基于 iTRAQ 和 PRM 的癫痫大鼠海马比较蛋白质组学分析。

iTRAQ and PRM-Based Comparative Proteomic Profiling of the Hippocampus in Rat Models of Epilepsy.

机构信息

Department of Forensic Medicine, Chongqing Medical University, 400010 Chongqing, China.

Faculty of Medical Technology, Chongqing Medical and Pharmaceutical College, 401331 Chongqing, China.

出版信息

J Integr Neurosci. 2023 Jan 16;22(1):21. doi: 10.31083/j.jin2201021.

DOI:10.31083/j.jin2201021

PMID:36722232

Abstract

BACKGROUND

Epilepsy is a disease caused by paroxysmal abnormal supersynchronous electrical activity of brain neurons, and it is also one of the most common illnesses in neurology. Among the causes, hippocampal sclerosis may be one of the main causes of temporal lobe epilepsy. However, the pathogenesis of hippocampal sclerosis in epilepsy remains unclear.

METHODS

We established an epilepsy model by intraperitoneal injection of pentetrazol (PTZ) into Sprague-Dawley rats, and applied isobaric tags for relative and absolute quantitation (iTRAQ) technology to identify differentially expressed proteins (DEPs) in the hippocampus. We quantified a total of 3782 proteins. DEPs were defined as proteins with a fold change >1.2 (or <0.83) and a Q value (-adjusted) <0.05.

RESULTS

Comparing the epilepsy group and the control group, we identified 170 DEPs, comprising 109 upregulated and 61 downregulated proteins. According to bioinformatics analysis, the DEPs were primarily involved in long-term potentiation, the calcium signalling pathway, aldosterone synthesis and secretion, carbon metabolism, and dopaminergic synapses. Four of these proteins were validated using parallel reaction monitoring (PRM), including Glud1, Atp1a2, Prkcg and Arpc3.

CONCLUSIONS

Our research results may provide further insight into the molecular pathology of hippocampal injury in epilepsy.

摘要

背景

癫痫是一种由脑神经元阵发性异常超同步电活动引起的疾病，也是神经病学中最常见的疾病之一。在病因中，海马硬化可能是颞叶癫痫的主要原因之一。然而，癫痫中海马硬化的发病机制尚不清楚。

方法

我们通过腹腔注射戊四氮（PTZ）建立癫痫大鼠模型，应用同位素标记相对和绝对定量（iTRAQ）技术鉴定海马中差异表达蛋白（DEPs）。我们共定量了 3782 种蛋白质。差异表达蛋白（DEPs）定义为倍数变化>1.2（或<0.83）且 Q 值（-调整）<0.05 的蛋白。

结果

与对照组相比，癫痫组共鉴定出 170 个 DEPs，包括 109 个上调蛋白和 61 个下调蛋白。根据生物信息学分析，这些 DEPs 主要参与长时程增强、钙信号通路、醛固酮合成和分泌、碳代谢和多巴胺能突触。使用平行反应监测（PRM）验证了其中的 4 种蛋白质，包括 Glud1、Atp1a2、Prkcg 和 Arpc3。