Globally, ischemic stroke (IS) continues to rank among the leading causes of permanent disability. Globally, around 15 million individuals struggle with stroke annually, and incidence has increased over time since 1990–2017 (Ahmed et al., 2025). Although recent decades have seen a decline in stroke mortality and incidence, the total no of stroke cases, associated fatalities, and resulting global burden are still high. Despite a huge burden only tPA is approved with a limited therapeutic window and stringent contraindications, thrombolytic therapy (Liu et al., 2021). Currently, the ideal strategy for stroke therapy is still controversial. Nevertheless, oxidative stress-associated mitochondrial damage after ischemia contributes to neurological deficit. Although various neuro-protective compounds have been demonstrated promising results in preclinical rodent models of IS, but their translation into effective clinical therapies has been largely unsuccessful (Farina et al., 2021). Therefore, there is an urgent need to find novel preventive and therapeutic strategies for IS.

The reperfusion following ischemic stroke initiates a cascade of multifactorial mechanistic pathways. Among these, oxidative stress-mediated mitochondrial dysfunction has emerged as a critical therapeutic target in ischemic stroke (IS) treatment (Jurcau, 2022). The mitochondrial oxidative stress result a series of structural and functional disturbances, including damage to electron transport chain (ETC), augmented mitochondrial penetrability, swelling, release of pro-apoptotic proteins, and impairment of mitochondrial oxidative phosphorylation. These events disrupt ionic homeostasis and neurological function, ultimately leading to causes cerebral apoptosis and necrosis (Yang et al., 2015). Thus, protecting mitochondria from oxidative damage may represent a promising strategy for alleviating cerebral ischemic impairment. Hypoxia-inducible factor (HIF) and mitochondrial transcription factor A (TFAM) play crucial roles in maintaining mitochondrial homeostasis and promoting neuro-protection during ischemic stroke. However, excessive HIF-1 activation can elevate ROS production, destabilize mitochondrial membrane, and impair oxidative metabolism, ultimately triggering apoptosis and neuronal loss (Mitroshina et al., 2021). Furthermore, HIF-1 regulates mitochondrial biogenesis by interacting with the key modulators of mitochondrial transcription factor A (TFAM), which is essential for maintenance and function of mitochondrial DNA. Disruptions in this regulatory network may result mitochondrial fragmentation, energy failure, and increased vulnerability to ischemic injury (Der Chen et al., 2011). Therefore, understanding the intricate relationship between HIF-1, TFAM, and mitochondrial dysfunction could pave the way for novel therapeutic strategies interventions to improve stroke outcomes.

The correlation between Nrf2 and mitochondrial oxidative stress is well established. Nrf2 play a crucial role in maintaining mitochondria function, regulating cellular redox reactions, and preserving normal proteins function (Chen et al., 2023). Nrf2 activation promotes the expression of HO-1 which contributes to mitochondrial protection by regulating the activity of oxidative enzymes, such as SOD, GSH-Px, and CAT, thereby combating oxidative stress. Additionally, Nrf-2 also regulates mitochondrial biogenesis and quality control by modulating mitochondrial transcription factors such as TFAM (Wang et al., 2024). Hence, during ischemic stroke, HIF-1, Nrf-2, and TFAM collaboratively regulate mitochondrial function and neuro-protection as HIF-1 facilitating metabolic adaptation and mitophagy, Nrf-2 enhances antioxidant defenses to counteract oxidative stress, and TFAM ensures mtDNA stability and promoting mitochondrial biogenesis (Fan et al., 2019)(Chang et al., 2023). However, impairment in these pathways may exacerbate neuronal injury induced by ischemic stroke. Therefore, targeting Nrf2/HO-1/HIF-1/TFAM signaling axis and developing strategies to enhance mitochondrial function could offer promising therapeutic approaches for ischemic stroke.

Arbutin, a glycosylated hydroquinone, has antioxidant, anti-inflammatory, neuro-protective and anticancer activities (Boo, 2021). According to reports, AR suppressed expression of inducible nitric oxide (iNOS) and endorsed the nuclear factor erythroid 2-related factor 2 (Nrf-2) and Heme oxygenase-1 (HO-1) signaling pathways (Ashrafpour et al., 2024);(Okkay et al., 2024);(Alruhaimi, 2023). Moreover, AR showed neuroprotection by decreasing inflammatory markers i.e., IL-1β and TNF-α and enhancing neurotrophic markers in epilepsy (Ahmadian et al., 2019), IS (Kumar et al., 2021) and Parkinson's disease (Zhao et al., 2021). AR also demonstrated efficacy in alleviating neuro-inflammation and demyelination through reducing oxidative stress as well as activating astrocytes (Ebrahim-Tabar et al., 2020). Moreover, in models of Parkinson's disease (PD),AR also enhanced mitochondrial and motor function, potentially via its impact on oxidative stress and mitochondrial membrane potential (Ding et al., 2020). In Alzheimer's disease (AD) models, AR decreased MDA and nitrite concentration in rat brains subjected with streptozotocin (Dastan et al., 2019). However, arbutin's neuroprotective significance is not explored against the Nrf-2/HO-1/HIF-1α/TFAM pathway, which is involved in the complex pathology of ischemic stroke. The current study aimed to target the pathway above to treat oxidative stress-mediated mitochondrial dysfunction and neuronal death in ischemic stroke. Moreover, in silico analysis was also employed to elucidate the biological interactions of arbutin with Nrf2.