A multiomics study uncovers the immune-enhancing effects and mechanisms of Zhenqi granules

Abstract Many nations have prohibited the use of antibiotic growth promoters because of concerns regarding the increased development of antimicrobial resistance and the risks associated with drug residues. Traditional Chinese medicines (TCMs) are valuable alternatives to antibiotics. Our previous research demonstrated that Zhenqi granules (ZQ), a formula containing Huangqi and Nvzhenzi, significantly promoted the growth of pigs. This study further investigated the fundamental mechanism from the perspective of immune enhancement by combining transcriptomics, network pharmacology, and cell experiments. Piglet experiments revealed that ZQ administration significantly increased the production of antibodies against CSFV following vaccination. Moreover, compared with control piglets, ZQ-treated piglets presented significantly increased serum cytokine levels and peripheral blood lymphocytes proliferation, indicating the immune-enhancing effect of ZQ. Comparative transcriptomic analysis of spleen tissues revealed that ZQ administration altered the expression of numerous immunity-related genes, with significant enrichment in the calcium signaling pathway. Network pharmacology analysis demonstrated that multiple active components of ZQ interact with immune-related targets, particularly those in the calcium signaling pathway. In vitro cell experiments confirmed that ZQ treatment activated key proteins and downstream transcription factors in this pathway. These findings provide mechanistic insights into the immune-enhancing effect of ZQ, in which the calcium signaling pathway plays a key role.