Yi-Fei formula mitigates influenza virus NS1-induced macrophage dysfunction by activating the NF-κB signaling pathway

Influenza virus nonstructural protein 1 (NS1) significantly contributes to immune evasion and macrophage dysfunction, which exacerbates the severity of viral pneumonia. The present work analyzed the therapeutic efficacy of Yi-Fei formula (YFF), a traditional Chinese medicine (TCM), in macrophage dysfunction induced by NS1 of the influenza virus and the associated molecular mechanisms. We characterized the primary active components of YFF by conducting ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). We established an NS1-overexpressing macrophage model and found that 40 μg/mL YFF significantly restored cell viability impaired by the overexpression of NS1. YFF enhanced the proliferation, migration, and survival of NS1-overexpressing macrophages while suppressing apoptosis by activating the NF-κB pathway. Notably, YFF effectively suppresses NS1-induced apoptosis and modulates the inflammatory response by upregulating key factors like IL-6 and TNF-α. Transcriptomic analysis underscored importants effect of NF-κB pathway on mediating YFF's immunomodulatory effects. These findings demonstrate that YFF can effectively attenuate NS1-induced macrophage dysfunction, shedding new lights on the molecular mechanisms related to YFF action, highlighting that it can effectively treat viral pneumonia by modulating the functions of macrophages.