Abstract / Summary
Introduction: Acute kidney injury (AKI) is a life-threatening condition associated with oxidative stress, inflammation, apoptosis, and disruption of renal cellular homeostasis. Glycitein is a bioactive isoflavone that has various biological activity, but little is known about its possible protective effects and underlying molecular processes in AKI caused by cisplatin (CP). Thus, combining network pharmacology, molecular docking, and experimental validation, study examined the possible protective function of glycitein against CP-induced kidney injury. Materials and methods: The databases SwissTargetPrediction, SuperPred, and Traditional Chinese Medicine Systems Pharmacology (TCMSP) were used to find targets linked to glycitein, while DisGeNET and GeneCards provided targets related to AKI. A Venn diagram was used to find overlapping targets, which were then examined using Cytoscape and STRING's compound-target and protein-protein interaction (PPI) networks. The CytoHubba plugin and the maximal clique centrality algorithm were used to identify hub genes. DAVID was used for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment studies. Molecular docking was used to assess interaction between glycitein and hub proteins. NRK-52E renal tubular epithelial cells treated with CP were used for experimental validation. CP (30 μM) was administered to the cells alone or with glycitein (10, 20, and 30 μM), with apigenin (30 μM) as the positive control. We used MTT, GSH/SOD, and MDA assays, ELISA, qRT-PCR, western blotting, immunofluorescence, TUNEL, and H2DCFDA-based fluorescence analysis to measure cell viability, oxidative stress, lipid peroxidation, inflammatory biomarkers, apoptosis, gene expression, protein expression, and intracellular ROS levels respectively. Result: Glycitein and AKI were found to have overlapped potential targets by network pharmacology, and PPI analysis indicated several major hub genes, such as PIK3R1, HIF1A, ESR1, EGFR, NFKB1, BCL2, NFE2L2, and MAPK1. According to GO and KEGG enrichment studies, these targets were connected to signalling pathways and biological processes related to oxidative stress, inflammation, apoptosis, fibrosis and cellular responses. The result of molecular docking showed that glycitein and the chosen hub proteins interacted favourably, with binding energies falling within the predetermined range of ≤ −5 kcal/mol. Glycitein administration was further assessed for its impact on oxidative stress, inflammatory mediators, apoptotic responses, and kidney injury-associated markers, such as KIM-1, IL-18, Nrf2, and NF-κB, in the CP-treated NRK-52E cells line. Conclusion: The findings indicate that glycitein may protect against CP-induced renal injury via a multi-target mechanism involving the regulation of oxidative stress, inflammatory signalling, and apoptosis. The integrated approach of network pharmacology, molecular docking, and in vitro experiments offers insights into glycitein's role in CP treated-AKI and suggests the need for further in vivo studies.