Chrysin inhibits PEDV replication by antagonizing apoptosis via the ROS/JNK/p53 axis.

Porcine epidemic diarrhea virus (PEDV) remains a significant threat to global swine industry, with immunization strategies limited by viral genome variation and a lack of effective therapeutic drugs. The flavonoid compound chrysin has shown potential anti-PEDV activity, but its specific antiviral mechanisms remain unclear. This study validated the inhibitory effect of chrysin on PEDV using IFA and qPCR. Additionally, Transcriptome sequencing was conducted on PEDV-infected Vero cells with and without chrysin treatment to reveal its antiviral mechanism. The results showed that 5813 DEGs at 12 hpi and 3374 DEGs at 24 hpi, with enrichment in pathways related to transcription, translation, metabolism, innate immunity, and apoptosis. Notably, KEGG analysis indicated that Chrysin regulate PEDV-induced apoptosis by modulating the TNF, MAPK, and p53 signaling pathways, which might antagonize PEDV infection. Subsequent results confirmed that chrysin can significantly downregulate the reactive ROS levels and cell apoptosis rate induced by PEDV infection (P<0.05), and inhibit the activation of the JNK and p53 pathways, including the phosphorylation of ASK1, JNK, and p53. Chrysin also significantly reduced cleaved caspase 3/8/9 levels and Bax/Bcl-2 ratios, thereby affecting PEDV N protein expression (P < 0.05). Moreover, similar inhibitory effects on PEDV can also be achieved through the JNK inhibitor SP600125 and the p53 inhibitor PFT-α. These findings revealed that chrysin can antagonize PEDV-induced apoptosis through the ROS/JNK/p53 signaling axis, thereby inhibiting viral replication. This study provides a scientific basis for developing chrysin as a novel anti-PEDV drug and offers insights into the screening and research of antiviral drug targets.