ASIP-YY对羊驼皮肤黑色素细胞形态和黑色素生成的影响

发布时间：2018-03-17 04:15

本文选题：ASIP-YY　切入点：羊驼　出处：《农业生物技术学报》2017年07期 　论文类型：期刊论文

【摘要】：鼠灰色(Agouti)基因时影响动物毛色的重要基因之一,虽然在多种动物中发现了Agouti基因的存在和表达,但Agouti信号蛋白(Agouti signaling protein,ASIP)对羊驼(Lama pacos)黑色素细胞的增殖及对毛色相关基因的影响研究鲜见报道。本研究旨在利用不同浓度ASIP-YY(a C-terminal fragment of ASIP)作用于体外培养的羊驼皮肤黑色素细胞,观察其对黑色素细胞及黑色素生成的影响。不同浓度ASIP-YY(0.0、0.1、1.0、10.0、100.0 nmol/L)处理羊驼皮肤黑色素细胞72 h。四甲基偶氮噻唑蓝(methyl thiazolyl tetrazolium,MTT)法检测黑色素细胞活力,紫外分光光度法检测黑色素含量,q RT-PCR检测酪氨酸酶(Tyrosinase,TYR)、TRP-1、TRP-2、黑色素细胞膜上的黑色素皮质激素受体1(melanocortin-1 receptor,MC1R)和小眼畸形相关转录因子(Micropht-halmiaassocitated transcription factor,MITF)m RNA表达量。结果显示,黑色素细胞活力受到抑制;ASIP-YY在0.1、10.0、100.0 nmol/L时对TYR m RNA的表达极显著抑制(P0.01),10.0、100.0 nmol/L时对TRP-1 m RNA的表达极显著抑制(P0.01),10.0 nmol/L时对MC1R m RNA的表达显著抑制(P0.05),0.1、1.0、10.0 nmol/L时对MITF m RNA的表达极显著抑制(P0.01);0.1、1.0 nmol/L时对TRP-1、TRP-2 m RNA的表达显著提高(P0.05),1.0、100.0 nmol/L时对MC1R的表达极显著提高(P0.01)。ASIP-YY可使黑色素细胞体积变小,抑制黑色素相关基因的表达,可能是ASIPYY竞争性的与MC1R结合,从而抑制黑色素相关基因的表达。本实验为进一步研究ASIP对羊驼皮肤黑色素生成调控提供一定的理论基础。
[Abstract]:The Agouti gene is one of the important genes that influence the color of animal coat, although the existence and expression of Agouti gene have been found in many animals. However, there are few reports on the effects of Agouti signal protein (Agouti) on the proliferation of melanocytes and on the coat color related genes. The aim of this study was to use different concentrations of ASIP-YY(a C-terminal fragment of ASIP to treat the melanocytes of alpaca skin in vitro. To observe its effect on melanocyte and melanin production, the melanocytes of alpaca skin were treated with different concentrations of ASIP-YYP0.00.1nmol / L (1.0 ~ 10.0nmol / L) for 72 h. The activity of melanocytes was detected by tetrazoliumium tetrazolium tetrazolium tetrazolium (MTT) assay. Detection of melanin content by UV Spectrophotometry and RT-PCR Detection of tyrosinase TRP-1 TRP-2, melanocortin-1 receptor MC1R on melanin cell membrane and micropht-halmiaassocitated transcription factorMITFm RNA expression on melanocortin-1 receptor on melanoma cell membrane and micropht-halmiaassocatted transcription factorMITFm RNA, the results showed that. The expression of TYR m RNA was significantly inhibited by ASIP-YY at 0.1 ~ 10.0 ~ 100.0 nmol/L. The expression of TRP-1 m RNA was significantly inhibited by TRP-1 m RNA at 0.01 ~ 10.0 nmol/L. ASIP-YY significantly inhibited the expression of MC1R m RNA at 10. 0 nmol/L with P0. 05 0. 1. 01 and 10. 0 nmol/L. The expression of MITF m RNA was significantly inhibited by P0. 01. 0. 0. 0. 0. 0 nmol/L and 10. 0% P0. 01. 0. 0. 0. 0. 1. 0 nmol/L. The expression of TRP-1 m RNA was significantly inhibited by ASIP-YY. Inhibition of the expression of TRP-1TRP-2 m RNA at 1.0 nmol/L of P0.01P0.01P0.01TP-YY could significantly increase the expression of MC1R at 1.0 nmol/L. ASIP-YY could make the melanocyte volume smaller. Inhibiting the expression of melanin related genes may be the competitive binding of ASIPYY to MC1R, thus inhibiting the expression of melanin related genes. This study provides a theoretical basis for further study on the regulation of melanin production in alpaca skin by ASIP.
【作者单位】：山西农业大学动物科技学院;
【基金】：国家自然科学基金项目(No.30972223和No.31272628)
【分类号】：S829.9

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