结合 RNA-seq 揭示褪黑素在丝羽乌骨鸡原代黑素细胞中黑色素合成中的作用。

Combine with RNA-seq Reveals the Effect of Melatonin in the Synthesis of Melanin in Primary Melanocytes of Silky Fowls Black-Bone Chicken.

机构信息

Key Laboratory for Animal Genetics & Molecular Breeding of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.

Institute of Epigenetics and Epigenomics, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.

出版信息

Genes (Basel). 2023 Aug 18;14(8):1648. doi: 10.3390/genes14081648.

DOI:10.3390/genes14081648

PMID:37628699

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10454632/

Abstract

(1) Background: It was found that the melanin of black-bone chicken has various effects such as scavenging DPPH free radicals and anti-oxidation, and the synthesis of melanin is affected by various factors including hormones. In addition, several studies have found that melatonin affects the melanoma cell synthesis of melanin, which has not been reported in chicken primary melanocytes; so, relevant studies were conducted. (2) Methods: In this study, chicken primary melanocytes were isolated and characterized, and then melanocytes were treated with different concentrations of melatonin to investigate the effects of melatonin on melanin synthesis in chicken melanocytes in terms of melanin synthesis-related genes, melanin content, and tyrosinase activity, and combined with RNA seq to detect the change in gene expression level of chicken melanocytes after melatonin treatment. (3) Results: We isolated and characterized primary melanocytes, and indirect immunofluorescence assay results showed positive melanocyte marker genes. RT-qPCR results showed that melatonin decreased the expression of melanin synthesis-related genes. In addition, melatonin reduced the melanin content and decreased the tyrosinase activity of melanocytes in the treated group. A total of 1703 differentially expressed genes were screened by RNA-seq, and in addition, in the KEGG results, the signaling pathway associated with melanin synthesis, and the mTOR signaling pathway were enriched. (4) Conclusions: Melatonin could decrease the synthesis of melanin in chicken primary melanocytes.

摘要

(1) 背景：研究发现黑骨鸡的黑色素具有清除 DPPH 自由基和抗氧化等多种作用，而黑色素的合成受激素等多种因素的影响。此外，有几项研究发现褪黑素影响黑色素瘤细胞黑色素的合成，但在鸡原代黑素细胞中尚未有报道；因此，进行了相关研究。(2) 方法：本研究分离并鉴定了鸡原代黑素细胞，然后用不同浓度的褪黑素处理黑素细胞，从黑色素合成相关基因、黑色素含量和酪氨酸酶活性等方面探讨褪黑素对鸡黑素细胞黑色素合成的影响，并结合 RNA seq 检测褪黑素处理后鸡黑素细胞基因表达水平的变化。(3) 结果：我们分离并鉴定了原代黑素细胞，间接免疫荧光检测结果显示阳性的黑素细胞标记基因。RT-qPCR 结果表明褪黑素降低了黑色素合成相关基因的表达。此外，褪黑素降低了处理组黑素细胞的黑色素含量和酪氨酸酶活性。通过 RNA-seq 共筛选出 1703 个差异表达基因，此外，KEGG 结果富集到与黑色素合成相关的信号通路和 mTOR 信号通路。(4) 结论：褪黑素可以降低鸡原代黑素细胞中黑色素的合成。