PROTECTIVE EFFECTS OF A RODGERSIA SAMBUCIFOLIA FLAVONOID–SCUTELLARIA POLYSACCHARIDE COMPOUND AGAINST PSEUDORABIES VIRUS-INDUCED TESTICULAR INJURY VIA MODULATION OF THE FOXO SIGNALING PATHWAY

Pseudorabies virus (PRV) is known to induce severe reproductive disorders, yet the molecular mechanisms underlying testicular damage remain unclear. This research aims to explore the impact of PRV infection on porcine testicular (ST) cells and murine testicular tissue, with a focus on the involvement of the FOXO signaling pathway. This study also evaluated the antagonistic effect of compound RS on PRV infection through regulation of this pathway. Analyze differentially expressed genes and enriched pathways in ST cells infected with PRV through transcriptome sequencing. ST cells were infected with PRV and subsequently treated with either the FOXO pathway inhibitor AS1842856 or varying concentrations of RS (12.5, 25, and 50 μg/mL) to evaluate their effects on viral replication and FOXO gene expression. Cell viability was detected by CCK-8, viral load and mRNA expression of FOXO were detected by qPCR. Establish a PRV infected mouse model, observe pathological changes in testicular tissue through HE staining, detect testicular virus concentration, sperm count, and FOXO gene expression. PRV infection caused significant cytopathic effects in ST cells, with transcriptome sequencing identifying 1,650 significantly upregulated genes and 1,359 significantly downregulated genes (|log2FC| ≥ 1, P < 0.01). KEGG pathway analysis indicated significant enrichment of these genes in the FOXO signaling pathway, among others. Inhibition of the FOXO pathway markedly reduced viral replication in ST cells. PRV infection upregulated FOXO1 and FOXO4 mRNA expression, while downregulating FOXO3; these alterations were reversed upon RS treatment. In vivo experiments, RS treatment alleviated the pathological damage of mouse testicular tissue caused by PRV, reduced virus concentration in the testes, and partially restored sperm count. Compound RS effectively suppresses PRV replication both in vitro and in vivo by modulating the FOXO pathway and alleviates testicular injury caused by PRV. These findings provide a theoretical basis and identify a promising candidate for developing novel therapeutics targeting PRV-associated reproductive toxicity. Keywords: Rodgersia sambucifolia hemsl flavon; Scutellaria polysaccharide; Pseudorabies virus; FOXO signaling pathway