Abnormal mitochondrial dynamics can lead to seizures, and improved mitochondrial dynamics can alleviate seizures. Vacuolar protein sorting 13D (VPS13D) is closely associated with regulating mitochondrial homeostasis and autophagy. However, further investigation is required to determine whether VPS13D affects seizures by influencing mitochondrial dynamics and autophagy. We aimed to investigate the influence of VPS13D on behavior in a rat model of acute epileptic seizures. Hence, we established an acute epileptic seizure rat model and employed the CRISPR/CAS9 technology to construct a lentivirus to silence the gene. Furthermore, we used the HT22 mouse hippocampal neuron cell line to establish a stable strain with suppressed expression of . Then, we performed quantitative proteomic and bioinformatics analyses to confirm the mechanism by which VPS13D influences mitochondrial dynamics and autophagy, both and using the experimental acute epileptic seizure model. We found that knockdown of resulted in reduced seizure latency and increased seizure frequency in the experimental rats. Immunofluorescence staining and western blot analysis revealed a significant increase in mitochondrial dynamin-related protein 1 expression following knockdown. Moreover, we observed a significant reduction in LC3II protein expression levels and the LC3II/LC3I ratio (indicators for autophagy) accompanied by a significant increase in P62 expression (an autophagy adaptor protein). The proteomic analysis confirmed the up-regulation of P62 protein expression. Therefore, we propose that VPS13D plays a role in modulating seizures by influencing mitochondrial dynamics and autophagy.

中文翻译：

---

VPS13D 通过调节癫痫大鼠线粒体裂变和自噬影响癫痫发作

线粒体动力学异常可导致癫痫发作，而改善线粒体动力学可缓解癫痫发作。液泡蛋白分选 13D (VPS13D) 与调节线粒体稳态和自噬密切相关。然而，需要进一步研究以确定 VPS13D 是否通过影响线粒体动力学和自噬来影响癫痫发作。我们的目的是研究 VPS13D 对急性癫痫发作大鼠模型行为的影响。因此，我们建立了急性癫痫发作大鼠模型，并利用CRISPR/CAS9技术构建了慢病毒来沉默该基因。此外，我们使用HT22小鼠海马神经元细胞系建立了抑制表达的稳定株。然后，我们使用实验性急性癫痫发作模型进行定量蛋白质组学和生物信息学分析，以确认 VPS13D 影响线粒体动力学和自噬的机制。我们发现敲除 导致实验大鼠癫痫发作潜伏期缩短和癫痫发作频率增加。免疫荧光染色和蛋白质印迹分析显示，敲低后线粒体动力相关蛋白 1 的表达显着增加。此外，我们观察到 LC3II 蛋白表达水平和 LC3II/LC3I 比率（自噬指标）显着降低，同时 P62 表达（自噬衔接蛋白）显着增加。蛋白质组分析证实P62蛋白表达上调。因此，我们认为 VPS13D 通过影响线粒体动力学和自噬来调节癫痫发作。