Integrated network pharmacology and transcriptomics analysis elucidated the underlying mechanisms by which Pegaeophyton scapiflorum alleviated LPS-induced acute lung injury.

ETHNOPHARMACOLOGICAL RELEVANCE

Acute lung injury is one of the most fatal lung diseases and has a significant impact on mortality and morbidity. Currently, ALI treatment options remain limited. Pegaeophyton scapiflorum (DHJ) has been documented in Dumu Materia Medica, as clearing heat from the lungs, and are clinically used for respiratory disorders. However, their therapeutic mechanisms in ALI are not yet fully elucidated.

AIM OF THE STUDY

The research aimed to integrate network pharmacology with transcriptomics to clarify the pharmacological effects and therapeutic impact of DHJ against LPS-induced ALI. Thus laying a solid foundation for regarding DHJ as possible intervention measures to improve ALI.

MATERIALS AND METHODS

The pharmacological mechanisms underlying the effects of DHJ on ALI were preliminarily explored through network pharmacology analysis. The efficacy of DHJ in LPS-triggered ALI mice was evaluated through histopathology and biochemical analysis. Transcriptomic profiling was performed on lung tissues from ALI mice to elucidate the potential mechanisms of action of DHJ. Furthermore, an in vitro ALI model using LPS-induced RAW264.7 mouse macrophages was developed. The protective pharmacological mechanisms of DHJ were further validated using immunofluorescence staining, qRT-PCR, and Western Blot analysis.

RESULTS

DHJ effectively ameliorated pulmonary edema in ALI mice, as evidenced by lung dry and wet specific gravity. The use of DHJ drugs attenuated LPS-induced alveolar structural damage, pulmonary hemorrhage, and inflammatory cell infiltration in mice, and HE staining demonstrated these results. Meanwhile, the high-dose group markedly reduced LPS-induced elevations in inflammatory markers (IL-6, IL-1β, TNF-α). Additionally, DHJ inhibited the generation of ROS in ALI mice. They inhibited the rise of MDA and MPO content and alleviated the decrease of SOD and GSH content caused by LPS stimulation. The results showed that DHJ had protective effects on LPS-induced ALI mice. Based on the identification of the chemical constituents of this herb in the pre-experimental period, network pharmacology predicted the potential mechanisms of DHJ against ALI. It was found that the above chemical components may act through TLR4/NF-κB/PI3K/AKT pathway. DHJ's protective impact on ALI mice was evident through reduced inflammation, immune response adjustment, and oxidative stress mitigation. Transcriptomics, western blotting, immunofluorescence, immunohistochemistry, qRT-PCR and molecular docking showed that DHJ inhibited TLR4 and MYD88 levels, as well as NF-κB, IκBa, PI3K, and AKT protein phosphorylation levels.

CONCLUSIONS

DHJ alleviated lung pathology in ALI mice, suppressed the LPS-induced elevation of inflammatory factors and oxidative stress enzymes, mitigated inflammation and oxidative stress, and modulated dysregulated immune responses to attenuate acute lung injury. These effects are mediated via TLR4/NF-κB/PI3K/AKT pathway. The elucidated mechanism offers a theoretical foundation for utilizing DHJ in the treatment of LPS-induced ALI.