Ginsenosides Alleviate the Severity of Febrile Seizures via Targeting Dynamin‐Related Protein 1 to Promote Cortical Adenosine Elevation and Suppress the Cyclic Adenosine Monophosphate Signaling Pathway

Febrile seizure is a common pediatric neurological emergency that may potentially increase the risks of epilepsy and neurodevelopmental disorders. Ginsenosides are the primary active component of ginseng, demonstrating notable neuroprotective effects. However, the effects and mechanisms of Ginsenosides in febrile seizures remain poorly understood. This study aims to investigate the effects and potential molecular targets of Ginsenosides in mitigating febrile seizures and further elucidate its underlying mechanism. Seizure behaviors and EEG recordings were conducted in mice to investigate the effect of Ginsenosides on febrile seizures. Direct targets of Ginsenosides were identified through Thermal Proteome Profiling (TPP), subsequently validated by Surface Plasmon Resonance (SPR), Cellular Thermal Shift Assay (CETSA), and molecular docking. The mechanisms of Ginsenosides targeting Dynamin-related protein 1 (Drp1) in inhibiting febrile seizures were elucidated through proteomic analysis, molecular biology techniques, mitochondrial function assessments, and metabolite profiling. These findings demonstrate that Ginsenosides significantly attenuated febrile seizure severity and reduced the incidence of generalized tonic-clonic seizures (GTCS), showing a superior safety and efficacy profile compared to Ilepcimide. Using the TPP method, we identified and validated Drp1 as a promising direct target for therapeutic intervention in febrile seizures. Mechanistically, Ginsenoside-mediated Drp1 inhibition restored mitochondrial calcium homeostasis, promoting ATP production and elevating region-specific cortical levels of the endogenous anti-seizure metabolite adenosine. Moreover, Ginsenosides upregulated adenosine A1 receptor expression and suppressed the cyclic Adenosine Monophosphate (cAMP) signaling pathway, ultimately exerting anti-seizure effects. In summary, this study reveals that Ginsenosides significantly inhibit febrile seizures by directly targeting Drp1, thereby increasing adenosine levels and suppressing cAMP signaling, effectively suppressing seizures. Our findings demonstrate the potential of Ginsenosides in febrile seizures prevention and highlight Drp1 as a promising therapeutic target, thereby providing novel strategies for identifying targets of bioactive compounds.