Stroke remains a leading cause of morbidity and mortality globally (Feigin et al., 2025; Prust et al., 2024), yet effective neuroprotective treatments are scarce. While animal studies have demonstrated the protective effects of remote ischemic preconditioning (RIPC) against ischemia-reperfusion injury, its translation to clinical stroke therapy remains limited (Beretta et al., 2025). Our team has demonstrated the protective effects of RIPC against stroke as early as in 2008 (Ren et al., 2008). Noninvasive RIPC methods, such as tourniquet-induced ischemia on the upper extremities, have shown promise for clinical application (Li et al., 2024a; Hou et al., 2022). RIPC achieves neuroprotection through various mechanisms, including reduced inflammation, attenuated oxidative stress, and modulation of signaling pathways like HIF-1α and AMPK, via various avenues, such humoral, nerves, and immune pathways (Gu et al., 2023; Jiang et al., 2024). However, a significant gap persists in understanding whether these protective mechanisms observed in animal studies can be translated to humans.

Critical questions remain regarding whether the mechanisms identified in animals can be extrapolated to human stroke patients. In the case of RIPC, these uncertainties are compounded by the complexity of stroke pathology (Goel et al., 2024), which varies across brain structures (McDonald et al., 2021; Liu et al., 2016), stroke subtypes, and populations of different genetic and ethnic backgrounds (Mishra et al., 2022). The protective mechanisms of RIPC likely involve region-specific effects within the brain and interactions with peripheral tissues (Bonova et al., 2020; Chen et al., 2018). Moreover, genetic and geographic variations may contribute to differences in RIPC efficacy, underscoring the need for population-specific and subtype-specific evaluations (Pico et al., 2020; England et al., 2019; Blauenfeldt et al., 2023; Chen et al., 2022). Understanding these factors is crucial for optimizing RIPC-based interventions for diverse patient populations and stroke types.

To address the translational challenge of linking RIPC-induced neuroprotection observed in animal models to human stroke risk, we designed a multi-level integrative study. First, we established a murine RIPC model and systematically mapped central and peripheral transcriptomic responses at distinct time points. Second, murine differentially expressed genes were translated to human orthologs and integrated with human eQTL data and multi-ancestry, multi-subtype stroke GWAS to identify RIPC-regulated beneficial genes (RBGs) using Mendelian randomization. Third, functional enrichment, colocalization, drug-target prediction, and epigenetic analyses were performed to characterize the biological relevance of these RBGs. Fourth, machine learning–based models were constructed to explore the association of RBG signatures with stroke-related genetic risk across populations. Finally, core RBGs were experimentally validated in the murine tMCAO model.

Importantly, while the genetic analyses link RBGs to human stroke susceptibility, the experimental validation in this study—based on short-term outcomes following tMCAO—primarily supports a role for these genes in enhancing resilience to acute ischemic injury and early recovery. Their potential contribution to true primary or secondary stroke prevention in humans remains inferential and will require validation in long-term clinical and interventional studies.