Targeted gene delivery and enzyme inhibition in vascular and perivascular cells promote engraftment of transplanted parenchymal cells for lung and liver repair.

Idiopathic pulmonary fibrosis (IPF) is a chronic and irreversible interstitial lung disease characterized by progressive scarring of the lungs. The available therapeutic strategies are limited and primarily focus on slowing disease progression rather than achieving fibrosis reversal. Cardamonin (CDN), a food-derived natural chalcone, has exhibited anti-fibrotic activity in liver and kidney fibrosis models; however, its role and underlying mechanism in IPF remain unelucidated. Herein, we integrated network pharmacology, machine learning, molecular simulations, and in vitro experiments. Network pharmacology identified 135 overlapping targets between CDN and IPF, which demonstrated a significant enrichment in the Phosphatidylinositol 3-Kinase/Protein Kinase B signaling pathway (PI3K/AKT). Machine learning further prioritized 6 core targets, with IGF1 emerging as a key candidate. Molecular docking revealed a favorable binding energy of −7.9 kcal/mol for the CDN-IGF1 complex. Subsequent 100 ns molecular dynamics simulations further confirmed its robust binding stability, yielding a mean binding free energy of −150.978 kcal/mol. In vitro, CDN significantly mitigated fibrosis in bleomycin (BLM)-challenged A549 cells, downregulating the expression of α-smooth muscle actin (α-SMA) and fibronectin. This effect was accompanied by a beneficial reversal of epithelial–mesenchymal transition (EMT), as indicated by increased E-cadherin levels and decreased vimentin expression. Mechanistically, CDN significantly suppressed the IGF1/PI3K/AKT axis; this inhibitory effect was partially reversed by exogenous IGF1 supplementation and further enhanced by the PI3K-specific inhibitor LY294002. This work provides the evidence that CDN alleviates BLM-induced pulmonary fibrosis by targeting the IGF1/PI3K/AKT-EMT axis. These findings lend support to a robust mechanistic basis for developing CDN as a potential therapeutic candidate for IPF. It should be noted that these conclusions are drawn from in vitro experiments using A549 cells, and further validation in primary alveolar epithelial cells and animal models is warranted to confirm their physiological relevance.

Objective: Silicosis remains a debilitating occupational lung disease with limited therapeutic options, despite emerging evidence supporting pirfenidone’s (PFD) anti-fibrotic efficacy in clinical practice. However, the molecular circuitry governing PFD’s therapeutic actions in silicosis remains incompletely mapped, hindering mechanism-driven therapeutic optimization. To bridge this knowledge gap, we executed network pharmacology to replenish its molecular mechanisms and potential therapeutic targets.Materials and methods: We replicated a silicosis C57BL6/J mouse model and evaluated inflammation and fibrosis using HE, Masson, and Sirius Red staining assays. The expression of fibrotic markers α-SMA and Fibronectin were determined by immunofluorescence assay. Network pharmacology and molecular docking were used to predict potential therapeutic mechanisms and targets. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunofluorescence experiments were verified as the key predicted targets.Results: PFD alleviated the level of inflammation and collagen deposition and fibrotic markers α-SMA and Fibronectin expression in silicosis lung. Network pharmacology analysis predicted three potential target proteins, including TNF, MMP9, and NF-κB1, as well as ten possible signaling pathways. Molecular docking showed a good binding activity between PFD and hub genes. qRT-PCR and immunofluorescence confirmed that PFD inhibited TNF, MMP9, and NF-κB activation. Additionally, we found increased expression of TLR2, a key upstream gene of NF-κB.Conclusion: In conclusion, we identified TNF, MMP9, NF-κB1 and TLR2, that contribute to the therapeutic effects of PFD in silicosis. Mechanistically, PFD appears to mitigate silicosis pathogenesis through suppression of epithelial TLR2/NF-κB pathway activation.

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.