Investigation of the effect and mechanism of Fei Re Pu Qing powder in treating acute lung injury (ALI) by modulating macrophage polarization via serum pharmacology and network pharmacology

Abstract

Ethnopharmacological relevance

Acute lung injury (ALI) is a clinical critical illness characterized by progressive pulmonary inflammatory response and high mortality. An increasing number of studies have confirmed that traditional Chinese medicine (TCM) and compound prescriptions hold considerable promise in the prevention and treatment of respiratory inflammatory diseases. Fei Re Pu Qing Powder (FRPQ) is an efficacious prescription used in clinical treatment of pneumonia in Tibet, China. Its potential effects and mechanisms on ALI are worthy of further study.

Aim of the study

This study aims to characterize the bioactive constituents of FRPQ and elucidate its potential target sites and underlying mechanisms of action in ALI.

Materials and methods

Firstly, the pharmacodynamic effects of FRPQ were assessed using an LPS-induced rat model of ALI. Subsequently, the optimal pharmacodynamic dose was determined for a time-course study to evaluate the onset and duration of FRPQ's pharmacological actions. Based on the identification of potential active components in FRPQ-containing serum using UHPLC-Q Exactive HFX, core components, key targets, and signaling pathways were systematically screened via network pharmacology approaches. Subsequently, molecular docking analysis was performed on the identified core components and key targets to validate their binding activity and affinity. Finally, the potential mechanism underlying its effect on ALI was further verified using the ALI rat model and the M1/M2 polarization model of NR8383 cells.

Results

FRPQ exerts rapid and long-lasting effects on ALI rats. 63 prototype components of FRPQ were identified in the drug-containing serum, among which 25 have potential activity. An active components - intersection targets - ALI network was constructed, and 6 core components (Kaempferol, Piperine, Phloretin, etc.) were selected. PPI analysis identified 9 targets closely related to the therapeutic effect of ALI. GO/KEGG analysis indicated that FRPQ treatment of ALI is closely related to multiple inflammatory signaling pathways. Molecular docking showed that the binding energy of the 6 core components with the 9 core targets was ≤ −5.4 kcal/mol, indicating strong binding affinity. In vivo experiments showed that FRPQ could significantly reduce macrophage infiltration in the lung tissue of ALI rats, effectively down-regulate the expression of NF-κB/p65 protein, and the changes in the transcriptional levels of inflammatory mediators in lung tissue and the content of inflammatory factors in alveolar lavage fluid further verified the regulatory effect of FRPQ on the NF-κB signaling pathway. In vitro experiments showed that the drug-containing serum significantly inhibited the polarization of NR833 cells to the M1 phenotype and promoted their polarization to the M2 phenotype. The inhibition of M1 polarization may be achieved through negative regulation of the NF-κB signaling pathway, providing potential mechanism support for the treatment of ALI.

Conclusions

This study systematically analyzed and preliminarily elucidated the active components of FRPQ and their potential mechanisms of action by integrating serum pharmacology with network pharmacology approaches. The findings indicate that FRPQ may contribute to the treatment of ALI through the regulation of the NF-κB signaling pathway and the modulation of macrophage M1/M2 polarization. This advancement offers critical experimental evidence and theoretical underpinnings for the further development and clinical application of FRPQ.