Wang Qian, Pan Liangyu, Chen Siruan, Zhang Yuyu, Liu Guangyuan, Wu Yiying, Qin Xia, Zhang Panpan, Zhang Wei, Zhang Jianghua, Kong Dezhi
Institute of Integrative Medicine, College of Chinese Integrative Medicine, Hebei Medical University, Shijiazhuang, China.
Key Laboratory of Tranquilizing TCM, Hebei Provincial Administration of Traditional Chinese Medicine, Shijiazhuang, 050017, China.
Fluids Barriers CNS. 2025 May 13;22(1):49. doi: 10.1186/s12987-025-00660-x.
Complex febrile seizures (CFS) have been associated with an increased risk of epilepsy in adulthood. However, the specific link between blood-brain barrier (BBB) and the predisposition to epilepsy in adults who experienced CFS during infancy remains unclear. The objective of this study was to investigate the alteration of BBB in adult mice who had experienced CFS during infancy, and to explore the mechanisms of increased susceptibility to epilepsy after CFS.
The CFS pup model was induced using hot air, and the seizure susceptibility was examined using low-dose pentylenetetrazole (PTZ) after 8 W. The brain microvessels representing BBB function were isolated and their protein expression changes were analyzed using data-independent acquisition (DIA) proteomic techniques. Subsequently, the bioinformatic analyses were performed using ClusterProfiler, STRING, Gene Set Enrichment Analysis (GSEA), etc. The enriched pathways, changes in the expression of BBB-related proteins, and alterations in metabolites including certain neurotransmitters were subsequently validated by Western Blotting, quantitative real-time polymerase chain reaction (qRT-PCR), and mass spectrometric imaging (MSI). In addition, we selected the MMP inhibitor Incyclinide to verify that dysregulation of the ECM-receptor interaction signaling pathway increases epilepsy susceptibility in adult mice.
Mice that experienced CFS in infancy show increased susceptibility to epilepsy in adulthood, and BBB proteomic profile was significantly altered in the CFS mice. The network analysis suggests that dysregulation of the extracellular matrix (ECM)-receptor interaction pathway is a key mechanism. Moreover, MSI analysis uncovered notable changes in differential metabolites, including amino acids and nucleotide-derived neurotransmitters associated with the function of BBB maintaining neuronal homeostasis. Subsequent validation experiments showed that dysregulation of the ECM-receptor interaction signaling pathway exacerbated epilepsy susceptibility in adult mice.
Our research represents the pioneering demonstration of the modified BBB proteomics associated with epilepsy susceptibility in adult mice previously exposed to CFS in infancy. Notably, the increased susceptibility is attributed to the dysregulation of the ECM-receptor interaction pathway. These findings may help to elucidate the role of BBB alterations in the progression of epilepsy susceptibility, and provide new orientations for subsequent prevention and treatment of epilepsy.
复杂性热性惊厥（CFS）与成年后癫痫风险增加有关。然而，血脑屏障（BBB）与婴儿期经历过CFS的成年人癫痫易感性之间的具体联系仍不清楚。本研究的目的是调查婴儿期经历过CFS的成年小鼠血脑屏障的改变，并探讨CFS后癫痫易感性增加的机制。
使用热空气诱导建立CFS幼鼠模型，并在8周后使用低剂量戊四氮（PTZ）检测癫痫易感性。分离代表血脑屏障功能的脑微血管，使用数据非依赖采集（DIA）蛋白质组学技术分析其蛋白质表达变化。随后，使用ClusterProfiler、STRING、基因集富集分析（GSEA）等进行生物信息学分析。随后通过蛋白质免疫印迹法、定量实时聚合酶链反应（qRT-PCR）和质谱成像（MSI）验证富集途径、血脑屏障相关蛋白表达变化以及包括某些神经递质在内的代谢物变化。此外，我们选择基质金属蛋白酶抑制剂Incyclinide来验证细胞外基质-受体相互作用信号通路的失调会增加成年小鼠的癫痫易感性。
婴儿期经历过CFS的小鼠在成年后癫痫易感性增加，且CFS小鼠的血脑屏障蛋白质组学特征发生了显著改变。网络分析表明，细胞外基质（ECM）-受体相互作用途径的失调是关键机制。此外，MSI分析发现差异代谢物有显著变化，包括与血脑屏障维持神经元稳态功能相关的氨基酸和核苷酸衍生神经递质。随后的验证实验表明，细胞外基质-受体相互作用信号通路的失调加剧了成年小鼠的癫痫易感性。
我们的研究首次证明了与婴儿期曾患CFS的成年小鼠癫痫易感性相关的血脑屏障蛋白质组学改变。值得注意的是，易感性增加归因于细胞外基质-受体相互作用途径的失调。这些发现可能有助于阐明血脑屏障改变在癫痫易感性进展中的作用，并为后续癫痫的预防和治疗提供新方向。
