Dihydroartemisinin modulates the host proteome to suppress bovine viral diarrhea virus via mitochondrial and ER stress.

Bovine Viral Diarrhea Virus (BVDV) remains a pervasive threat to cattle health and food security, yet the host pathways it exploits are only partially understood. Here, we employed high-resolution Orbitrap Astral FAIMS-DIA proteomics to interrogate Madin-Darby bovine kidney cells in three states: uninfected control (CON), BVDV-infected (CP), and BVDV-infected cells treated with dihydroartemisinin (DHA; CP_D). Quantification of more than 7,000 proteins revealed a conserved 98% core proteome, but sharply divergent condition-specific signatures. Infection alone upregulated complement coagulation and cytokine-receptor pathways, highlighting innate immune activation alongside zinc finger-rich transcriptional remodeling. DHA superimposed a distinct proteomic landscape, roughly doubling the number of differentially abundant proteins in the cytoplasm, mitochondria, and extracellular space, and markedly increasing the number of nuclear proteins. Enrichment analyses revealed a significant shift toward extracellular matrix receptor interactions, hematopoietic cell lineages, and kinase-dominated signaling. At the same time, network centrality identified newly emergent hubs within the protein kinase and immunoglobulin superfamilies. Domain-level evidence, combined with expanded metabolic pathway coverage, suggests that DHA orchestrates the parallel rewiring of signal transduction, matrix architecture, and bioenergetics, rather than merely reversing viral perturbations. Principal component clustering and hierarchical correlations confirmed that CP_D forms a reproducible proteomic state distinct from both CON and CP, arguing for a multimodal host reprogramming mechanism. Collectively, these data position DHA as a host-directed antiviral that partitions the proteome into defensive, signaling, and structural modules, offering a blueprint for multi-target intervention against pestiviral disease.