The network pharmacology prediction and experiment validation of Astragalus membranaceus for alleviating silicosis fibrosis via decreasing MMP9 and EGFR expression

To investigate the molecular mechanism of Astragalus membranaceus (AM) in alleviating silicosis fibrosis using bioinformatics and in vitro experiments. Active ingredients of AM and their corresponding targets were retrieved from the TCMSP and SwissTargetPrediction databases. These ingredients were screened based on oral bioavailability (OB ≥ 30%), drug-likeness (DL ≥ 0.18), and Lipinski's Rule of Five, which serves as a standard for identifying drug-like molecules with favorable pharmacokinetic properties in network pharmacology. Silicosis-related targets were collected from DisGENET, GeneCards, and the Comparative Toxicogenomics Database (CTD). Common targets were identified for protein-protein interaction (PPI) network analysis, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. Core ingredients and targets were selected through topological analysis in Cytoscape and subsequently validated using molecular docking (AutoDock), immunofluorescence and RT-qPCR assays. Eight main active ingredients of AM were identified, along with 236 AM targets and 7,706 silicosis targets, resulting in 113 shared targets. Further screening revealed three key ingredients: Quercetin, isorhamnetin, and kaempferol, as well as seven core targets: Akt1, CASP3, PTGS2, MMP9, ESR1, JUN and EGFR. The primary pathways involved included PI3K-AKT, IL-17, and MAPK. Molecular docking analyses indicated that MMP9 and EGFR exhibited higher binding energies with Quercetin. In vitro immunofluorescence results demonstrated that Quercetin downregulated the expression of MMP9, EGFR, and α-SMA in silicotic fibroblasts. Additionally, qRT-PCR assays confirmed that the Quercetin alleviated SiO₂-induced fibrotic process by suppressing PI3K-AKT pathway in the anti-fibrotic process. Quercetin, the key active ingredient of AM, exerts its anti-silicosis effect by inhibiting the expression of EGFR, MMP9 and the PI3K/Akt pathway.

Astragalus membranaceus; Molecular docking; Network pharmacology; Quercetin; Silicosis.