Therapeutic Targeting of the IRF9/RTN4/RHOA/ROCK Pathway via RVG29‐Modified PLGA Nanoparticles and rTMS for Neural and Vascular Regeneration Post‐Cerebral Infarction

Cerebral infarction, a leading cerebrovascular disease, often results in severe neurological impairments and high mortality. This study investigates a novel therapeutic approach involving small interfering RNA targeting Interferon Regulatory Factor 9 (si-IRF9) delivered by RVG29-functionalized poly(lactic-co-glycolic acid) nanoparticles (NPs) (RVG29-PNPs@si-IRF9), in combination with high-frequency repetitive transcranial magnetic stimulation (rTMS), in promoting post-stroke regeneration. Using a middle cerebral artery occlusion rat model and an in vitro oxygen-glucose deprivation/reoxygenation system, the regenerative efficacy of this combinatory therapy is evaluated on both neural and vascular recovery. Mechanistically, our results identify the IRF9/Reticulon 4 (RTN4)/Ras homolog family member A (RHOA)/Rho-associated coiled-coil containing protein kinase (ROCK) pathway as a key mediator, which is effectively inhibited by RVG29-PNPs@si-IRF9. This inhibition enhances neurogenesis and angiogenesis, particularly when combined with rTMS. Moreover, the NP system demonstrates excellent biocompatibility and targeted delivery, highlighting its potential as a therapeutic platform for stroke rehabilitation. These findings provide a new perspective on integrating nanotechnology and neuromodulation to facilitate functional recovery after cerebral infarction.