Exploration of the effect and mechanism of indirubin on infectious bovine rhinotracheitis virus based on network pharmacology

Our team’s previous research indicates that infectious bovine rhinotracheitis virus (IBRV) induces mitochondrial damage in bovine kidney cells (MDBK). Indirubin, a bisindole alkaloid, can alleviate mitochondrial damage. However, it is currently unclear whether indirubin can regulate mitochondrial damage caused by IBRV in MDBK cells. This study predicted the anti-IBRV effect of indirubin through network pharmacology, verified its anti-IBRV activity through experiments, and confirmed that indirubin can alleviate mitochondrial damage caused by IBRV. Network pharmacological results show that 81 potential targets of indirubin in infectious bovine rhinotracheitis (IBR) have been screened through network analysis. The analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) shows that the anti-IBR activity of indirubin involves multiple signaling pathways. Genetic ontology (GO) analysis reveals that its anti-IBR effect encompasses a variety of biological functions, with 15 target genes enriched in mitochondria. Virus suppression experiments have shown that indirubin can inhibit IBRV virus replication in MDBK cells. Flow cytometry showed that indirubin can reduce the elevated levels of reactive oxygen species (ROS) and depolarization of mitochondrial membrane potential (MMP) caused by IBRV infection. The molecular docking results confirm that indirubin exhibits a strong binding affinity with 15 targets. Our research shows that indirubin may exert anti-IBRV properties by regulating the production of ROS and inhibiting MMP depolarization.