Ethanol extract of Corydalis decumbens exerts neuroprotective effects against cerebral ischemia-reperfusion injury by targeting ALOX15 to suppress ferroptosis.

BACKGROUND: The management of ischemic stroke is complicated by cerebral ischemia-reperfusion (I/R) injury, presenting substantial obstacles for pharmaceutical advancement. Clinically, the rhizome of Corydalis decumbens (Thunb.) Pers., a traditional Chinese medicinal herb, is frequently employed for treating ischemic stroke. Prior studies have shown that its ethanol extract demonstrates a significant therapeutic impact in mitigating cerebral I/R injury. Nevertheless, the molecular mechanisms underlying the extract of Corydalis decumbens (CDE)'s therapeutic effects and its bioactive constituents remain incompletely characterized.

PURPOSE: The aim of this study is to elucidate the potential mechanism and bioactive ingredients of CDE in alleviating cerebral I/R injury.

METHODS: Evaluate the pharmacological activity of CDE on cerebral I/R injury through multiple pharmacological indicators. This study utilized integrated proteomics and metabolomics methods, combined with in vitro and in vivo molecular biology experiments, to elucidate the therapeutic mechanism of CDE on cerebral I/R injury. Furthermore, we used molecular docking and in vitro enzyme activity detection methods to search for the active substances of CDE acting on the target.

RESULTS: The research results indicate that CDE improves cerebral I/R injury by reducing neurological function scores, decreasing infarct volume, and increasing cerebral blood flow (CBF). Multi-omics joint analysis studies have demonstrated that CDE may mitigate the effects of cerebral I/R injury by modulating ALOX15-mediated ferroptosis. Utilizing the OGD/R cell model, we validated the regulatory influence of CDE on ferroptosis-related markers. Subsequent in vitro knockdown of ALOX15 or in vivo knockout experiments confirmed that the therapeutic efficacy of CDE against cerebral I/R injury was significantly reduced following ALOX15 depletion, with no observed improvement in ferroptosis-related indices. Finally, through molecular docking, in vitro enzyme activity detection, and cell experiments, we revealed that magnoflorine and corlumidine, principal constituents of CDE, exert neuroprotective effects through ALOX15 inhibition.

CONCLUSIONS: Our study demonstrates that the neuroprotective action of CDE and its bioactive compounds, magnoflorine and corlumidine, against cerebral ischemia-reperfusion injury is mediated by modulating ALOX15-driven ferroptosis, which establishes the basis for their clinical use in the treatment of ischemic stroke and offers a fresh viewpoint on the development of ischemic stroke clinical treatment.