Mechanisms of Okanin against wound healing based on network pharmacology, molecular docking and molecular dynamics simulation.

Wound healing is a critical aspect of modern medicine, impacting patient health, quality of life, and healthcare resource allocation. Okanin, a flavonoid from the Asteraceae family, has shown potential in promoting wound healing. This study investigates okanin's key molecular targets, binding affinity, and mechanisms of action using network pharmacology, molecular docking, molecular dynamics simulations, and in vivo experimental validation. Okanin's potential targets were identified using the Comparative Toxicogenomics Database (CTD) and SwissTargetPrediction, while wound healing-related targets were sourced from GeneCards and DrugBank. Overlap analysis of these datasets revealed common targets. Key target proteins were filtered through protein-protein interaction (PPI) analysis using the STRING database. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted using Metascape to build a drug-target-pathway-disease network. Molecular docking was performed with AutoDockTools, and binding affinity was evaluated through energy scores, particularly with AURKA and HDAC1. Molecular dynamics simulations with GROMACS confirmed the stability of okanin-target complexes. ADME/T properties were assessed using SwissADME and ProTox-3.0 to evaluate pharmacokinetics and toxicity. In vivo quantitative real-time PCR (qRT-PCR) was performed to assess the expression of selected target genes in a mouse wound model following topical okanin treatment. A total of 72 common targets were identified between okanin and wound healing. PPI network analysis highlighted 17 key targets, with molecular docking revealing the highest binding affinity for AURKA and HDAC1 (ΔG = - 8.8 kcal/mol for both). GROMACS were then run on the top complexes. Target-ligand stability was quantified by convergence of RMSD/Rg, sustained hydrogen-bond counts, and MM/GBSA binding free energies (AURKA, - 24.27 ± 3.65 kcal/mol; HDAC1, - 47.7 ± 1.60 kcal/mol), confirming robust interactions. SwissADME predicted good drug-likeness (MW = 288.25 g/mol; logP = 1.69; high GI and moderate skin permeability) and no P-gp liability, while ProTox-3.0 indicated low systemic toxicity (LD₅₀ = 2500 mg/kg). qRT-PCR results demonstrated that okanin treatment significantly downregulated AURKA and PIK3R1, while upregulating HDAC1, in wounded skin, supporting the predicted molecular interactions and regulatory functions. Okanin promotes wound healing through multiple molecular targets and pathways, including antioxidant, anti-inflammatory, and cell proliferation mechanisms. Its high binding affinity for AURKA and HDAC1, along with modulation of the IL-17 and AMPK signaling pathways, underscores its therapeutic potential. This study provides a comprehensive theoretical and experimental framework for the development of okanin as a topical agent for wound healing, with future research focusing on formulation development and translational applications.