猪伪狂犬病病毒实验感染江口萝卜仔猪的脑组织转录组学差异分析

doi:10.11843/j.issn.0366-6964.2025.10.036

摘要/Abstract

摘要：

为了研究猪伪狂犬病病毒(PRV)感染对江口萝卜猪脑组织基因表达的影响。将12头2月龄仔猪随机分为感染组和对照组，感染组通过肌肉注射接种1 mL PRV-HLJ株(毒价为1×10⁴ TCID₅₀ ·100 μL^-1)，对照组注射等量PBS。感染后7 d，采集脑组织进行组织病理学、免疫组织化学和转录组学测序分析。通过基因本体论(GO)和京都基因与基因组百科全书(KEGG)分析差异表达基因的功能和通路，并使用荧光定量PCR(qRT-PCR)验证关键基因的表达趋势。结果显示：感染组脑组织中少突胶质细胞数量显著增加，且PRV阳性表达明显。转录组测序共鉴定出269个差异表达基因，其中149个基因上调，120个基因下调。GO分析显示，差异基因主要涉及信号转导、跨膜转运、凋亡和神经活性配体-受体相互作用等生物过程。KEGG分析表明，差异基因显著富集于神经活性配体-受体相互作用、铁死亡和IL-17信号通路。qRT-PCR结果显示，神经活性配体-受体相互作用通路中的LPAR3、GZMA、P2RY2和NMB基因下调表达，而CNR1、CALCR和NTSR1基因上调表达；铁死亡通路中的SLC7A11和SLC40A1基因上调表达，TF基因下调表达；IL-17信号通路中的FOS、FOSB和MAPK15基因均下调表达。本试验研究发现为进一步研究PRV的致病机制提供了新的分子生物学证据，并为PRV的防控和治疗策略提供了重要数据参考。

关键词: 猪伪狂犬病病毒, 江口萝卜猪, 脑组织, 转录组学, 致病机理

Abstract:

To investigate the impact of porcine pseudorabies virus (PRV) infection on gene expression in the brain tissue of Jiangkou radish pigs, 12 two-month-old piglets were randomly divided into an infected group and a control group. The infected group was intramuscularly inoculated with 1 mL of the PRV-HLJ strain (with a titer of 1×10⁴ TCID₅₀ ·100 μL^-1), while the control group received an equal volume of PBS. Seven days post-infection, brain tissues were collected for histopathological, immunohistochemical, and transcriptomic sequencing analyses. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted to explore the functions and pathways of differentially expressed genes, and quantitative real-time PCR (qRT-PCR) was used to validate the expression trends of key genes. The results showed a significant increase in the number of oligodendrocytes in the brain tissue of the infected group, along with clear PRV-positive expression. Transcriptomic sequencing identified a total of 269 differentially expressed genes, including 149 upregulated genes and 120 downregulated genes. GO analysis revealed that these genes were primarily involved in biological processes such as signal transduction, transmembrane transport, apoptosis, and neuroactive ligand-receptor interactions. KEGG analysis indicated that the differentially expressed genes were significantly enriched in pathways related to neuroactive ligand-receptor interactions, ferroptosis, and the IL-17 signaling pathway. qRT-PCR results demonstrated that in the neuroactive ligand-receptor interaction pathway, the genes LPAR3, GZMA, P2RY2, and NMB were downregulated, while CNR1, CALCR, and NTSR1 were upregulated. In the ferroptosis pathway, SLC7A11 and SLC40A1 were upregulated, while TF was downregulated. In the IL-17 signaling pathway, FOS, FOSB, and MAPK15 were all downregulated. These findings of this study provide new molecular evidence for further research into the pathogenic mechanisms of PRV and offer important data references for the prevention, control, and treatment strategies of PRV.

Key words: pseudorabies virus, Jiangkou radish piglets, brain tissue, transcriptomics, pathogenic mechanism

中图分类号:

S858.285.3

孙伟, 刘杉杉, 樊雨, 邓生青, 王清艳. 猪伪狂犬病病毒实验感染江口萝卜仔猪的脑组织转录组学差异分析[J]. 畜牧兽医学报, 2025, 56(10): 5170-5179.

