Human Neural Progenitor Cell-Derived Exosomes Deliver miR- 100 - 5p Targeting NOX4 mRNA to Alleviate Oxidative Stress in Acute Ischemia Injury

The prevention and treatment of acute ischemic stroke have been longstanding challenges. Therapies targeting angiogenesis hold promising potential for ischemic injury repair. Reactive oxygen species (ROS) production, induced by the overexpression of NADPH oxidase 4 (NOX4), is a key factor that inhibits angiogenesis during the acute phase of ischemia/reperfusion. Therefore, targeting NOX4 gene expression can reduce ROS production and promote angiogenesis. In this study, human neural progenitor cell (hNPC)-derived-exosomal miRNAs were identified using high-throughput sequencing and online database, with miR-100-5p showing potential to suppress NOX4 expression. We then incorporated exogenous miR-100-5p into hNPC-derived exosomes through electroporation. These miR-100-5p-loaded exosomes were then applied to in vitro models of mouse brain microvascular endothelial cells (bEND.3) subjected to oxygen glucose deprivation and reperfusion (OGD/R). The results showed that miR-100-5p could significantly reduce NOX4 expression and ROS levels induced by OGD/R in bEND.3 cells. Similarly, in vivo analysis showed that mice models of middle cerebral artery occlusion (MCAO) injected with hNPC-derived exosomes loaded with miR-100-5p exhibited smaller brain infarct size, reduced apoptosis, and improved neurological performance compared to MCAO mice receiving PBS. Our findings demonstrate the successful delivery of miR-100-5p via hNPC-derived exosomes and its protective effect on brain microvascular endothelial cells following ischemia injury.