Systems-level analysis uncovers eight hub genes and a resveratrol-based therapeutic strategy for ischemic stroke

Ischemic stroke is a leading cause of mortality and long-term disability worldwide, yet its underlying molecular mechanisms remain incompletely understood. The pathology involves complex interactions between vascular injury, immune dysregulation, metabolic imbalance, and mitochondrial dysfunction. Traditional single-gene or single-dataset approaches have limitations in capturing the multifactorial nature of the disease. We developed a comprehensive systems-level bioinformatics framework integrating five publicly available transcriptomic datasets (GSE119121, GSE146882, GSE22255, GSE37587, GSE58294). Differential expression analysis, protein-protein interaction network construction, functional enrichment analysis, gene-drug interaction mapping, and molecular docking simulations were performed. Eight core genes were identified through multi-dataset integration. Experimental validation was conducted using a transient middle cerebral artery occlusion (MCAO) rat model, with resveratrol treatment at 10 mg/kg and 30 mg/kg. qPCR, Western blot, TTC staining, H&E, NIR-II imaging, MRI, and neurological deficit assessments were performed. Integrated analysis identified eight core genes (SPTBN1, ARHGAP39, ANKRD44, FBXL19, FAM171B, SNORD77, FITM2, FIBIN) consistently dysregulated across datasets, showing strong associations with neuroinflammation, mitochondrial dysfunction, lipid metabolism, and immune regulation. In the MCAO model, all eight genes were significantly dysregulated at 24 h post-ischemia. Resveratrol treatment (30 mg/kg) normalized expression by 50%-60% for all genes (SPTBN1: from 3.2-fold to 1.5-fold; ARHGAP39: from 2.8-fold to 1.3-fold; FAM171B: from 3.5-fold to 1.6-fold; SNORD77: from 0.4-fold to 0.8-fold; all p < 0.01). Resveratrol reduced infarct volume by 52%, improved neurological scores by 60%, suppressed microglial activation (Iba-1 intensity reduced by 45.2%, p < 0.01), preserved mitochondrial membrane potential (108% improvement, p < 0.01), and reduced apoptotic cell death. Molecular docking revealed strong binding affinity between resveratrol and target proteins (FITM2: -8.2 kcal/mol). This study identifies a novel eight-gene signature associated with ischemic stroke and demonstrates that resveratrol effectively modulates these genes through coordinated regulation of neuroinflammation, mitochondrial function, apoptosis, and metabolic pathways. The multi-target therapeutic profile of resveratrol, validated through both computational and experimental approaches, positions it as a promising candidate for ischemic stroke therapy. The systems-level framework established here provides a robust foundation for future biomarker development and therapeutic discovery in complex neurological disorders.