Anti-inflammatory effects of 3PO in asthmatic airway inflammation: an integrated study using network pharmacology, molecular modelling, and in vivo experiments

Introduction: Recent studies have highlighted the critical role of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3 (PFKFB3) in inflammation, with inhibitors like 3PO showing therapeutic potential in various inflammatory diseases. However, its effect on asthma inflammation remains unclear. Methods: experiments-to investigate 3PO's anti-inflammatory action in asthma. Results: Firstly, the construction of a protein-protein interaction (PPI) network identified nine hub genes: TNF, IL6, BCL2, IL1B, CASP3, RELA, MAPT, MAOB, and MAOA. Then, the KEGG pathway enrichment analysis identified the nuclear factor kappa B (NF-κB) pathway as a pivotal target. Molecular docking and dynamics simulations suggested stable interactions between 3PO and key inflammatory markers, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and NF-κB P65. For experimental validation, the OVA-induced asthma mouse models were established. PFKFB3 expression was markedly upregulated in the lung tissues of asthmatic mice. 3PO treatment modulated mediator levels in BALF of asthmatic mice by reducing interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-13 (IL-13), interleukin-17A (IL-17A), IL-1β, TNF-α, IL-6, lactate, Immunoglobulin E (IgE), and chemokine (C-C motif) ligand 11 (CCL11), while elevating interleukin-10 (IL-10). 3PO treatment alleviated hallmark features of asthma in this model, such as airway mucus hypersecretion, goblet cell hyperplasia, and peribronchial infiltration of inflammatory cells, particularly eosinophils. Furthermore, 3PO treatment reversed key protein changes in lung tissue by increasing inhibitor of nuclear factor Kappa B-α(IKB-α) and decreasing p-NF-κB P65. Discussion: These results indicate that 3PO exerts anti-inflammatory effects in asthma, and this process is associated with the suppression of the NF-κB pathway, providing a novel therapeutic rationale for asthma therapy.