Stroke is one of the most common causes of serious long-term disability and death in patients worldwide. It’s caused by an inadequate blood supply, which leads to brain damage. The globally accepted treatment strategy following cerebral ischemia is to ensure rapid restoration of blood flow and reoxygenation of ischemic tissues (Ding et al., 2024, Khoshnam et al., 2024). Unfortunately, when blood flow is restored, the brain suffers a secondary injury, also known as cerebral ischemia/reperfusion injury (CIRI), which is a complex pathological and physiological process involving energetic disturbances, increased release of excitatory amino acids, oxidative stress, inflammatory responses, and activation of apoptotic genes, ultimately leading to neuronal apoptosis or necrosis (Wang et al., 2024, Yang et al., 2025). Currently, the pathologic mechanisms that trigger CIRI are still being explored. Therefore, further clarification of the pathological mechanisms of CIRI, as well as the development of new therapeutic strategies for CIRI, are necessary and urgent.

Nuclear factor-erythroid 2 (NFE2)-related factor 1 (NRF1) is a transcription factor that contains a 43-amino acid cap “n” collar (CNC) motif and its adjacent basic region leucine zipper domain, belonging to the CNC-bZIP family (Ruvkun and Lehrbach, 2023). It is involved in multiple important biological processes such as redox signaling, cell metabolism, and proteasome regulation (Łuczyńska et al., 2024). Notably, NRF1 binds to the promoter regions of its target genes through antioxidant response elements (AREs), thereby driving the transcriptional activation of stress-resistant and cytoprotective genes, such as HMOX1, SOD1, or GCLC (Ruvkun and Lehrbach, 2023, Hamazaki and Murata, 2020). Additionally, studies have shown that oxidative stress, inflammatory response, and apoptosis are core processes of CIRI (Wang et al., 2024). The NRF1 and NRF2 pathways are the most important antioxidant stress systems, playing a crucial role in regulating apoptosis induced by oxidative stress (Hamazaki and Murata, 2020, Wang et al., 2022). Yang et al.’s report (Yang et al., 2024) indicated that during CIRI, downregulation of NRF1 promotes inflammatory response by regulating the expression of IL-6/TNF-α, leading to worsening of neurological deficits, increasing cerebral infarction volume, and exacerbating cortical histopathological damage. Ying et al.’s report (Yuan et al., 2023) showed that overexpression of NRF1 prevents CIRI by improving mitochondrial function. However, the molecular mechanism of NRF1 in CIRI is not fully understood.

N6-methyladenosine (m6A) modification is the most common internal modification in eukaryotic mRNAs, accounting for 80 % of RNA modifications. It affects RNA localization, nuclear export, translation, stability, and alternative splicing, playing a role in various important biological processes such as cell proliferation, differentiation, and apoptosis, and is closely related to the occurrence and development of various diseases (Bu et al., 2024, Pilala et al., 2025). m6A modification is reversible. Methyltransferases (such as METTL3, METTL4, METTL14, and WTAP) promote m6A modification of RNA, while demethylases (such as FTO and ALKBH5) remove m6A modification from RNA (Pilala et al., 2025). Reports have shown that m6A modification plays an important role in the progression of CIRI (Xu et al., 2024, Chen et al., 2025). Shao et al. found that total m6A levels significantly increased after CIRI, while the expression of fat mass and obesity-related protein (FTO) was downregulated (Shao et al., 2022). However, the specific mechanism by which FTO participates in CIRI is unclear.

In this study, we constructed a MCAO/R rat model and a OGD/R-induced neuronal cell model to mimic CIRI in vitro and in vivo, and investigated the potential mechanisms by which NRF1 regulates OGD/R-mediated neuronal apoptosis and its effects on CIRI in rats by NRF1 gain-of-function and loss-of-function experiments. Our study provides new perspectives for the development of therapeutic strategies for CIRI.