单核细胞趋化蛋白诱导蛋白3对猪流行性腹泻病毒复制的体外抑制作用

doi:10.11843/j.issn.0366-6964.2025.06.028

Abstract

Abstract:

The purpose of this study was to investigate the effect of monocyte chemotactic protein-induced protein 3 (MCPIP3) on porcine epidemic diarrhea virus (PEDV) replication. The dynamic change of MCPIP3 transcription level after PEDV infection was determined by RT-qPCR. Then, using monkey kidney epithelial cell cDNA as template, ZC3H12C gene was amplified by PCR and cloned into P3×Flag-CMV-14 eukaryotic expression vector to construct MCPIP3 eukaryotic recombinant vector. MCPIP3 protein was overexpressed or interfered on African green monkey embryonic kidney cells (MARC-145), and the influence of MCPIP3 protein on PEDV replication was investigated by Western blot, quantitative PCR, and virus titer determination. The results showed that PEDV infection significantly up-regulated ZC3H12C transcription on MARC-145 cells. The MCPIP3 recombinant plasmid was successfully constructed, which could induce exogenous high expression of MCPIP3 protein after transfection, and the specific siRNA could significantly down-regulate the transcription level of ZC3H12C. Compared with the control group, high expression of MCPIP3 significantly inhibited PEDV N protein expression level, down-regulated PEDV N gene transcription levels and virus titer, while interfered with MCPIP3 expression had the opposite effect, indicating that MCPIP3 significantly inhibited PEDV proliferation on MARC-145 cells. Furthermore, the MCPIP3 expression negatively regulated PEDV induced transcription of IL-8, IL-12β and TNF-α. This study expanded the function of MCPIP3 protein, and provided a theoretical basis for the study of PEDV replication mechanism.

Key words: MCPIP3, PEDV, viral replication, antiviral effect

CLC Number:

S855.3

ZHOU Hongting, SUN Xinru, FAN Baochao, ZHANG Xuehan, ZHOU Jinzhu, SUOLANG Sizhu, SUN Min, GONG Ga, LI Bin. Inhibitory Effect of Monocyte Chemotactic Protein Induction Protein 3 on Porcine Epidemic Diarrhea Virus Replication in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(6): 2847-2856.

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基因 Gene	引物 Primer	序列(5'→3') Sequence
ZC3H12C	ZC3H12C-F	CGCCACCATGCCGGGTAGCGGCTCCCAGG
	ZC3H12C-R	ATAACCCAGCTGGGATTTCTCC
ZC3H12C-CHZ	ZC3H12C-CHZ-F	TGAACCGTCAGAATTAAGCTT CGCCACCATGCCGGGTAGCGGCTCCCAGG
	ZC3H12C-CHZ-R	TTTGTAGTCAGCCCGGGATCC ATAACCCAGCTGGGATTTCTCC

基因 Gene	干扰 RNA Interfering RNA	序列(5'→3') Sequence
ZC3H12C	si-MCPIP3-sense	CGUGCCAUGUCUAGAAAUATT
	si-MCPIP3-antisense	UAUUUCUAGACAUGGCACGTT
NC(Negative control)	NC-sense	UUCUCCGAACGUGUCACGUTT
	NC-antisense	ACGUGACACGUUCGGAGAATT

基因 Gene	引物 Primer	序列(5'→3') Sequence
m-GADPH	qm-GADPH-F	TGACCTGCCGTCTGGAAAA
	qm-GADPH-R	GGAAGAGTGGGTGTCGCTGT
m-ZC3H12C	qm-ZC3H12C-F	GAGAGTGGTGTGCTATGATGAC
	qm-ZC3H12C-R	CTCGTTAGCCAAATCCCTGTAG
m-IL-8	qm-IL-8-F	GACCACACTGCGTCAATACA
	qm-IL-8-R	AAACTTCTCCACAACCCTAGAC
m-IL-12β	qm-IL-12β-F	ACCAGAGTGGTGAGGTCTTA
	qm-IL-12β-R	CCTCCTTTGTGACAGGTGTATT
m-TNF-α	qm-TNF-α-F	AGCCCATGTTGTAGCAAACC
	qm-TNF-α-R	TGAGGTACAGGCCCTCTGAT
PEDV-N	PEDV-N-F	CGCAAAGACTGAACCCACTAACTT
	PEDV-N-R	TTGCCTCTGTTGTTACTCGGGGAT
	PEDV-N-probe	CTGTTGTTGCCATTGCCACGA

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Inhibitory Effect of Monocyte Chemotactic Protein Induction Protein 3 on Porcine Epidemic Diarrhea Virus Replication in vitro

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