SIRT1基因激活后鸡巨噬细胞转录组分析

doi:10.11843/j.issn.0366-6964.2025.06.012

摘要/Abstract

摘要：

旨在激活SIRT1，通过转录组测序探究其在家禽巨噬细胞中的免疫调节作用及相关靶基因和信号通路。本试验用SIRT1的一种选择性激活剂：SRT1720 HCl 1 μmol·L^-1处理鸡巨噬细胞，试验分为对照组(DMSO)和试验组(SRT1720 HCl)，每组5个生物学重复，处理24 h后通过提取细胞总RNA反转录，使用Illumina Novaseq X Plus进行高通量测序，利用生物信息学方法进行分析。以|log₂FC|>1和校正后的P < 0.05为阈值筛选差异表达基因，共筛选到1 183个差异表达表达基因(P < 0.05)，其中781个基因表达上调，402个基因表达下调，包括CD14、TLR7、IL10RA、NFKBIA和JUN等重要基因。从差异表达基因中选取5个基因进行实时荧光定量验证，结果与转录组结果一致。对筛选到的差异表达基因进行GO和KEGG功能富集分析，GO分析显著富集到代谢过程、免疫系统过程、生物粘附和催化活性等GO条目(P < 0.05)；KEGG分析显著富集到MAPK信号通路、TOLL样受体信号通路和NOD样受体信号通路等，值得注意的是分析富集到了沙门氏菌感染信号通路，提示激活禽巨噬细胞SIRT1后可能影响感染沙门氏菌；通过构建蛋白互作网络发现SIRT1可能与NFKBIA、JUN等基因关系密切。本研究利用SIRT1基因激活后鸡巨噬细胞转录组分析筛选到与免疫相关的基因和重要通路，为探究SIRT1在家禽免疫中的作用提供研究基础。

关键词: SIRT1, 禽巨噬细胞, 转录组

Abstract:

This study aimed to activate avian macrophages using a selective SIRT1 activator and identify relevant target genes and signaling pathways through transcriptomic analysis. Chicken macrophages were treated with a selective SIRT1 activator, SRT1720 HCl, at a concentration of 1 μmol·L^-1. The experiment was divided into a control group (treated with DMSO) and an experimental group (treated with SRT1720 HCl), with 5 biological replicates per group. After 24 hours of treatment, total cellular RNA was extracted and reverse transcribed. High-throughput sequencing was then performed using the Illumina Novaseq X Plus platform, and subsequent bioinformatics analyses were conducted. Differentially expressed genes (DEGs) were identified using thresholds of |log₂ FC|>1 and an adjusted P < 0.05. A total of 1 183 DEGs were detected (P < 0.05), of which 781 were upregulated and 402 were downregulated, including key genes such as CD14, TLR7, IL10RA, NFKBIA, and JUN. Five genes selected from the DEGs were validated by real-time quantitative PCR, and the results were consistent with the transcriptomic data. Gene Ontology (GO) enrichment analysis revealed significant enriched terms associated with metabolic processes, immune system processes, biological adhesion, and catalytic activity (P < 0.05). Additionally, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis results showed significant enriched pathways including the MAPK signaling pathway, Toll-like receptor signaling pathway, and NOD-like receptor signaling pathway. Notably, enrichment of the Salmonella infection pathway was observed, suggesting that activation of SIRT1 in avian macrophages might influence Salmonella infection. Protein-protein interaction network analysis further indicated that SIRT1 was closely related to genes such as NFKBIA and JUN. The study utilized transcriptome analysis of chicken macrophages after SIRT1 gene activation to screen for immune-related genes and important pathways, provides a research foundation for exploring the role of SIRT1 in avian immunity.

Key words: SIRT1, avian macrophage, transcriptome

中图分类号:

S831.2

刘莎, 苏蒙, 高倩梅, 宋丹丽, 赵桂苹, 李建慧, 李庆贺. SIRT1基因激活后鸡巨噬细胞转录组分析[J]. 畜牧兽医学报, 2025, 56(6): 2661-2671.

LIU Sha, SU Meng, GAO Qianmei, SONG Danli, ZHAO Guiping, LI Jianhui, LI Qinghe. Transcriptome Analysis of Chicken Macrophages after SIRT1 Activated[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(6): 2661-2671.

图/表 5

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基因Gene	上游引物(5′→3′)Forward	下游引物(5′→3′)Reverse
β-actin	GAGAAATTGTGCGTGACATCA	CCTGAACCTCTCATTGCCA
CD14	ACCCGACACTTGACCCTGTTG	TCTGCCTTAGGGGAAAAGGGC
NFKBIA	TGGGAGAACAGCACTACACT	GCCCTGGTAGGTCACTTTG
TLR7	GCACACCGGAAAATGGTACA	TTGGGAAACCAACGTCCTGAT
JUN	AACTCCGCACCCAACTAC	GGTACAGTCTGAGGCTCTTCT
IL10RA	TGGGCATCTTCGACACTGAC	GAGTTGGTTTGCACCGTGAG

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