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[采用iTRAQ-LC-MS/MS技术筛选弥漫性轴索损伤大鼠脑组织差异表达蛋白]

[Screening of Differential Expression Proteins in Rat Brain Tissues after DAI by iTRAQ-LC-MS/MS].

作者信息

Chen Q, Bai J, Zhang W F

机构信息

Forensic Medical Examination Center of Beijing Public Security Bureau, Beijing 100192, China.

出版信息

Fa Yi Xue Za Zhi. 2017 Aug;33(4):348-352. doi: 10.3969/j.issn.1004-5619.2017.04.003. Epub 2017 Aug 25.

DOI:10.3969/j.issn.1004-5619.2017.04.003
PMID:29219262
Abstract

OBJECTIVES

To screen for the differential expression proteins in brain tissues of SD rat after diffuse axonal injury (DAI) by isobaric tag for relative and absolute quantification-liquid chromatograph-mass spectrometer/mass spectrometer (iTRAQ-LC-MS/MS), and to explore potential biomarkers available for the diagnosis of DAI.

METHODS

Animal models of DAI were established with the Marmarou method as reference, and the subjects were divided into blank control group (=4), sham strike group (=4) and fatal strike group (=4), respectively. The proteins in rat brain tissues were detected by iTRAQ-LC-MS/MS, and bioinformatics analysis and verification were performed on the results and screened for the differential expression proteins.

RESULTS

A total of 2 016 proteins were identified and quantified. The bioinformatics analysis revealed that the proteins had wide distribution and function, and participated in different biological processes. There were 16 proteins showed differential expression in fatal strike group, including one up-regulated expression protein and 15 down-regulated expression proteins. The results of iTRAQ-LC-MS/MS were confirmed by Western blotting method.

CONCLUSIONS

Multiple differential expression proteins in rat brain tissues after DAI can be screened by iTRAQ-LC-MS/MS. This not only indicates a research direction for exploring the pathogenesis of DAI, but also provides potential biomarkers available for the diagnosis of DAI.

摘要

目的

采用相对和绝对定量同位素标记液相色谱-质谱联用仪/质谱仪(iTRAQ-LC-MS/MS)筛选弥漫性轴索损伤(DAI)后SD大鼠脑组织中的差异表达蛋白,探索可用于DAI诊断的潜在生物标志物。

方法

参照Marmarou法建立DAI动物模型,将实验对象分别分为空白对照组(n = 4)、假打击组(n = 4)和致死打击组(n = 4)。采用iTRAQ-LC-MS/MS检测大鼠脑组织中的蛋白质,并对结果进行生物信息学分析和验证,筛选差异表达蛋白。

结果

共鉴定和定量了2016种蛋白质。生物信息学分析显示这些蛋白质分布广泛、功能多样,参与不同的生物学过程。致死打击组中有16种蛋白质表现出差异表达,其中1种为上调表达蛋白,15种为下调表达蛋白。iTRAQ-LC-MS/MS结果经蛋白质免疫印迹法验证。

结论

iTRAQ-LC-MS/MS可筛选出DAI后大鼠脑组织中多种差异表达蛋白。这不仅为探索DAI的发病机制指明了研究方向,也为DAI的诊断提供了潜在的生物标志物。

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