小鼠黑色素细胞沉默及过表达色素上皮衍生因子对黑色素合成的影响

doi:10.11843/j.issn.0366-6964.2023.09.030

摘要/Abstract

摘要： 旨在研究色素上皮衍生因子(pigment epithelium-derived factor, PEDF)与黑色素合成调控因子之间的互作关系,探索PEDF对黑色素细胞活力和黑色素合成的调控作用,为研究动物毛色形成机制提供基础。本研究将PEDF siRNA及PEDF过表达载体分别转染小鼠黑色素细胞,对黑色素细胞活力、黑色素含量进行检测,采用qRT-PCR、Western blotting及细胞免疫荧光对PEDF、Wnt1、Wnt3α、MITF和β-catenin的表达和定位进行检测。黑色素细胞PEDF沉默后,黑色素细胞的细胞活力增强,黑色素含量增加,Wnt1、Wnt3α、MITF和β-catenin的mRNA和蛋白表达量升高;黑色素细胞PEDF过表达后,黑色素细胞活力减弱,黑色素含量减少,Wnt1、Wnt3α、MITF和β-catenin的mRNA和蛋白表达量降低。细胞免疫荧光结果显示:PEDF、Wnt1、MITF、β-catenin表达于黑色素细胞的细胞质;Wnt3α表达于黑色素细胞的细胞质和细胞膜。本研究证实:PEDF使黑色素细胞活力减弱,可通过下调促黑色素生成相关的因子Wnt1、Wnt3α、MITF和β-catenin,从而抑制黑色素的生成。

关键词: 色素上皮衍生因子, PEDF, 沉默和过表达, 黑色素, 小鼠黑色素细胞

Abstract: The purpose of this study was to investigate the interaction between pigment epithelium-derived factor (PEDF) and melanin synthesis regulatory factors, and to explore the regulatory effects of PEDF on melanin cell vitality and melanin synthesis, so as to provide a basis for the study of animal coat color formation mechanism. In this study, PEDF siRNA and PEDF overexpression vector were transfected into mouse melanocytes respectively, and the activity of melanocytes and content of melanin were detected. The expression and localization of PEDF, Wnt1, Wnt3α, MITF and β-catenin were detected by qRT-PCR, Western blotting and cellular immunofluorescence. After PEDF silencing, the cell vitality of melanocytes was enhanced, the content of melanin was increased, and the mRNA and protein expressions of Wnt1, Wnt3α, MITF and β-catenin were increased. After PEDF overexpression in melanocytes, the activity of melanocytes was weakened, the content of melanin was decreased, and the mRNA and protein expressions of Wnt1, Wnt3α, MITF and β-catenin were decreased. Cellular immunofluorescence results showed that PEDF, Wnt1, MITF and β-catenin were expressed in the cytoplasm of melanocytes. Wnt3α was expressed in the cytoplasm and cell membrane of melanocytes. This study could confirm that PEDF could reduce the activity of melanocytes, and inhibit the production of melanin by down-regulating the factors related to melanin production, Wnt1, Wnt3α, MITF and β-catenin.

Key words: pigment epithelium-derived factor, PEDF, silence and overexpression, melanin, mouse melanocyte

中图分类号:

Q342

梁睿, 范小瑞, 张晋强, 庞全海. 小鼠黑色素细胞沉默及过表达色素上皮衍生因子对黑色素合成的影响[J]. 畜牧兽医学报, 2023, 54(9): 3916-3930.

LIANG Rui, FAN Xiaorui, ZHANG Jinqiang, PANG Quanhai. Effects of Mouse Melanocyte Silencing and Overexpression of Pigment Epithelium-Derived Factor on Melanin Synthesis[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3916-3930.

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