南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (04): 428-.doi: 10.12122/j.issn.1673-4254.2019.04.08

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肌萎缩侧索硬化患者的脑脊液存在差异表达蛋白：基于串联质谱标签方法

苏丹丹，张勇，毕方方，肖波   

  • 出版日期:2019-04-20 发布日期:2019-04-20

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

  • Online:2019-04-20 Published:2019-04-20

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摘要： 目的基于串联质谱标签（TMT）技术，从蛋白质组学层面研究肌萎缩侧索硬化（ALS）患者脑脊液（CSF）中的差异性表达 蛋白，探讨ALS发病的机制和相关作用途径。方法选取2017年11月~2018年4月于中国人民解放军联勤保障部队第928医院 和中南大学湘雅医院神经内科就诊5例延髓起病ALS患者和5例肢体起病ALS患者，以5例偏头痛及低颅压头痛的患者为健康 对照，分别获取CSF，利用串联质谱标签（TMT）技术鉴定CSF中差异性蛋白质，并进行生物信息学分析。结果共鉴定并定量 1530种蛋白。延髓起病ALS患者中48种蛋白表达上调，6种蛋白表达下调；肢体起病ALS患者中16种蛋白表达上调，19种蛋 白表达下调。GO分析显示这些蛋白均参与细胞生理过程、单器官过程和生物调节等过程，在细胞内和细胞外均有分布，且具有 结合功能、催化活性和受体活性等。KEGG通路分析显示，延髓起病ALS患者CSF中上调蛋白参与溶酶体和代谢等3条通路。 肢体起病ALS患者CSF中下调蛋白参与补体和凝血级联途径、金黄色葡萄球菌感染和单纯疱疹感染等7条通路，各通路均包含 补体成分。结论延髓起病和肢体起病的ALS患者CSF中均存在差异性表达蛋白。溶酶体通路参与延髓起病ALS的发生和发 展，免疫反应参与肢体起病ALS的发生和发展。

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, Staphylococcus aureus infection and herpes simplex infection, and all the pathways contained complement components. Conclusion 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.