Neuroprotective effect of genistein nanoparticles on ischemic cerebral infarction.

Abstract

BACKGROUND: Ischemic stroke (IS) significantly impairs quality of life, yet current therapies are limited by narrow therapeutic windows and risks such as hemorrhagic transformation. Genistein (Gen) is a natural compound with anti-inflammatory and antioxidant properties capable of crossing the blood-brain barrier (BBB); however, its clinical application is hindered by poor solubility. This study aimed to develop genistein nanoparticles (Gen NPs) with reduced particle size and enhanced solubility, and investigate their neuroprotective effects against IS. METHODS: Gen NPs with improved solubility were synthesized using the emulsion-solvent evaporation method and characterized by mass spectrometry and electron microscopy. Neuroprotective efficacy was evaluated in a mouse middle cerebral artery occlusion (MCAO) model. Infarct volumes were quantified by TTC staining, and behavioral tests were conducted to assess sensorimotor function. Network pharmacology and molecular docking approaches were utilized to identify therapeutic targets related to apoptosis and inflammatory pathways. BBB integrity was assessed through Evans blue (EB) dye extravasation and IgG permeability assays. Neuronal preservation was confirmed by Nissl and Fluoro-Jade C (FJ-C) staining. RESULTS: Gen NPs exhibited rod-like nanostructures with enhanced solubility, smaller particle sizes, and improved uniformity. Experimental findings demonstrated that Gen NPs had superior efficacy compared to free Gen in reducing infarct volume and enhancing neurobehavioral outcomes, with an optimal therapeutic dose identified as 2 mg/kg. Additionally, Gen NPs modulated multiple pathological mechanisms, including apoptosis and inflammation, thereby preserving BBB integrity and attenuating neuronal injury. CONCLUSION: We successfully synthesized Gen NPs exhibiting improved water solubility, reduced particle size, and enhanced uniformity, demonstrating their superior therapeutic potential for IS treatment. These findings highlight the multifaceted neuroprotective mechanisms of Gen NPs, suggesting promising prospects for clinical translation.