Thesium chinense Turcz. and its compound astragalin alleviate lipopolysaccharide-induced acute lung injury via the PI3K/AKT/p53 signaling pathway

Abstract

Ethnopharmacological relevance

Thesium chinense Turcz. (TCT) is a traditional Chinese medicinal plant commonly used in clinical practice for respiratory diseases, known for its anti-inflammatory and antioxidant properties. However, the mechanisms by which TCT alleviates acute lung injury (ALI) remain not fully understood.

Aim of the study: This study aims to validate the efficacy and elucidate the mechanisms of TCT in the treating ALI through experimental and network pharmacology approaches.

Materials and methods

To find the bioactive substances in TCT, ultra-high-performance liquid chromatography in conjunction with quadrupole-exactive mass spectrometry (UHPLC-QE-MS) was employed. The therapeutic effects of TCT were assessed by oxidative stress assessments, histological analysis (HE staining), inflammatory cytokine evaluations (RT-qPCR, immunofluorescence, immunohistochemistry), and TUNEL apoptotic staining in mice with lipopolysaccharide-induced ALI models. TP53 was identified as a crucial therapeutic target and astragalin as a pivotal drug by network pharmacology analysis. Their binding properties were verified by molecular dynamics simulations using GROMACS. The effects of astragalin on cell apoptosis (flow cytometry), cytokine expression (RT-qPCR), and protein expression (Western blot) were examined in vitro using LPS-stimulated RAW264.7 macrophages.

Results

In TCT, 24 key bioactive chemicals were found using UHPLC-QE-MS. TCT significantly reduced LPS-induced ALI, decreased levels of inflammatory cytokines TNF-α, IL-6, and IL-1β, decreased NFκB expression, increased superoxide dismutase (SOD) activity, decreased malondialdehyde (MDA) content, and reduced lung tissue cell apoptosis, according to the pharmacological results. The PI3K/AKT/p53 signaling pathway is essential for treating ALI, and astragalin was identified as the main drug and TP53 as the critical target by network pharmacology analysis. Strong binding and stability between astragalin and TP53 were demonstrated by molecular docking and dynamics simulations. Astragalin dramatically decreased early and late apoptosis, blocked p-PI3K and p-AKT activation, activated p53, and repressed the production of TNF-α, IL-6, and IL-1β in vitro.

Conclusion

This study suggests that TCT and its active compound astragalin alleviate LPS-induced ALI by modulating the PI3K/AKT/p53 signaling pathway, offering a potential therapeutic approach for treating ALI.