An exploration of the protective effects of Ginsenoside Rb1 against acute lung injury using network pharmacology, molecular docking, molecular dynamics simulations, and in vivo experiments.

Background: Acute lung injury (ALI) is a severe inflammatory disorder where neutrophils contribute to inflammation and tissue damage by forming neutrophil extracellular traps (NETs). While Ginsenoside Rb1 (Gs-Rb1) has been shown to offer protective effects in ALI, it remains unclear whether its benefits in Lipopolysaccharide (LPS)-induced ALI involve modulation of NETs.

Objective: The study aimed to assess the protection of Gs-Rb1 against ALI using bioinformatics analyses and animal experiments.

Methods: Potential targets of Gs-Rb1 and ALI were identified through several databases and analyzed using protein-protein interaction (PPI) networks, GO and KEGG pathway enrichment, molecular docking, qRT-PCR, and molecular dynamics simulations to pinpoint key targets of Gs-Rb1. The compound's therapeutic effects were further explored in mouse models of LPS-induced ALI.

Results: A total of 90 proteins were identified as shared targets between Gs-Rb1 and ALI. The top 10 targets were selected based on degree values from PPI networks. GO and KEGG enrichment analyses revealed links to 306 biological processes, 29 molecular functions, 63 cellular components, and 148 signaling pathways, suggesting that NET formation plays a central role in the therapeutic effects of Gs-Rb1 on ALI. Molecular docking showed strong binding affinity between Gs-Rb1 and the core targets, while qRT-PCR confirmed significant changes in AKT1 expression following Gs-Rb1 treatment. Molecular dynamics simulations further supported the binding of AKT1 to Gs-Rb1. In LPS-induced mouse models of ALI, Gs-Rb1 treatment attenuated histological damage, reduced the wet/dry mass ratio, and lowered levels of TNF-α, IL-1β, and IL-6. Furthermore, it decreased the fluorescence intensity and protein expression of CitH3, NE, and MPO and downregulated the protein ratios of p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR.

Conclusion: These findings suggest that Gs-Rb1 may alleviate inflammation in ALI by inhibiting NET formation, likely through modulation of the PI3K/AKT/mTOR axis.