Abstract
Objective: Febrile seizures (FS) are common in pediatric epilepsy, but their precise etiology remains unclear. Although cytokines such as interleukin (IL)-4, IL-1, and IL-6 are known to influence FS fever responses, their specific role is not fully understood. This study aimed to clarify the correlation between IL-4 levels and febrile convulsions, exploring molecular mechanisms through bioinformatics and animal experiments to enhance the understanding of FS pathogenesis. Material and methods: With the GSE28674 dataset, the K-means clustering algorithm was used to select key genes that regulate IL-4 during feature selection. An animal model of FS was developed, and in vivo experiments were conducted using enzyme-linked immunosorbent assay, flow cytometry, quantitative polymerase chain reaction (qPCR), Western blot, and immunofluorescence for validation. Results: In this study, bioinformatics analysis and K-means clustering identified proto-oncogene (Jun), protooncogene (Fos), and Early growth response-1 (Egr1) as upstream regulators of IL-4. Dual-luciferase reporter assays confirmed that these transcription factors could activate the IL-4 promoter. qPCR and Western blot analyses showed that the messenger RNA (mRNA) and protein expression levels of Jun, Fos, Egr1, and IL-4 in the FS group were significantly higher than those in the normal control (NC) group (P < 0.05). In addition, immunohistochemical analysis demonstrated that the expression levels of these proteins in the FS group were significantly higher than those in the NC group (P < 0.001). The study also explored the impact of the IL-4 receptor blocker dupilumab on FS, revealing that the dupilumab group exhibited significantly reduced seizure latency (P < 0.001) and significantly increased seizure duration and Racine scores versus the FS group (P < 0.01). Furthermore, dupilumab significantly decreased the expression of IL-1β, tumor necrosis factor α, and IL-6 in serum (P < 0.001), as well as heat shock protein 70 mRNA and protein expression, glial fibrillary acidic protein, cysteine protease-3, and Bcl-2-associated X protein/B-cell lymphoma 2 in hippocampal tissue (P < 0.001). Flow cytometry analysis indicated an increased T helper cell type 1 (Th1)/T helper cell type 2 (Th2) cell ratio (P < 0.001) and increased apoptosis in the FS and dupilumab groups versus the NC group (P < 0.001), along with decreased cell cycle progression and proliferation ability (P < 0.001). Compared with the FS group, the dupilumab group exhibited a further increase in Th1/Th2 cell ratio and apoptosis (P < 0.001), along with a further decrease in cell cycle progression and proliferation ability (P < 0.001). Conclusion: IL-4 and its upstream transcription factors Jun, Fos, and Egr1 may be associated with FS occurrence and development. Moreover, dupilumab appeared to mitigate the symptoms of FS and modulate the associated immune response by blocking the IL-4 receptor.