Therapeutic Role and Potential Mechanism of Astragalus Membranaceus (Huangqi) and Radix Paeoniae Rubra (Chishao) in Idiopathic Pulmonary Fibrosis – Network Pharmacology and Experimental Validation

Abstract

Idiopathic pulmonary fibrosis (IPF) is a chronic progressive fibrotic disease with unclear etiology and no effective treatment. This study aims to reveal the pathogenetic mechanism networks of multiple targets and pathways of IPF. Extract and metabolites of Astragalus membranaceus (AM) and Radix paeoniae rubra (RPR), two well-known traditional Chinese medicine have been proven to be effective in IPF. However, the underlying mechanisms of AM and RPR in remain unclear. Based on network pharmacology analysis, differentially expressed genes (DEGs) of IPF were obtained from the GEO database. Targets of Astragalus membranaceus and Radix paeoniae rubra were identified using TCM Systems Pharmacology Database and Analysis Platform and SwissTargetPrediction. Subsequently, a protein-protein interaction (PPI) network was built and analyzed using the STRING database and Cytoscape software. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Gene and Genomes (KEGG) analysis were performed using Metascape. Further, a component-target-pathway network and a Sankey diagram were used to obtain main active components and molecular docking was performed between the key active components and proteins encoded by key targets. Finally, in vivo studies were carried out based on network pharmacology. 117 common targets between DEGs of IPF and targets of drugs were screened out and included in the PPI network, in which AKT1, MAPK3, HSP90AA1, VEGFA, CASP3, JUN, HIF1A, CCND1,PTSG2 and MDM2 were predicted as the key targets. 117 targets were enriched in PI3K-AKT pathway, HIF-1 signaling pathway, apoptosis and MicroRNAs in cancer. Astragaloside III, (R)-Isomucronulatol, Astragaloside I, Paeoniflorin and β-sitosterol were selected as the main active components. The docking scores ranged from − 4.7 kcal/mol to -10.7 kcal/mol, showing a good binding affinity between main active compounds and key targets. In vivo studies indicated that AM and RPR ameliorated pathological lung fibrotic damage caused by bleomycin and reduced mRNA level of AKT1, HSP90AA1, CASP3, MAPK3 and VEGFA. In conclusion, this study identified AM and RPR as potential therapeutic agents for IPF via regulating AKT1, HSP90AA1, CASP3, MAPK3 and VEGFA.