Table_1_Relationship Between the Pyroptosis Pathway and Epilepsy: A Bioinformatic Analysis.xlsx

PurposeThis study aimed to analyse the correlation between the pyroptosis pathway and epilepsy using bioinformatics analysis technology. We analyzed the expression of gasdermin D (GSDMD) and gasdermin E (GSDME), the key molecules of pyroptosis, in kainic acid-induced epileptic mice.MethodsWeighted gene co-expression network analysis (WGCNA) was used to construct a signed co-expression network from expression data to screen gene sets closely related to epilepsy. The correlation between the module and epilepsy was verified through module conservative analysis, gene ontology (GO) annotation analysis, and correlation analysis with known epilepsy genes. We obtained currently recognized pyroptosis-related molecules through literature review, and correlation analysis was used to evaluate their correlation with epilepsy. Differentially expressed gene (DEG) analysis was used to analyse expression changes of pyroptosis-related molecules at the transcriptome level, compared to the sham group. We subsequently established a kainic acid-induced status epilepticus (SE) model in mice and validated the mRNA and protein expression of GSDMD and GSDME, the key molecules of pyroptosis, by quantitative reverse transcription PCR (qRT-PCR) and western blotting (WB).ResultsUsing WGCNA, module conservative analysis, and correlation analysis with known epilepsy genes, we screened out a module (a gene set of interest) closely related to epilepsy that was prominently enriched in immune and inflammatory-related biological processes. Correlation analysis results suggest that pyroptosis-related molecules are closely related to this module, but have no obvious correlation with others. DEG analysis of molecules associated with pyroptosis suggests that most of the pyroptosis-related molecules had significantly increased expression after SE, such as IL1b, Casp1, Casp4, Pycard, Gsdmd, Nlrp3, Aim2, Mefv, Tlr2, Tlr3, and Tlr4. qRT-PCR and WB analysis confirmed that the mRNA and protein levels of GSDMD in the mouse hippocampus were significantly upregulated after SE. The mRNA expression of GSDME was not different between the epilepsy group and sham group. However, the WB results showed that the expression of full-length GSDME was decreased and GSDME-N-terminus were significantly increased after SE.ConclusionsOur study highlights that the pyroptosis pathway may be closely related to epilepsy. GSDMD and GSDME, the key executive molecules of pyroptosis, will help to understand the pathogenesis of epilepsy and aid in discovering new targets for anti-epileptic drug treatments.

本研究旨在运用生物信息学分析技术，探讨焦亡通路与癫痫之间的相关性。研究团队分析了由兴奋性氨基酸诱导的癫痫小鼠中，焦亡关键分子GSDMD（Gasdermin D）和GSDME（Gasdermin E）的表达情况。研究方法包括：利用加权基因共表达网络分析（Weighted Gene Co-expression Network Analysis, WGCNA）构建表达数据的符号共表达网络，筛选与癫痫密切相关基因集；通过模块保守性分析、基因本体（Gene Ontology, GO）注释分析和与已知癫痫基因的相关性分析，验证模块与癫痫之间的关联；通过文献综述获取已知的焦亡相关分子，并利用相关性分析评估其与癫痫的相关性；通过差异表达基因（Differentially Expressed Gene, DEG）分析，对比假手术组，在转录组水平上分析焦亡相关分子的表达变化。随后，研究团队在小鼠中建立了兴奋性氨基酸诱导的癫痫持续状态（Status Epilepticus, SE）模型，并通过定量逆转录PCR（Quantitative Reverse Transcription PCR, qRT-PCR）和蛋白质印迹（Western Blotting, WB）技术验证了焦亡关键分子GSDMD和GSDME的mRNA和蛋白表达水平。研究结果显示：通过WGCNA、模块保守性分析和与已知癫痫基因的相关性分析，筛选出一个与癫痫密切相关且在免疫和炎症相关生物过程中显著富集的模块；相关性分析结果表明，焦亡相关分子与该模块密切相关，但与其他模块无明显关联；对焦亡相关分子的DEG分析表明，大部分焦亡相关分子在SE后表达显著增加，如IL1b、Casp1、Casp4、Pycard、Gsdmd、Nlrp3、Aim2、Mefv、Tlr2、Tlr3和Tlr4；qRT-PCR和WB分析证实，小鼠海马区GSDMD的mRNA和蛋白水平在SE后显著上调；GSDME的mRNA表达在癫痫组和假手术组间无显著差异。然而，WB结果表明，SE后GSDME的全长表达降低，而GSDME-N-terminus的表达显著增加。研究结论：本研究突显了焦亡通路可能与癫痫密切相关。焦亡的关键执行分子GSDMD和GSDME有助于理解癫痫的发病机制，并有助于发现抗癫痫药物治疗的新的治疗靶点。