Immunomodulatory effects of stem cell-derived extracellular vesicles on NLRP3 inflammasome activation in the brain after chronic ethanol exposure

NOD-like receptors (NLRs) and inflammasome complexes play critical roles in the neuroinflammatory responses triggered by chronic ethanol exposure. While our previous work demonstrated that mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) attenuate binge ethanol-induced NLRP3 inflammasome activation in the adolescent hippocampus, their broader effects on other NLR pathways and brain regions remains unclear. Here, we investigated the therapeutic potential of intravenously administered adipose-derived MSC-EVs (20 µg/dose every 10 days) in a murine model of chronic alcoholism (10% ethanol in drinking water for 3 months), focusing on their ability to modulate multiple inflammasome sensors (NLRP3, NLRC4, NLRP1, AIM2) and downstream effectors (caspase-1, caspase-11/4, IL-1β, IL-18) across the prefrontal cortex, hippocampus and striatum. qPCR analysis revealed that chronic ethanol exposure significantly upregulated the expression of these inflammasome-related components in all three brain regions, whereas MSC-EV treatment effectively suppressed their activation. Notably, MSC-EVs normalized ethanol-induced overexpression of inflammasome sensors and downstream effectors, indicating a broad attenuation of inflammasome-driven neuroinflammatory responses. These findings expand current understanding of MSC-EVs as a multifaceted therapy for ethanol-related neuropathology, highlighting their capacity to simultaneously mitigate diverse NLR inflammasome pathways across key brain areas involved in addiction and cognitive dysfunction.