Abstract
BackgroundThis study aimed to explore the underlying mechanism of puerarin against rotavirus (RV), based on network pharmacology analysis and experimental study in vitro.MethodsThe cytopathic effect inhibition assay with different concentrations of puerarin at different times of infection in vitro was applied to evaluate the effect of puerarin against human RV G1P[8] Wa strain (HRV Wa). Subsequently, the potential targets of puerarin and RV-related genes were obtained from online databases. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the major target genes were also performed. Furthermore, the major targets and signaling pathway related to RV infection were verified at the molecular level via Western blot, quantitative real-time reverse transcription PCR (RT-qPCR), and Enzyme-linked immunosorbent assay (ELISA) tests.ResultsOur results suggest that puerarin had a certain inhibitory effect on viral replication and proliferation. The network pharmacology analysis showed that a total of 436 puerarin corresponding target and 497 RV-related targets were acquired from the online databases. The core targets of puerarin against RV, such as Toll-like receptor 4 (TLR4), tumor necrosis factor (TNF), and caspase 3 (CASP3), were identified from the protein-protein interaction (PPI) network. The KEGG analysis indicated that the TLR signaling pathway was one of the crucial mechanisms of puerarin against RV. In particular, puerarin could inhibit the expression of key factors of the TLR4/nuclear factor kappa-B (NF-κB) signaling pathway in HRV-infected Caco-2 cells and regulate the levels of cellular inflammatory factors.ConclusionsBased on the network pharmacology analysis and experimental study, the study showed that puerarin not only had an anti-RV effect, but could also modulate the inflammatory response induced by RV infection via the TLR4/NF-κB signaling pathway. This study reveals the potential of puerarin in the treatment of RV infection, suggesting that it might be a promising therapeutic agent.
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