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

Abstract

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.