N6-methyladenosine-modified miR-873 promotes receptor-interacting protein kinase 3-mediated necroptosis after intracerebral hemorrhage in mice and HT22 cells.

Abstract

BACKGROUND: Intracerebral hemorrhage (ICH) causes a severe form of stroke characterized by high morbidity, mortality, and long-term disability. Neuronal cell death is influenced at the posttranscriptional level. Certain microRNAs influence neuronal cell death at the posttranscriptional level by regulating receptor-interacting protein kinase 3 ( RIPK3 ), a key mediator of necroptosis. The specific mechanism by which miR-873 mediates neuronal necroptosis following ICH remains unclear. Epigenetic abnormalities, particularly N6-methyladenosine (m6A) modification, are increasingly recognized as critical contributors to ICH pathophysiology. METHODS: ICH mice model was established, followed by intracerebroventricular injection for gene manipulation. Brain water content was measured to assess cerebral edema. Neurological function was evaluated using the Morris water maze and neurological deficit scoring. Molecular and cellular analyses included Western blotting, quantitative real-time PCR, immunofluorescence, and luciferase reporter assays. Primary neuronal cultures, plasmid construction, and m6A RNA methylation quantification were performed to investigate underlying mechanisms. Differential gene expression was analyzed using microarray profiling, and data were statistically evaluated with appropriate analytical methods. RESULTS: The m6A modification is upregulated and positively involved in the functional role of miR-873 in ICH. miR-873 rescued necroptosis in ICH. miR-873 targets RIPK3 . RRACH (R = G or A; H = A, C, or U) m6A sequence motifs predominantly contribute to the m6A modification of miR-873 . The m6A modification regulates necroptosis in ICH. Knockdown of methyltransferase-like 3 improved the neurological function prognosis of ICH in mice. CONCLUSION: m6A modification modulates miR-873 expression, thereby influencing RIPK3-mediated necroptosis in ICH. These findings provide potential therapeutic targets for mitigating neuronal injury after hemorrhagic stroke.