Dibutyryl Cyclic AMP Attenuates Cerebral Ischemia-reperfusion Injury by Inhibiting Cuproptosis: A Preliminary Study

INTRODUCTION: Cerebral ischemia-reperfusion injury (CIRI) poses a significant challenge in the treatment of ischemic stroke. Dibutyryl cyclic AMP (dBcAMP), a cell-permeable cAMP analog, has previously been shown to exert therapeutic effects in CIRI, indicating its neuroprotective potential. However, its underlying mechanisms remain incompletely understood. METHODS: We employed an integrated approach. First, an unbiased RNA-sequencing analysis of hippocampal tissues from a murine model of CIRI (induced by unilateral common carotid artery occlusion, UCCAO) was conducted to generate hypotheses. Subsequently, the hypothesis was functionally assessed in vitro using HT22 hippocampal neuronal cells subjected to oxygenglucose deprivation/reperfusion (OGD/R). Key features of cuproptosis, including intracellular copper accumulation, mitochondrial membrane potential, and cell viability, were assessed. RESULTS: Transcriptomics revealed significant suppression of the cuproptosis pathway by dBcAMP. Functional experiments confirmed that dBcAMP treatment significantly reduced OGD/R-induced intracellular copper accumulation (p < 0.05), restored mitochondrial membrane potential (p < 0.05), and improved neuronal survival (p < 0.05). DISCUSSION: These integrated findings suggest that dBcAMP may attenuate CIRI, at least in part, by inhibiting cuproptosis-a newly defined copper-dependent cell death pathway. This preliminary evidence positions dBcAMP as a potential modulator of cuproptosis, revealing a therapeutic dimension beyond classical programmed cell death. CONCLUSION: This study provides initial evidence that dBcAMP-mediated neuroprotection involves the reduction of intracellular copper overload and preservation of mitochondrial integrity, pointing to cuproptosis inhibition as a promising mechanism for future therapeutic exploration.