Healthy young human plasma-derived exosomes enhance neural stem cell therapy by suppressing pyroptosis via TXNIP/NLRP3 after intracerebral hemorrhage.

BACKGROUND: Neural stem cells (NSCs) transplantation holds promise for intracerebral hemorrhage (ICH) treatment, but its efficacy is limited by poor survival and aberrant differentiation of grafted cells. Here, we demonstrate that exosomes derived from healthy young donor plasma, a natural nanomaterial protect NSCs against pyroptosis, a gasdermin-dependent inflammatory cell death process triggered by ICH. METHODS: Plasma exosomes were extracted from young (Y-exo) and old (O-exo) healthy individuals and characterized. An in vitro model of ICH was established by hemin treatment. For the in vivo study, the mouse ICH model was induced by autologous blood, a combined transplantation of Y-exo and NSCs was then performed as the therapeutic intervention. The protective effects of exosomes on NSCs were assessed via western blotting, immunofluorescence, ELISA, qPCR, and Calcein/PI detection. The therapeutic effects of combined transplantation of Y-exo and NSCs on ICH mice were evaluated through in vivo imaging systems and a series of behavioral tests. RESULTS: Exosomes derived from young plasma exert protective effects by supporting NSC survival, boosting their proliferative and differentiation capacity in vitro, and ameliorating the peri-hematoma microenvironment in vivo. Strikingly, the efficacy of Y-exo is superior to that of O-exo. Subsequent studies will use Y-exo, in vitro, the Y-exo exerted their protective effects by inhibiting the NLRP3/Caspase-1/GSDMD-mediated pyroptotic pathway and reducing the release of inflammatory cytokines. In vivo, co-transplantation of NSCs and Y-exo enhanced NSCs survival, proliferation, and beneficial differentiation toward neuronal and oliodendroglial lineage while attenuating pyroptosis of NSCs and peri-hematoma tissue. Behavioral tests indicated that mice in the co-transplantation group exhibited superior functional recovery. MiRNA sequencing identified miR-16-5p as a key mediator enriched in Y-exo, which targeted TXNIP to disrupt NLRP3 inflammasome activation. Genetic and pharmacological interventions confirmed that miR-16-5p/TXNIP/NLRP3 signaling pathway is essential for Y-exo's anti-pyroptotic effects. CONCLUSIONS: Our study elucidates a previously unidentified mechanism whereby Y-exo improve neurological outcomes by alleviating the peri-hematoma inflammatory microenvironment, suppressing pyroptosis in transplanted stem cells, and altering their differentiation fate. This study highlights the potential of synergistic strategy to optimize NSCs-based therapy for stroke by combining youth-derived factors, offering new insights into regenerative therapeutics for neurological disorders.