端激酶（JNK），细胞外信号调节激酶2（ERK2）和小眼畸形相关转录因子（MITF），多巴氧化法检测酪氨酸酶（TYR），酪氨酸酶相关蛋白-1（TRP-1），酪氨酸酶相关蛋白-2（TRP-2）的蛋白含量及相应mRNA表达。结果：与空白组比较，补骨脂素组对A375细胞中的黑素合成及TYR活性具有显著抑制作用（P<0.01），对A375细胞中雌激素受体β（ERβ）蛋白的表达具有明显的抑制作用（P<0.05），对p38 MAPK，ERK2，JNK的蛋白表达极显著的抑制作用（P<0.01），对MITF，TYR，TRP-1，TRP-2的蛋白表达极显著的抑制作用（P<0.01），对ERK2，p38 MAPK，MITF，TRP-1 mRNA表达显著被抑制（P<0.01）；与补骨脂素组比较，ICI182780能够逆转ERβ，ERK，MITF，TYR，TRP-1的蛋白表达（P<0.01）及p38MAPK，JNK，TRP-2的蛋白表达（P<0.05），同时能显著阻断补骨脂素对ERK2，p38MAPK，MITF，TRP-1 mRNA表达作用（P<0.01）。结论：补骨脂素通过ER/MAPK信号通路下调MITF和TYR，TRP-1，TRP-2的表达量，从而减少黑素合成。

Abstract

Objective: To investigate the effect and mechanism of psoralen on melanin content and tyrosinase activity in A375 cells. Method: Cells were inoculated in 6-well plates at a cell density of 2×105 cells and then divided into blank group

estradiol group

psoralen group

psoralen+estrogen receptor(ER) receptor blocker group (ICI182780)

with the concentrations of 1×10-3

1 μmol·L-1

respectively. The melanin content and tyrosinase activity were detected by NaOH lysis method and dopaquinone oxidation method; Western blot and real-time fluorescent quantitative PCR assays were used to determine key protein kinases p38

c-Jun N-terminal kinase(JNK)

extracellular signal-regulated kinase 2 (ERK2) and ocular hypotrophy-associated transcription factors (MITF) in the ER/mitogen-activated protein kinase(MAPK) pathway. The protein contents and corresponding mRNA expressions of tyrosinase (TYR)

tyrosinase-associated protein-1 (TRP-1)

and tyrosinase-associated protein-2 (TRP-2) were measured by dopa oxidation. Result: Compared with the blank group

the psoralen group had a significant inhibitory effect on melanin synthesis and TYR activity in A375 cells(P<0.01)

with a significant inhibitory effect on the expression of estrogen receptor β(ERβ) protein in A375 cells (P<0.05)

p38 MAPK

ERK and JNK protein expressions (P<0.01)

protein expressions of MITF

TYR

TRP-1

and TRP-2 (P<0.01)

and ERK2

p38 MAPK

MITF and TRP-1 mRNA expressions (P<0.01); compared with psoralen group

ICI182780 reversed the protein expressions of ERβ

ERK

MITF

TYR

TRP-1 (P<0.01) and the protein expressions of p38

JNK

and TRP-2 (P<0.05); at the same time

it can significantly block psoralen on ERK2

p38

MITF

TRP-1 mRNA expressions (P<0.01). Conclusion: Psoralen down-regulates the expressions of MITF and TYR

TRP-1

and TRP-2 through ER/MAPK signaling pathway

so as to reduce melanin synthesis.

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