ZBED6基因敲除对巴马香猪肺脏转录调控的影响

doi:10.16431/j.cnki.1671-7236.2025.12.029

摘要/Abstract

摘要： 【目的】研究ZBED6基因敲除对巴马香猪肺脏基因表达谱的影响，进一步明确ZBED6基因缺失对巴马香猪肺脏功能的影响。【方法】采集8月龄雌性野生型(WT)和ZBED6基因敲除型(ZBED6-KO)巴马香猪肺脏组织并称重，用Trizol法提取肺脏组织RNA，经质量检测后建库(NEBNext^® Ultra^TM)并测序(Illumina HiSeq 2500)。数据经Fastp v 0.23.4质控，TopHat和Cufflink分析差异基因，以|log₂FoldChange|≥1和P≤0.05为标准筛选出差异表达基因。利用g:Profiler在线数据库对差异表达基因进行富集分析，利用实时荧光定量PCR对转录组测序(RNA-Seq)结果进行验证。【结果】与WT型巴马香猪相比，ZBED6-KO型巴马香猪的肺脏重量无显著变化(P＞0.05)。利用转录组测序在WT和ZBED6-KO型巴马香猪肺脏组织中共筛选出480个差异表达基因，其中上调基因125个，下调基因355个。上调基因主要富集在2条与免疫相关的通路，下调基因主要富集在代谢相关通路中。实时荧光定量PCR验证结果显示，上调和下调基因的表达变化趋势与RNA-Seq结果一致，证实了RNA-Seq数据的可靠性。【结论】ZBED6基因敲除影响了巴马香猪肺脏组织的基因表达谱，其中免疫相关基因普遍上调，而代谢相关基因显著下调。本研究结果为ZBED6在肺脏组织中的功能与作用提供更多的理论支持和证据，进一步填补了转录因子ZBED6在哺乳动物中肺脏方面作用机制的研究空白。

关键词: ZBED6基因; 基因敲除; 转录组分析; 肺脏; 巴马香猪

Abstract: 【Objective】 This study aimed to investigate the effects of ZBED6 gene knockout on the gene expression profile in lungs of Bama Xiang pigs and further elucidate the impact of ZBED6 deficiency on pulmonary function in this breed. 【Method】 Lung tissues were collected from 8-month-old female wild-type (WT) and ZBED6-knockout (ZBED6-KO) Bama Xiang pigs,and lung weight was measured.Total RNA was extracted using TRIzol reagent,followed by quality assessment,library preparation (NEBNext^® Ultra^TM),and sequencing (Illumina HiSeq 2500).Raw data were quality-controlled using Fastp v 0.23.4,and differentially expressed genes (DEGs) were analyzed using TopHat and Cufflink,with the threshold set at |log₂FoldChange|≥1 and P≤0.05.Functional enrichment analysis of DEGs was performed using the g:Profiler online database,and RNA-Seq results were validated by Real-time quantitative PCR. 【Result】 Compared with WT Bama Xiang pigs,the lung weight of ZBED6-KO Bama Xiang pigs showed no significant changes (P＞0.05).RNA sequencing (RNA-Seq) analysis identified 480 DEGs between WT and ZBED6-KO lungs of Bama Xiang pigs,including 125 upregulated and 355 downregulated genes.Upregulated genes were primarily enriched in two immune-related pathways,while downregulated genes were significantly associated with metabolic pathways.Real-time quantitative PCR validation confirmed that the expression trends of selected DEGs were consistent with the RNA-Seq results,confirming the reliability of the sequencing data. 【Conclusion】 ZBED6 gene knockout significantly altered the gene expression profile in lungs of Bama Xiang pigs,with immune-related genes generally upregulated and metabolic-related genes downregulated.This study provided theoretical basis for understanding the role of ZBED6 gene in pulmonary function,further filling the research gap regarding the regulatory mechanisms of the transcription factor ZBED6 in mammalian lungs.

Key words: ZBED6 gene; gene knockout; transcriptome analysis; lung; Bama Xiang pigs

中图分类号:

S828

陈欣雨, 王圣楠, 王丹丹, 马月辉, 蒋琳, 刘书琴. ZBED6基因敲除对巴马香猪肺脏转录调控的影响[J]. 中国畜牧兽医, 2025, 52(12): 5849-5858.

CHEN Xinyu, WANG Shengnan, WANG Dandan, MA Yuehui, JIANG Lin, LIU Shuqin. Effects of ZBED6 Gene Knockout on Transcriptional Regulation in Lungs of Bama Xiang Pigs[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(12): 5849-5858.

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