Targeting the NEAT1/miR-23b-3p/Glutaminase Axis

Oxidative stress-induced cardiomyocyte injury constitutes a pivotal pathogenic mechanism driving heart failure progression. Emerging evidence implicates lncRNA NEAT1 in cardiovascular pathophysiology. This study delineates the mechanistic cascade through which NEAT1 orchestrates redox adaptation via glutamine metabolic reprogramming. Using H₂O₂-treated H9c2 rat cardiomyocytes as an oxidative stress model, we identified dose-dependent NEAT1 upregulation concomitant with miR-23b-3p suppression. NEAT1 silencing or miR-23b-3p overexpression exacerbated oxidative vulnerability, manifested by elevated viability loss and apoptosis rate. Bioinformatic prediction coupled with RNA pulldown and dual-luciferase assays established NEAT1/miR-23b-3p ceRNA interaction. Sublethal oxidative stress triggered adaptive glutaminolysis activation characterized by elevated glutamine uptake and GLS activity. Luciferase assay validated miR-23b-3p directly targeted GLS 3'UTR, while metabolic rescue assays demonstrated GLS reconstitution reversed miR-23b-3p-mediated metabolic suppression and restored oxidative tolerance. Epistatic analysis confirmed NEAT1's cardioprotection requires miR-23b-3p/GLS axis modulation, with miR-23b-3p restoration abrogating NEAT1-overexpression benefits. These findings were further corroborated in an in vivo rat model of myocardial ischemia-reperfusion injury, which recapitulated the upregulation of NEAT1 and GLS, downregulation of miR-23b-3p, and enhanced glutaminase activity in the infarct risk zone. Collectively, these results suggest the existence of a novel NEAT1/miR-23b-3p/GLS regulatory axis governing redox-stress adaptation in both cellular and animal models.