Delayed administration of Jiuwei Qianghuo Decoction combined with Zhuye Shigao Decoction mitigates the severity of influenza A virus infection by inhibiting cell death and alleviating the subsequent cytokine storm response.

ETHNOPHARMACOLOGICAL RELEVANCE: The high mortality rate associated with severe influenza partly results from delayed initiation of antiviral therapy and subsequent cytokine storms. Jiuwei Qianghuo Decoction combined with Zhuye Shigao Decoction (JZF) has been clinically prescribed to prevent the progression to a more severe illness in influenza treatment. However, the precise mode of action and active components have not yet been elucidated.

AIM OF THE STUDY: This study aimed to determine the therapeutic efficacy and potential mechanism of action of JZF in severe influenza virus infection and cytokine shock syndromes.

MATERIALS AND METHODS: A lethal mouse model of H1N1 infection, the synergistic effect of TNF-α and IFN-γ-induced cytokine storm syndromes, the activity evaluation of monomers, and the verification experiments of the PBMCs death pathway in patients with H1N1 infection were combined to study the effect of JZF. RNA sequence analysis, cytokine and chemokine analysis, flow cytometry, western blot, alveolar macrophage depletion, and immunofluorescence staining combined with mass spectrometry analysis revealed the potential mechanism and elucidated the active components of JZF.

RESULTS: JZF demonstrated the most effective therapeutic effect in mice lethally infected with H1N1, 48 h post-inoculation. Transcriptome sequencing analysis of the lungs revealed that the top two downregulated pathways following JZF administration were TNF-α signaling via NF-κB and the IFN-γ response. Consequently, JZF significantly suppressed the production of various proinflammatory cytokines and chemokines. Mechanistically, JZF inhibited H1N1-triggered cell death, encompassing pyroptosis, apoptosis, and necroptosis. Furthermore, JZF alleviated the severity of infection in mice lethally inoculated with H1N1 by increasing the number of alveolar macrophages, in line with the suppression of macrophage death observed in vitro. This suppression was mediated by TNF-α and IFN-γ through the STAT1/ZBP1 signaling pathway. Lastly, furanocoumarins were identified as the key compounds in JZF responsible for inhibiting TNF-α- and IFN-γ-induced macrophage death.

CONCLUSION: These findings indicate that JZF inhibits viral infection and cytokine-induced macrophage death, which could potentially benefit patients with H1N1 or other infectious diseases by reducing tissue damage.