The effect and mechanism of sanguinarine against PCV2 based on the analysis of network pharmacology and TMT quantitative proteomics

Porcine circovirus type 2 (PCV2) is highly prevalent in nature and serves as the primary pathogen responsible for porcine circovirus-associated disease (PCVD/PCVAD), posing a significant threat to pig production. Currently, vaccination alone could not provide the complete protection for PCV2 infection. The active ingredients of traditional Chinese medicine have shown a positive effect in combating viral infections. This study employed tandem mass tag (TMT) labeled proteomic analyses and network pharmacology to examine the effect and mechanism of sanguinarine against PCV2. IFIH1, IFITM1, p38 alpha, and JNK were identified as the key targets of sanguinarine against PCV2 based on proteomics and network pharmacology. Using PCV2-infected PK-15 cells, it was discovered that sanguinarine inhibited the expression of the PCV2 CAP gene by upregulating IFIH1, thereby promoting STAT1 phosphorylation and activating MAVS expression. This, in turn, facilitated IRF3 phosphorylation, leading to increase IFITM1 expression. Simultaneously, sanguinarine suppressed the expression of the PCV2 CAP gene by inhibiting the expression of p38 alpha, JNK, and p-JNK protein. In conclusion, the results of this study suggest that sanguinarine exerts anti-PCV2 effects through different targets and pathways, which lays the foundation for the subsequent development of new anti-PCV2 veterinary drugs.