Department of Neurology, Xiangya Hospital, Central South University, Changsha, P.R. China.
Department of Neurology, Hunan Provincial People's Hospital, Changsha, P.R. China.
Int J Dev Neurosci. 2021 Apr;81(2):125-141. doi: 10.1002/jdn.10082. Epub 2021 Jan 4.
Epilepsy can severely affect the quality of life of patients, who are often at higher risk of mortality. However, the molecular mechanisms and pathogenesis underlying epileptogenesis are poorly understood. In this study, we performed a proteomic analysis of the hippocampus in pentylenetetrazole (PTZ)-kindled epileptic rats to explore the molecular mechanisms of epileptogenesis. We established an epileptic model in Sprague Dawley rats by injecting PTZ intraperitoneally and applied isobaric tags for relative and absolute quantification (iTRAQ) technology integrated with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify differentially expressed proteins (DEPs) in the hippocampus. A total of 99 proteins, comprising 93 upregulated and 6 downregulated proteins, were identified based on a fold change >1.2 (or <0.83) and a p-value < .05. A further bioinformatics analysis suggested that the candidate proteins were mainly involved in the ubiquitin ligase complex or metabolite homeostasis or acted as intrinsic components of the membrane. A Kyoto Encyclopedia of Gene and Genomes (KEGG) pathway enrichment analysis identified a series of representative pathological pathways, including the calcium signaling pathway, neuroactive ligand-receptor interaction pathway, and the NF-kappa B pathway. The mass spectrometry results were further confirmed by assessing five representative proteins (Akt1, Syvn1, Amfr, Lamb1, and Cox17) using western blotting and immunohistochemistry. These results may help to reveal the molecular mechanisms underlying epileptogenesis and provide new directions or targets for epilepsy research.
癫痫会严重影响患者的生活质量，他们往往有更高的死亡率风险。然而，癫痫发生的分子机制和发病机制还了解甚少。在这项研究中，我们对戊四氮（PTZ）点燃的癫痫大鼠海马进行了蛋白质组学分析，以探讨癫痫发生的分子机制。我们通过腹腔注射 PTZ 在 Sprague Dawley 大鼠中建立了癫痫模型，并应用同位素质谱标签相对和绝对定量（iTRAQ）技术与液相色谱-串联质谱（LC-MS/MS）相结合，鉴定海马中的差异表达蛋白（DEPs）。基于 fold change >1.2（或 <0.83）和 p 值 <0.05，共鉴定出 99 种蛋白质，其中包括 93 种上调蛋白和 6 种下调蛋白。进一步的生物信息学分析表明，候选蛋白主要参与泛素连接酶复合物或代谢物稳态，或作为膜的固有成分。京都基因与基因组百科全书（KEGG）通路富集分析鉴定了一系列代表性的病理途径，包括钙信号通路、神经活性配体-受体相互作用途径和 NF-kappa B 途径。通过使用 Western blot 和免疫组织化学评估 5 种代表性蛋白质（Akt1、Syvn1、Amfr、Lamb1 和 Cox17），进一步验证了质谱结果。这些结果可能有助于揭示癫痫发生的分子机制，并为癫痫研究提供新的方向或靶标。
