Polydatin mitigates seizures via mitochondrial protection: Mechanisms unraveled by network pharmacology and experimental validation

Abstract

This study investigated the antiepileptic effects of polydatin (PD) and its potential mechanisms. Racine scoring and EEG were used to assess seizure severity in a PTZ-induced mouse epilepsy model. The results demonstrated that PD significantly reduced the intensity of seizures. Behavioral tests revealed that PD improved learning, memory, and motor coordination. Histological analysis using HE and Nissl staining showed that PD alleviated hippocampal neuronal damage. TUNEL staining further confirmed that PD effectively prevented neuronal apoptosis. Network pharmacology predicted SIRT1 as a potential target of PD, and GO and KEGG pathway enrichment analyses indicated that PD regulated mitochondrial function. Therefore, PD may reduce neuronal damage by protecting mitochondrial function through SIRT1. This process was tested both in vitro and in vivo. In a Mg²⁺-free neuronal excitability model, CCK-8, LDH, and Hoechst staining results demonstrated that PD significantly increased cell survival and reduced apoptosis. Reduced mitosox staining, along with increased activity of Complexes IV and V and ATP production, indicated that PD mitigated mitochondrial function. Gene expression studies revealed that PD activated the SIRT1-PGC-1α signaling pathway via AMPK. In conclusion, PD exerted neuroprotective effects by regulating mitochondrial function, suggesting a potential therapeutic target for epilepsy.