Molecular and metabolic mechanisms of bufalin against lung adenocarcinoma: New and comprehensive evidences from network pharmacology, metabolomics and molecular biology experiment. (May 2023)

• Record Type: Journal Article
• Authors: Shi, Shulong; Zhao, Sihao; Tian, Xinchen; Liu, Fen; Lu, Xiulian; Zang, Hengchang; Li, Feng; Xiang, Longquan; Li, Luning; Jiang, Shulong
• Is Part Of: Computers in biology and medicine. Volume 157(2023)
• Publication Date: 2023-05
• Subjects: Bufalin -- Network pharmacology -- Metabolomics -- Lung adenocarcinoma -- Molecular biology; Medicine -- Data processing -- Periodicals; Biology -- Data processing -- Periodicals; 610.285
• DOI: 10.1016/j.compbiomed.2023.106777
• Languages: English
• ISSNs: 0010-4825

Abstract

Abstract: Background: This study aims to evaluate the efficacy and therapeutic mechanism of bufalin on lung adenocarcinoma (LUAD) through a comprehensive strategy integrating network pharmacology, metabolomics and molecular biology verification. Methods: The putative targets of bufalin were discerned from PharmMapper and Swiss Target Prediction database. LUAD-related targets were obtained by target filtering of GeneCard database and data mining of GEO database. PPI network was constructed to screen the core targets, and their clinical significance was assessed through several public databases. GO and KEGG pathway analyses were performed to identify possible enrichment of genes with specific biological themes. Molecular docking and molecular dynamics (MD) simulation were employed to determine the correlation and binding pattern between bufalin and core targets. The potential mechanisms of bufalin acting on LUAD, as predicted by network pharmacology analyses, were experimentally validated using in-vitro and in-vivo models. Finally, the effects of bufalin intervention on metabolite profile and metabolic pathway in LUAD nude mice were investigated by non-targeted metabolomics. Results: 209 bufalin targets and 1082 LUAD-associated targets were harvested, of which 51 intersection targets were identified. 10 core targets including Akt1, STAT3, EGFR, CASP3 and SRC were picked out through network topology analysis, and they had a potent binding activity with bufalin as indicated by molecular docking and MD simulation. Hub module of PPI network was closely related to cell proliferation and apoptosis. GO and KEGG enrichment analyses suggested that bufalin exerted therapeutic effects on LUAD possibly by inhibiting proliferation and promoting apoptosis via PI3K/Akt, FoxO1 and MAPK/ERK pathways, which were confirmed by a series of in-vitro studies as well as HE, TUNEL and Ki-67 staining of tumor tissues. Further metabolomics analysis revealed that bufalin mainly regulated ABC transporter and remodeled AA metabolism, thereby contributing to the treatment of LUAD. Conclusion: From molecular and metabolic perspective, the present study not only provided a unique insight into the possible mechanisms of bufalin against LUAD after successfully filtering out associated key target genes, differential endogenous metabolites, and signaling pathways, but also proposed a novel promising therapeutic strategy for LUAD. Graphical abstract: Image 1 Highlights: A novel and comprehensive interpretation integrating network pharmacology, molecular biology validation, and metabolomics. The combination of molecular docking and MD simulation. The combined application of DEG analysis and WGCNA algorithm greatly improved the discrimination ability of LUAD targets. In-vitro and in-vivo experimental verification. Provide a reliable direction on the molecular design of the novel anti-LUAD agents.