SUN Wei, LIU Shanshan, FAN Yu, DENG Shengqing, WANG Qingyan. Transcriptomic Differential Analysis of Brain Tissues in Jiangkou Radish Piglets Experimentally Infected with Pseudorabies Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(10): 5170-5179.

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基因名称 Gene name	GenBank登录号 GenBank accession number	引物名称 Primer name	引物序列(5→3′) Sequence	通路名称 Pathway name
LPAR3	NM_001162402.1	LPAR3-F LPAR3-R	CATGTCAGTCACGAGTATGCGGATC AGAGCAGGAAGAGATGTCACAGAGG	Neuroactive ligand-receptor interaction
GZMA	NM_001143709.1	GZMA-F GZMA-R	TGCTTTGACCAGGACACACATGAG CCACTTTAGGGAGATGAAGGACAGC
P2RY2	NM_001006591.1	P2RY2-F P2RY2-R	CGCTCTACTTCATCACCACCACTG AGACCAGGATGACGGCGAAGG
NMB	XM_021097907.1	NMB-F NMB-R	GGACAACGCACCACATCTCTCTG ACCAGCAGCCTCCTGTACGG
CNR1	XM_013992675.2	CNR1-F CNR1-R	GGTACTGTGCGTTATCCTCCATTCC CGACGAAGCTGTAGACGAAGATGAC
NTSR1	XM_021078238.1	NTSR1-F NTSR1-R	GTGGTGTCCGTGTTCGCTGAG TGGCATGAGCAGTAGTTGGAATCAG
CALCR	NM_214354.2	CALCR-F CALCR-R	GTGGTGTCCGTGTTCGCTGAG TGGCATGAGCAGTAGTTGGAATCAG
SLC7A11	XM_021101587.1	SLC7A11-F SLC7A11-R	TCTTTGTTGCCCTCTCCTGCTTTG GTGTGTTTGCGGATGTGAATCATGG	Ferroptosis
TF	NM_001244653.1	TF-F TF-R	TCTAAGATGGACTCCTCGCTGTACC GGCACACCACCTCACCTTCTTG
SLC40A1	XM_013984335.2	SLC40A1-F SLC40A1-R	TGTCCCTGAGATGAGCCCTAAACC AAGACCGATTCTAGCAGCAATGACG
FOS	NM_001123113.1	FOS-F FOS-R	CTGAGATCGCCAACCTGCTGAAG CAGATCAAGGGAAGCCACAGACATC	IL-17 signaling pathway
FOSB	XM_063252899.1	FOSB-F FOSB-R	ACCCCTACGACATGCCAGGAAC ACCACCGCCACTAGCTCCAC
MAPK15	XM_021090469.1	MAPK15-F MAPK15-R	TGAGTGGACACTAGAGGCGGATG TGATAGAGGCGGCGGCGATAC
β-actin	ON164673.1	β-actin-F β-actin-R	TCGCCGACAGGATGCAGAAG CAGGATGGAGCCGCCGATC

通路名称 Pathway Name	基因名称 Gene Name	Fc	log₂(Fc)	调节 Regulation
神经活性配体-受体相互作用 Neuroactive ligand-receptor interaction	LPAR3	-3.57	0.08	down
	GZMA	-5.25	0.03	down
	P2RY2	-1.36	0.39	down
	NMB	-1.70	0.31	down
	CNR1	1.14	2.21	up
	CALCR	1.33	2.51	up
	NTSR1	1.61	3.05	up
铁死亡 Ferroptosis	SLC7A11	3.74	1.90	up
	TF	0.46	-1.12	down
	SLC40A1	2.13	1.09	up
IL-17信号通路 IL-17 signaling pathway	FOS	0.31	-1.69	down
	FOSB	0.36	-1.48	down
	MAPK15	0.44	-1.18	down

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