Coptisine-naringenin combination inhibits PRRSV replication via the MET/PI3K/Akt pathway thereby attenuating virus-induced mitochondrial apoptosis

In our previous work, six natural compounds-ferulic acid, kaempferol, choline, coptisine, quercetin, and naringenin-were identified to inhibit PRRSV replication in Marc-145 and PAM cells. In this study, we performed a systematic combination optimization of these six compounds to develop a synergistic anti-PRRSV formulation.

First, network pharmacology was employed to predict the potential targets of the six compounds against PRRSV, and differential target profiles guided the initial design of compound combinations. Subsequently, a six-factor, three-level orthogonal design was applied to construct multi-compound formulations, and the Chou-Talalay method was used to evaluate two-compound combinations. The safety of each formulation was assessed in Marc-145 cells by CCK-8 assay, and the anti-PRRSV efficacy was determined by RT-qPCR and Western blot analysis of PRRSV N gene and protein expression.