白藜芦醇对轮状病毒感染猪肠上皮细胞IPEC-J2的抑制效应

doi:10.11843/j.issn.0366-6964.2024.09.045

Abstract

Abstract:

The aim of this study was to investigate the antiviral effect of resveratrol on porcine rotavirus (PoRV) infected porcine intestinal epithelial cells(IPEC-J2). In this experiment, IPEC-J2 cells were used as research object. Firstly, the effect of different concentration of resveratrol on the cell viability of IPEC-J2 was determined. Then experimental groups including negative control group, PoRV-infected group and PoRV-infected group with the addition of resveratrol at different time points were set up depending on whether the IPEC-J2 cells were treated with PoRV or resveratrol (100 μmol·L^-1). The replication and proliferation of PoRV in IPEC-J2 cells were detected by real-time fluorescence quantitative PCR, viral titer assay and Western blot. The results showed that: 1) The addition of resveratrol (100 μmol·L^-1) significantly inhibited PoRV replication and infection at the late and full phases of viral infection of PoRV infection with IPEC-J2 (P < 0.05); 2) Compared with the PoRV-infected group, the addition of resveratrol (100 μmol·L^-1) during the full phase of PoRV-infected IPEC-J2 extremely significantly suppressed the levels of inflammatory cytokines IL-1β, IL-10, TNF-α and IFN-β in the supernatant fluid of IPEC-J2 cells (P < 0.01), while significantly suppressed the IL-10 and IFN-β levels during the late phase (P < 0.05); Extremely significantly suppressed the relative expression of immune-related factors MDA5, RIG-I, TRIF, MAVS, LGALS9, EIF2AK2, IRF9 and IFI44L in IPEC-J2 cells (P < 0.01) during late and full phases; Extremely significantly suppressed the relative mRNA expression of variable splicing factors SRPK1, SRPK2, HNRNPC and HNRNPR in IPEC-J2 cells (P < 0.01) during late and full phases; 3) Compared with the negative control group, PoRV infection significantly promoted exon jumps in exon 5 of LGALS9 and exon 2 of EF2AK2 in IPEC-J2 cells, while the addition of resveratrol (100 μmol·L^-1) significantly inhibited the exon skipping of target genes in the late and full phases of PoRV infection IPEC-J2 cells. In conclusion, this study confirmed that the addition of 100 μmol·L^-1 resveratrol could inhibit the infection and replication of PoRV in IPEC-J2 cells and play a role in the late phases of PoRV replication, which can provide a new basis and reference for the prevention and treatment of viral diarrhea in piglets.

Key words: resveratrol, porcine rotavirus, antiviral, alternative splicing

CLC Number:

S828.9

Ning PENG, Yaxu LIANG, Fei LONG, Dongming YU, Xiang ZHONG. Inhibitory Effect of Resveratrol on Rotavirus-infected Porcine Intestinal Epithelial Cells IPEC-J2[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 4213-4225.

Figures/Tables 8

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References 35

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基因 Gene name		引物序列(5′→3′) Primer sequence (5′→3′)	登录号 GenBank number
NSP5	Forward	CTGCTTCAAACGATCCACTCAC	MG451386
	Reverse	TGAATCCATAGACACGCC
MDA5	Forward	GCCACAGATCAGCCAAGTCT	MF358967
	Reverse	AGCTGACACTTCCTTCTGCC
RIG-I	Forward	GAATTTCCGGTCTGCCCTGA	NM_213770
	Reverse	GTCTGGCTCAGGAAAGTCCC
TRIF	Forward	TGACGTCACAGACCAGAAGC	MK302497
	Reverse	GGGGAGCTGAAGAAAGGGTC
MAVS	Forward	CTACTGCCTCCTCCTCCACT	MK302496
	Reverse	CAAATGCTGAGTTGGTGGGC
IRF9	Forward	GTTACCACTGAGGCCCCTTT	NM_001078670
	Reverse	GCAGCAGCGAGTAGTCTGAAT
IFI44L	Forward	GGCCCTATGCAGACTTGGTT	XM_021096421
	Reverse	CTATGGCCTGGCGAGTCAAA C
SRPK1	Forward	CATGGAGCGGAAAGTGCTTG	XM_021098048
	Reverse	CCTCGGTGCTGAGTTTCAGA
SRPK2	Forward	ATGAGCTCCCGGAAAGTGC	XM_021102583
	Reverse	TGTTGAGGCTCCGGCTTTTT
HNRNPC	Forward	GTCCCCTCTACTCAGCTCCT	XM_021098264
	Reverse	AACACGTGCTGGGTAACTGT
HNRNPR	Forward	GCCCTGGTAAACACCTTGGA	XM_021098239
	Reverse	ACCCTCTGTTAAACCTGTGACT
GAPDH	Forward	GGAGAACGGGAAGCTTGTCA	NM_001206359
	Reverse	GCCTTCTCCATGGTCGTGAA
LGALS9	Forward	GGACTGGACCCTGCACTTC	NM_214319
	Reverse	AGAAACACGGTGAACGACCA
EIF2AK2	Forward	GACAACCAGGAGCCACAAGG	XM_02108095
	Reverse	AAACACGGTGAACGACCACT

抗体 Antibodies	编号 Identifier	来源 Source	宿主 Host	稀释比例 Dilution rate
VP6	定制	Abconal	兔	1∶5 000
GAPDH	60004-1-Ig	Proteintech	兔	1∶10 000
山羊抗兔IgG	ab205718	Abcam	兔	1∶10 000

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Inhibitory Effect of Resveratrol on Rotavirus-infected Porcine Intestinal Epithelial Cells IPEC-J2

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