猪流行性腹泻病毒感染仔猪空肠转录组差异表达分析

doi:10.11843/j.issn.0366-6964.2024.06.036

摘要/Abstract

摘要：

旨在分析猪流行性腹泻病毒(PEDV)感染仔猪空肠组织的转录组差异。用PEDV毒株HB(G2b)感染3头2日龄仔猪，对感染后18 h仔猪空肠组织进行免疫组化、转录组测序和分析，最后采用荧光定量PCR(RT-qPCR)验证转录组结果。结果显示：仔猪感染PEDV 18 h后空肠绒毛明显脱落、变短，且PEDV主要分布于空肠肠绒毛表面；GO富集及KEGG富集分析的差异基因主要与免疫应答过程和炎症反应过程相关；富集在趋化因子受体相互作用、Toll受体信号、脂肪与糖代谢等。结果提示PEDV感染可激活宿主免疫反应及相关抗病毒信号通路，并且引起宿主的炎症风暴和营养代谢紊乱，为深入研究PEDV的致病机制奠定了基础。

关键词: 猪流行性腹泻病毒, 转录组, 致病机制

Abstract:

The aim is to study the pathogenic mechanism of porcine epidemic diarrhea virus (PEDV) by transcriptome analysis of the jejunum of two-day-old piglets infected with PEDV. Three two-day-old piglets were infected with the PEDV HB strain (PEDV-HB), which belongs to the currently prevalent G2b subtype. At 18 h post-infection, jejunum tissues were collected from piglets for pathology, PEDV detection, RNA-seq and transcriptome analysis. Finally; the transcriptome results were verified by quantitative real-time PCR (RT-qPCR). The result showed that the jejunal villi of piglets infected PEDV-HB were significantly shed and shortened; and was mainly distributed on the surface of intestinal villi. GO and KEGG enrichment analysis showed that differentially expressed genes (DEGs) were mainly related to immune response and inflammatory response, and were enriched in several classical pathways, such as cytokine-cytokine receptor interaction, Toll-like receptor signaling pathway, fat and glucose absorption and metabolism. PEDV infection of the jejunum of piglets activates the host immune response and associated antiviral signaling pathways and induces a cytokine storm as well as dysfunctions in fat and glucose absorption and metabolism. In all, this research can lay the foundation for the study of pathogenesis mechanism of PED.

Key words: porcine epidemic diarrhea virus, transcriptions, pathogenesis mechanisms

中图分类号:

S852.6596

李栋梁, 郑关民, 李帅, 朱洪森, 吴超. 猪流行性腹泻病毒感染仔猪空肠转录组差异表达分析[J]. 畜牧兽医学报, 2024, 55(6): 2652-2661.

Dongliang LI, Guanmin ZHENG, Shuai LI, Hongsen ZHU, Chao WU. Differential Expression of Transcriptome in Jejunal of Piglets Infected with Porcine Epidemic Diarrhea Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(6): 2652-2661.

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基因名称 Gene name	上游引物(5′→3′) Forward primer	下游引物(5′→3′) Reverse primer
IL-8	ACTGGCTGTTGCCTTCTTGG	CTGGCATCGAAGTTCTGCAC
CIAP	GTTCAGTTAACATTTCCGGAGCC	GGTCAATGGCCACATCTGGAA
NFKBIA	TCACCTACCAGGGCTACTCC	CCTCTGTGAACTCTGACTCCG
SYK	GGACAGAAGCCATATCGGGG	AGCTCCACCGCTACAAATCC
P13CABD	ACTGTGAACGAGCCTATGCC	CCGTTTATGGCTGTTGGGTTC
IL-6	GGTGATGGCTACTGCCTTCC	TGCAGAGATTTTGCCGAGGA
RIGI	TTTCCACGAGGACGAAAGGG	GTGTCAGACGTTGCTCCAGA
BAFF	AAAGCCCCTCTGTCCGCATC	CTGGTGGTGCAAAGATCCCTT
IL-1β	AAGATAACACGCCCACCCTG	GAGTTTCCCAGGAAGACGGG
CCL2	GCGGCTGATGAGCTACAGAAG	CCGCGATGGTCTTGAAGATC
CCL20	GGTGCTGCTGCTCTACCTCT	GCTGTGTGAAGCCCATGATA
CXCL2	CCGTGCAAGGAATTCACCTC	TGCGGGGTTGAGACAAACTT
CCL25	GCCTACCACAGCCACATTAAG	GCTTCCCGCACACCATCTT
CCL5	ATCAGCCTCCCCATATGCCT	CCGCACCCATTTCTTCTCTG
CCR7	CAATTTCAACATCACCAGCGG	GAACTTGACGCCGATGAAGG
CCL19	GTTCGAGCCTTTCGCTACCT	GTGGTGCTTGCTCTTAGCAG
CCL3L1	CTTCCTCGCAAATTCGTAGC	GCATTCAGCTCCAGGTCAG
CCL22	TTACAGCTGCTCCCTGACAC	TGGATTGGAAGGGTAAGAGG
GADPH	TCATCATCTCTGCCCCTTCT	GTCATGAGTCCCTCCACGAT

基因 Gene	基因ID GenBank ID	转录基因log₂FC RNA-seq log₂FC	荧光定量log₂FC RT-qPCR log₂FC
IL-8	NM_213867.1	2.08	1.34
CIAP	XM_021062707.1	2.38	0.90
NFKBIA	NM_001005150.1	1.50	0.94
SKY	NM_001104952.1	1.10	0.68
P13CABD	XM_021095292.1	1.79	2.54
IL-6	NM_214399.1	4.76	4.23
RIGI	XM_021064058.1	1.47	1.57
BAFF	NM_001097498.1	1.61	1.46
IL-1β	XM_021085847.1	2.49	1.44

基因 Gene	基因ID GenBank ID	转录基因log₂FC RNA-seq log₂FC	荧光定量log₂FC RT-qPCR log₂FC
CCL2	NM_214214.1	2.55	1.79
CCL20	NM_001024589.1	2.21	1.17
CXCL2	NM_001001861.2	2.56	2.50
CCL25	XM_005661258.3	-3.51	-4.32
CCL5	NM_001129946.1	0.38	0.63
CCR7	NM_001001532.3	1.49	2.35
CCL19	NM_001170516.1	2.39	1.81
CCL3L1	NM_001009579.1	3.28	2.53
CCL22	NM_001256776.2	-1.02	-0.45

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