紧密连接蛋白CLDN4促进猪流行性腹泻病毒感染

doi:10.11843/j.issn.0366-6964.2025.06.026

Abstract

Abstract:

Tight junction proteins play a crucial role in various viral infections, with tight junction protein 4 (Claudin-4, CLDN4) being a major member of this family. The role of CLDN4 in porcine epidemic diarrhea virus (PEDV) infection remains unclear. To explore the impact of PEDV infection on CLDN4 expression and the role of CLDN4 in PEDV infection, we performed quantitative PCR (qPCR) and Western blot to assess the transcriptional and protein expression levels of CLDN4 following PEDV infection. Additionally, a eukaryotic expression plasmid for the CLDN4 gene was constructed, and specific interference RNA targeting CLDN4 was designed. The effects of overexpression and knockdown of CLDN4 on PEDV replication were analyzed using qPCR, Western blot, immunofluorescence assay (IFA), and PEDV viral titer measurements. Finally, we examined the effects of overexpression of CLDN4 on the adhesion and invasion phases of PEDV infection. Results showed that PEDV infection upregulates CLDN4 transcription and protein levels, while overexpression of CLDN4 significantly promotes PEDV replication, and interference with CLDN4 expression inhibits PEDV replication. Moreover, CLDN4 facilitates PEDV adhesion and invasion. This study suggests that CLDN4 plays a role in promoting PEDV infection and replication, providing a reference for investigating CLDN4 function and elucidating PEDV replication mechanisms.

Key words: CLDN4, porcine epidemic diarrhea virus, promote, infection

CLC Number:

S852.659.6

LI Chengcheng, ZHAO Yongxiang, CAO Qiuxia, SONG Xu, LI Yupeng, FAN Baochao, GUO Rongli, XU Yefen, LI Bin. Tight Junction Protein CLDN4 Promotes Porcine Epidemic Diarrhea Virus Infection[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(6): 2826-2835.

Figures/Tables 6

Table 1

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References 31

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引物/siRNA名称 Primer/siRNA names	序列(5'→ 3') Sequence	用途 Aapplication
CLDN4-F	CTAG$\underline{{\rm{TCTA}}}$GAGCCACCATGGCCTCCATGGGGCTAC	扩增CLDN4
CLDN4-R	CCG$\underline{{\rm{CTCGAG}}}$TCAGATTACAAGGACGACGATGACAAGCACGTAGTTGCTGGCAGC	基因
Q-CLDN4-F	CAGTGCAAGGTGTACGACTC	CLDN4
Q-CLDN4-R	TCATCCTCCAGGCAGTTTGT	基因定量
Q-PEDV-N-F	CGCAAAGACTGAACCCACTAACTT	PEDV-N
Q-PEDV-N-R	TTGCCTCTGTTGTTACTCGGGGAT	基因定量
Q-GAPDH-F	TCACTGCCACCCAGAAGACT	GAPDH
Q-GAPDH-R	ATGACCTTGCCCACACAGCCTT	基因定量
si-CLDN4-F1	CCCUCGUCAUCAUCAGCAUTT	siRNA干扰
si-CLDN4-R1	AUGCUGAUGAUGACGAGGGTT	序列1
si-CLDN4-F2	CGCACAGACAAGCCUUACUTT	siRNA干扰
si-CLDN4-R2	AGUAAGGCUUGUCUGUGCGTT	序列2
si-NC-F	UUCUCC GAACGUGUCACGUTT	干扰对照
si-NC-R	ACGUGACACGUUCGGAGAATT	序列

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Tight Junction Protein CLDN4 Promotes Porcine Epidemic Diarrhea Virus Infection

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