Zheng Fei, Zhou Yan-Tao, Zeng Yi-Fu, Liu Tao, Yang Zhao-Yu, Tang Tao, Luo Jie-Kun, Wang Yang
College of Electrical and Information Engineering, Hunan University, Changsha, China.
Laboratory of Ethnopharmacology, Institute of Integrative Medicine, Xiangya Hospital, Central South University, Changsha, China.
Front Mol Neurosci. 2020 Feb 27;13:27. doi: 10.3389/fnmol.2020.00027. eCollection 2020.
: Stroke is a leading health issue, with high morbidity and mortality rates worldwide. Of all strokes, approximately 80% of cases are ischemic stroke (IS). However, the underlying mechanisms of the occurrence of acute IS remain poorly understood because of heterogeneous and multiple factors. More potential biomarkers are urgently needed to reveal the deeper pathogenesis of IS. : We identified potential biomarkers in rat brain tissues of IS using an iTRAQ labeling approach coupled with LC-MS/MS. Furthermore, bioinformatrics analyses including GO, KEGG, DAVID, and Cytoscape were used to present proteomic profiles and to explore the disease mechanisms. Additionally, Western blotting for target proteins was conducted for further verification. : We identified 4,578 proteins using the iTRAQ-based proteomics method. Of these proteins, 282 differentiated proteins, comprising 73 upregulated and 209 downregulated proteins, were observed. Further bioinformatics analysis suggested that the candidate proteins were mainly involved in energy liberation, intracellular protein transport, and synaptic plasticity regulation during the acute period. KEGG pathway enrichment analysis indicated a series of representative pathological pathways, including energy metabolite, long-term potentiation (LTP), and neurodegenerative disease-related pathways. Moreover, Western blotting confirmed the associated candidate proteins, which refer to oxidative responses and synaptic plasticity. : Our findings highlight the identification of candidate protein biomarkers and provide insight into the biological processes involved in acute IS.
中风是一个主要的健康问题,在全球范围内发病率和死亡率都很高。在所有中风病例中,约80%为缺血性中风(IS)。然而,由于多种异质性因素,急性缺血性中风发生的潜在机制仍知之甚少。迫切需要更多潜在的生物标志物来揭示缺血性中风更深层次的发病机制。
我们采用iTRAQ标记法结合液相色谱-串联质谱(LC-MS/MS)在缺血性中风大鼠脑组织中鉴定潜在的生物标志物。此外,利用基因本体论(GO)、京都基因与基因组百科全书(KEGG)、数据库注释、可视化与综合发现(DAVID)以及Cytoscape等生物信息学分析方法来呈现蛋白质组图谱并探索疾病机制。另外,对目标蛋白进行蛋白质印迹法进一步验证。
我们使用基于iTRAQ的蛋白质组学方法鉴定出4578种蛋白质。在这些蛋白质中,观察到282种差异蛋白,包括73种上调蛋白和209种下调蛋白。进一步的生物信息学分析表明,候选蛋白在急性期主要参与能量释放、细胞内蛋白质运输和突触可塑性调节。KEGG通路富集分析表明存在一系列代表性的病理通路,包括能量代谢物、长时程增强(LTP)以及神经退行性疾病相关通路。此外,蛋白质印迹法证实了与氧化反应和突触可塑性相关的候选蛋白。
我们的研究结果突出了候选蛋白质生物标志物的鉴定,并为急性缺血性中风所涉及的生物学过程提供了见解。