Regulation of Neuronal Pyroptosis Through NLRP3 by Delivering miR-22 Using Lipid Nanoparticles in Mice with Cerebral Ischemia-Reperfusion Injury

Abstract

Liposomes present a promising strategy for microRNA (miRNA) delivery, capitalizing on their unique properties to enable effective therapeutic interventions. In this study, we investigate lipid nanoparticles (LNPs) as carriers to delivery miR-22, aiming to mitigate neuronal pyroptosis by targeting nucleotide-binding oligomerization domain (NOD)-like receptor family pyrin domain containing 3 (NLRP3). In vitro, HT-22 cells were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) to assess cell viability, lactate dehydrogenase (LDH) levels, and pyroptosis. The pyroptosis-related protein expression was determined by Western blot analysis. The interaction between miR-22 and NLRP3 was assessed by dual luciferase assays. LNPs were employed to deliver miR-22 precursor oligonucleotides (LNP/miR-22) to HT-22 cells. miR-22 overexpression models were constructed to investigate its impact on OGD/R-induced pyroptosis. In vivo, a mouse model of cerebral ischemia-reperfusion was established to investigate the effects of LNP/miR-22 treatment, NLRP3 inhibitor (MCC950), or NLRP3 activator (Nigericin sodium salt). Neural damage and pyroptosis in the hippocampi were evaluated using staining techniques and immunofluorescence. The expression levels of pyroptosis-related proteins in the hippocampi were analyzed by western blotting. Results demonstrated that OGD/R reduced cell viability, increased LDH levels, and induced pyroptosis In vitro. NLRP3 overexpression exacerbated OGD/R-induced pyroptosis. miR-22 was found to target and downregulate NLRP3 expression, leading to reduced pyroptosis. In vivo, miR-22 overexpression suppressed NLRP3 activation, effectively attenuating pyroptosis. In conclusion, LNP-mediated delivery of miR-22 offers a promising strategy to alleviate neuronal pyroptosis by targeting NLRP3, holding potential for the treatment of cerebral ischemia-reperfusion injury.