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基于串联质谱技术的肌萎缩侧索硬化症患者脑脊液蛋白质组学分析

[Proteomic analysis of the cerebrospinal fluid from patients with amyotrophic lateral sclerosis based on tandem mass spectrometry technique].

作者信息

Su Dandan, Zhang Yong, Bi Fangfang, Xiao Bo

机构信息

Department of Neurology, 928 Hospital of Joint Logistics Support Force of PLA, Haikou 571100, China.

Department of Neurology, Xiangya Hospital, Central South University, Changsha 410008, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2019 Apr 30;39(4):428-436. doi: 10.12122/j.issn.1673-4254.2019.04.08.

Abstract

OBJECTIVE

To investigate the differentially expressed proteins in the cerebrospinal fluid (CSF) of patients with amyotrophic lateral sclerosis (ALS) at the proteomics level using tandem mass spectrometry label (TMT) technique and explore the pathogenic mechanism and related pathways of ALS.

METHODS

Between November, 2017 and April, 2018, 5 patients with medulla oblongata onset ALS and 5 patients with limb onset ALS were selected from the Departments of Neurology of 928 Hospital of Army Joint Logistics Support Force of PLA and Xiangya Hospital of Central South University, with 5 patients with migraine and low intracranial pressure headache serving as the healthy controls.CSF samples were obtained from all the participants, and the differentially expressed proteins in the CSF were identified using tandem mass spectrometry (TMT) technique with bioinformatics analysis.

RESULTS

A total of 1530 proteins were identified and quantified in the CSF samples.The expression of 48 proteins was up-regulated and 6 proteins were down-regulated in medulla oblongata onset ALS patients; 16 proteins were up-regulated and 19 were down-regulated in limb onset ALS patients.GO analysis showed that these proteins, which were distributed both within and outside the cells, were involved in cell physiological process, single organ process and biological regulation and had binding function, catalytic activity, and receptor activity.KEGG pathway analysis showed that the up-regulated proteins in the CSF from patients with medulla oblongata onset ALS participated in 3 pathways involving the lysosomes, metabolism, and measles.The down-regulated proteins in the CSF from patients with limb onset ALS participated in 7 pathways involving the complement and coagulation cascade, infection and herpes simplex infection, and all the pathways contained complement components.

CONCLUSIONS

The CSF samples of ALS patients with medullary onset and limb onset have differentially expressed proteins.The lysosomal pathway is involved in the occurrence and progression of ALS with medullary onset, and the immune responses are involved in the occurrence and progression of ALS with limb onset.

摘要

目的

采用串联质谱标签(TMT)技术在蛋白质组学水平研究肌萎缩侧索硬化症(ALS)患者脑脊液(CSF)中差异表达的蛋白质,探讨ALS的发病机制及相关途径。

方法

2017年11月至2018年4月,从中国人民解放军联勤保障部队第928医院神经内科和中南大学湘雅医院选取5例延髓起病的ALS患者和5例肢体起病的ALS患者,以5例偏头痛和低颅压头痛患者作为健康对照。采集所有参与者的CSF样本,采用串联质谱(TMT)技术结合生物信息学分析鉴定CSF中差异表达的蛋白质。

结果

在CSF样本中总共鉴定并定量了1530种蛋白质。延髓起病的ALS患者中有48种蛋白质表达上调,6种蛋白质表达下调;肢体起病的ALS患者中有16种蛋白质表达上调,19种蛋白质表达下调。基因本体(GO)分析表明,这些蛋白质分布在细胞内和细胞外,参与细胞生理过程、单一器官过程和生物调节,具有结合功能、催化活性和受体活性。京都基因与基因组百科全书(KEGG)通路分析表明,延髓起病的ALS患者CSF中上调的蛋白质参与了3条通路,涉及溶酶体、代谢和麻疹。肢体起病的ALS患者CSF中下调的蛋白质参与了7条通路,涉及补体和凝血级联反应、感染和单纯疱疹感染,所有通路均包含补体成分。

结论

延髓起病和肢体起病的ALS患者CSF样本中有差异表达的蛋白质。溶酶体途径参与延髓起病的ALS的发生和发展,免疫反应参与肢体起病的ALS的发生和发展。

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本文引用的文献

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Evidence for peripheral immune activation in amyotrophic lateral sclerosis.肌萎缩侧索硬化症中外周免疫激活的证据。
J Neurol Sci. 2014 Dec 15;347(1-2):90-5. doi: 10.1016/j.jns.2014.09.025. Epub 2014 Sep 28.

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