MiR-24-3p delivered by adipose derived stem cells exosomes alleviate macrophage pyroptosis via NLRP3/Caspase1/GSDMD in sepsis ALI
Wen Yin, Peng Wang, Hao Zhang, Xianqi Wang, Haian Fu, Jin Li, Ji‐Jun Chen, Peng Zhao, Wenjie Cheng, Yuhang Wang, Ruizhi Wang, Rui Zheng, Dahai Hu, Junjie Li
Abstract
BACKGROUND: Sepsis is a life-threatening condition associated with high morbidity and mortality, frequently resulting in acute lung injury (ALI) as one of its most severe complications. The pathogenesis of sepsis-induced ALI is significantly influenced by excessive inflammation, particularly that mediated by macrophages, plays a critical role in the pathogenesis of sepsis-induced ALI. This study sought to explore the protective effects and underlying mechanisms of adipose-derived stem cell exosomes (ADSCs-exo) on macrophage-mediated inflammation and pyroptosis. METHODS: ADSCs-exo were isolated and characterized using nanoparticle tracking analysis (NTA), Western blotting, and transmission electron microscopy. A macrophage pyroptosis model was established through lipopolysaccharide (LPS) stimulation. The effects of ADSCs-exo on the secretion of inflammatory cytokine and pyroptosis-related markers (NLRP3, Caspase1, GSDMD) was evaluated. Mechanistic studies included dual-luciferase reporter assays and NLRP3 inhibition experiments. In vivo, a cecal ligation and puncture (CLP) sepsis model was utilized to evaluate the impact of ADSCs-exo on pulmonary inflammation, histopathological damage, survival rates, macrophage polarization (M1/M2), and components of the pyroptosis pathway (NLRP3, Caspase1). RESULTS: Isolated ADSCs-exo displayed characteristic features of exosomes. The administration of ADSCs-exo mitigated LPS-induced hyperinflammation in macrophages, which was associated with a decreased expression of pyroptosis executors, namely Caspase1, GSDMD and NLRP3. Mechanistically, ADSCs-exo facilitated the delivery of miR-24-3p, which specifically targeted NLRP3 to inhibit pyroptosis. In mice subjected to CLP-challenge, the administration of ADSCs-exo improved lung pathology, enhanced survival rates, and reduced markers of pyroptosis in pulmonary tissues. CONCLUSIONS: Our findings indicate that ADSCs-exo attenuates ALI by modulating macrophage pyroptosis through the miR-24-3p/NLRP3/Caspase1/GSDMD axis, thereby presenting a promising therapeutic approach for sepsis-associated ALI.