转录因子CEBPB对猪δ冠状病毒体外复制的抑制作用

doi:10.11843/j.issn.0366-6964.2025.07.034

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (7): 3423-3432.doi: 10.11843/j.issn.0366-6964.2025.07.034

转录因子CEBPB对猪δ冠状病毒体外复制的抑制作用

陈琢琦¹^,²(), 范丽原², 钟纯燕⁵, 于岩飞², 李基棕²^,³^,⁴^,*(), 李彬²^,³^,⁴^,*(), 袁晓民¹^,*()

1. 湖南农业大学动物医学院，长沙 410128
2. 江苏省农业科学院兽医研究所农业部兽用生物制品工程技术重点实验室，南京 210014
3. 江苏大学生命科学学院，镇江 212013
4. 南京农业大学动物医学院，南京 210095
5. 黔西南民族职业技术学院生物工程系，兴义 562400

收稿日期:2024-07-05 出版日期:2025-07-23 发布日期:2025-07-25
通讯作者: 李基棕,李彬,袁晓民 E-mail:475846734@qq.com;lijizong22@sina.com;libinana@126.com;yxm1230@hunau.edu.cn
作者简介:陈琢琦(2000-)，男，河南三门峡人，硕士生，主要从事动物传染病防治和诊断技术研究，E-mail: 475846734@qq.com
基金资助:
“十四五”国家重点研发计划(2023YFD1801303)；国家自然科学基金(32473119)；江苏省自然科学基金(BK20221432)；湖南省教育厅科学研究项目(22B0224)；云南省科技重大专项(202202AE 090032)；农业农村部兽医生物制品工程技术重点实验室开放课题(SYKF(23)002)

The Inhibitory Effect of Transcription Factor CEBPB on the Replication of Porcine Deltacoronavirus in vitro

CHEN Zhuoqi¹^,²(), FAN Liyuan², ZHONG Chunyan⁵, YU Yanfei², LI Jizong²^,³^,⁴^,*(), LI Bin²^,³^,⁴^,*(), YUAN Xiaomin¹^,*()

1. College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China
2. Key Laboratory of Engineering Technology for Animal Biological Products, Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
3. College of Life Science, Jiangsu University, Zhenjiang 212013, China
4. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
5. Department of Bioengineering, Southwest Guizhou Vocational and Technical College for Nationalities, Xingyi, 562400, China

Received:2024-07-05 Online:2025-07-23 Published:2025-07-25
Contact: LI Jizong, LI Bin, YUAN Xiaomin E-mail:475846734@qq.com;lijizong22@sina.com;libinana@126.com;yxm1230@hunau.edu.cn

摘要/Abstract

摘要：

本研究旨在探索CCAAT增强子结合蛋白β(CEBPB)对猪δ冠状病毒(porcine deltacoronavirus, PDCoV)复制的影响。将PDCoV接种于LLC-PK1细胞，检测不同时间点CEBPB表达水平变化；过表达和敲低CEBPB，利用qPCR和Western blot评估其对PDCoV复制的影响；并进一步明确CEBPB在PDCoV复制周期中的作用；利用qPCR筛选出影响CEBPB上调的病毒蛋白，共聚焦显微镜观察PDCoV病毒蛋白与CEBPB的共定位现象。结果显示，随着病毒感染时间的延长，细胞中CEBPB的表达量呈上升趋势，过表达CEBPB显著抑制了PDCoV的复制，而敲低CEBPB则促进病毒的复制，CEBPB主要作用于PDCoV的复制阶段，并且PDCoV Nsp4能促进CEBPB表达，共聚焦显微镜观察发现两者存在共定位。综上表明，CEBPB能显著抑制PDCoV复制，为基于CEBPB研制潜在的抗病毒药物提供了新的思路。

关键词: CEBPB, PDCoV, Nsp4, 抗病毒

Abstract:

This study aims to explore the impact of CCAAT/enhancer-binding protein beta (CEBPB) on porcine deltacoronavirus (PDCoV) replication. PDCoV was inoculated in LLC-PK1 cells, and the expression levels of CEBPB at different time points were detected. Overexpression and knockdown of CEBPB were performed, and their effects on PDCoV replication were assessed using qPCR and Western blot. Furthermore, the role of CEBPB in the PDCoV replication cycle was clarified. Viral proteins that influenced the upregulation of CEBPB were screened using qPCR, and the colocalization of PDCoV viral proteins with CEBPB was observed using confocal microscopy. The results showed that CEBPB expression increased with prolonged virus infection time. Overexpression of CEBPB significantly inhibited PDCoV replication, while knockdown of CEBPB promoted virus replication. CEBPB mainly acted during the replication stage of PDCoV, and PDCoV Nsp4 could promote CEBPB expression. Confocal microscopy revealed colocalization of Nsp4 and CEBPB. In conclusion, CEBPB could significantly inhibit PDCoV replication in vitro, which provided important data for the development of antiviral drugs based on CEBPB.

Key words: CEBPB, PDCoV, Nsp4, anti-virus

中图分类号:

S852.659.6

陈琢琦, 范丽原, 钟纯燕, 于岩飞, 李基棕, 李彬, 袁晓民. 转录因子CEBPB对猪δ冠状病毒体外复制的抑制作用[J]. 畜牧兽医学报, 2025, 56(7): 3423-3432.

CHEN Zhuoqi, FAN Liyuan, ZHONG Chunyan, YU Yanfei, LI Jizong, LI Bin, YUAN Xiaomin. The Inhibitory Effect of Transcription Factor CEBPB on the Replication of Porcine Deltacoronavirus in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(7): 3423-3432.

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基因名称 Gene name	引物名称 Primer name	引物序列(5′→3′) Primer sequences (5′→3′)
PDCoV N	PDCoV-N-F	ATCGACCACATGGCTCCAA
	PDCoV-N-R	CAGCTCTTGCCCATGTAGCTT
	PDCoV-probe	CACACCAGTCGTTAAGCATGGCAAGCT
GAPDH	GAPDH-F	TCACTGCCACCCAGAAGACT
	GAPDH-R	ATGACCTTGCCCACAGCCTT
CEBPB	CEBPB-F	CTGGAGACGCAGCATAAGGT
	CEBPB-R	TGCTTGAACAAGTTCCGCAG
siCEBPB	siRNA-sense	CGCCUGCCUUUAAAUCCAUTT
	siRNA-antisense	AUGGAUUUAAAGGCAGGCGTT
si-NC	siRNA-sense	UUCUCCGAACGUGUCACGUTT
	siRNA-antisense	ACGUGACACGUUCGGAGAATT

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转录因子CEBPB对猪δ冠状病毒体外复制的抑制作用

The Inhibitory Effect of Transcription Factor CEBPB on the Replication of Porcine Deltacoronavirus in vitro

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