Network Toxicology, Molecular Docking, and Molecular Dynamics to Explore the Epilepsy Mechanism Induced by Tetramine Poisoning.

Abstract

Tetramine poisoning commonly leads to epilepsy, with complex underlying mechanisms and poor treatment outcomes. This study aimed to explore the mechanisms of epilepsy induced by tetramine poisoning via network toxicology and molecular docking approaches. Tetramine poisoning targets were obtained from the SuperPred database, whereas epilepsy-related gene targets were identified through the GeneCards and OMIM databases. The intersection of tetramine targets and epilepsy-related genes revealed candidate targets. A proteinprotein interaction (PPI) network for these candidate targets was constructed via the String platform, after which the core functional modules were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted via the Metascape platform. Additionally, a targetpathway network involving Tetramine and brain injury was constructed via Cytoscape 3.8.2 software, enabling network topology analysis and screening of key components and targets. The results found that a total of 118 candidate targets for tetramine-induced epilepsy were identified. Pathway enrichment analysis indicated that tetramine-induced epilepsy is likely associated with the HIF-1, PI3K-Akt, Ras, Toll-like receptor, chemokine, and neurotrophin signaling pathways. The core targets include SRC, STAT3, HSP90AB1, MMP9, and HIF1A. To verify the above findings experimentally, this study established an epilepsy rat model by intragastric administration of different doses (0.1, 0.25, 0.5mg/kg) of tetramine. The seizure behavior was evaluated using the Racine scoring system, and hippocampal tissues were collected for subsequent tests. Real-time fluorescence quantitative PCR and Western Blot were used to detect the mRNA and protein expression levels of five core targets, as well as the phosphorylation levels of STAT3, SRC, Akt, and ERK. Immunofluorescence staining was used to observe the expression and distribution of p-STAT3 and HIF1A in brain tissues, and HE staining was used to assess the histopathological changes. The results showed that tetramine induced severe epileptic seizures in a dose-dependent manner. At the same time, the mRNA and protein expression levels of core targets HIF1A, MMP9, HSP90AB1, SRC, and STAT were all upregulated, and the phosphorylation levels of STAT3, SRC, Akt, and ERK were increased. Immunofluorescence and HE staining further confirmed the protein activation and pathological changes induced by tetramine.Network toxicology methods suggest that tetramine may induce epilepsy through multiple targets and signaling pathways. The above experimental results have preliminarily verified the key targets for network toxicology identification. However, the specific mechanism still needs to be further studied and confirmed.