红景天苷对犬细小病毒体外复制的抑制效应分析

doi:10.11843/j.issn.0366-6964.2024.12.036

Abstract

Abstract:

This study analyzes the inhibitory effect and its mechanism of salidroside (SAL) on canine parvovirus (CPV). In vitro, drug effects were performed on three stages of virus infection (adsorption, invasion, and replication). Virus titers were detected in cells to access the inhibitory effect of SAL on the virus were evaluated. TUNEL (Terminal deoxynucleotidyl Transferase dUTP Nick End Labeling) assay was used to analyze apoptosis. Quantitative Real-time PCR and Western blot were used to detect the mRNA or protein expression of apoptosis-related proteins and inflammatory factors, respectively, aiming to analyze and preliminarily verify the mechanism by which SAL inhibits virus replication. The results showed that SAL significantly inhibited CPV replication, but had no significant effect on the adsorption and invasion processes of CPV. SAL could inhibit apoptosis induced by CPV and significantly inhibit the protein expression of caspase 8 and tBID. Further research revealed that CPV could induce the upregulation of IL-1β and inflammatory factors, while SAL downregulated the expression of some inflammatory factors in virus-infected cell. The activation of caspase 1 and NLRP3 was closely related to IL-1β. After viral infection of cells, incubation with SAL could inhibit the activation of caspase 1, but had no significant effect on NLRP3. When virus was incubated in cells with down-regulated expression of caspase 8 by siRNA-caspase 8, the expression of IL-1β in cells was inhibited, consistent with the effect of SAL, indicating that SAL mainly inhibits the expression of inflammatory factors by inhibiting the activation of the caspase 8 signaling pathway. In summary, this study shows that SAL can effectively inhibit the replication of CPV. SAL inhibits CPV-induced apoptosis by regulating the caspase 8 signaling pathway and inhibits the expression of some inflammatory factors. This study provides a new approach and method for the effective treatment of CPV.

Key words: canine parvovirus(CPV), salidroside, apoptosis, cytokines

CLC Number:

S852.659.2

ZHANG Meiwen, WANG Chenglong, LIU Yuzhen, ZHAO Yutong, ZHU Jiping, LI Yi. Analysis of the Inhibitory Effect of Salidroside on Canine Parvovirus Replication in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5738-5750.

Figures/Tables 13

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基因 Gene	正向引物(5′→3′) Forword primer(5′→3′)	反向引物(5′→3′) Reverse primer(5′→3′)
Caspase 8	ACAAGGGCATCATCTATGGCTCTGA	CCAGTGAAGTAAGAGGTCAGCTCAT
Caspase 12	GCCGTCTGGGTGACTGATG	CTGCAAGGGCTGGTCACAT
p53	TAACAGTTCCTGCATGGGCGGC	AGGACAGGC ACAAACACGCACC
Bax	TTCCGAGTGGCAGCTGAGATGTTT	TGCTGGCAAAGTAGAAGAGGGCAA
Bcl2	CATGCCAAGAGGGAAACACCAGAAG	GTGCTTTGCATTCTTGGATGAGGG
FasL	GAGGAGGGACCACAACACAGGTCTC	AGAGCTGAAACATCCCCAGC
Fas	ACTGTGCAGACATCGACCTG	CGTTGATCCCGTTCTTCCGA
FADD	TGGAGGAGACTGGCTCGTTA	CTTGTTGGCTGGCTCCAAAC
IL-8	TGGAAATGAGGTGTGCCTGG	GGATCTTGTTTCTCAGCCTTCTT
IL-7	CAGCATCAATGACTTGGACATCA	TTCAACTTGTGAGCAGCACG
IL-6	CTCCTGGTGATGGCTACTGC	GTGCAGAGATTTTGCCGAGG
TNF-α	GTGCCGTCAGATGGGTTGTA	CAAGGGCTCTTGATGGCAGA
CCL5	GACTACCCTTCACCCACGTC	CTTTCGGGTGACAAAGACGAC
CXCL10	TGACTCTGAATGGTACTCAAGGA	GACACGATGGACTTGCAGGA
IFNB	GCGAGAAATCACGCCAGTTC	TCTATTGTCCAGGCACAGATGC
GAPDH	GCTGCCCAGAACATCATCC	GTCAGATCCACGACGGACAC

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Analysis of the Inhibitory Effect of Salidroside on Canine Parvovirus Replication in vitro

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