帕金森病患者血浆增强α-突触核蛋白细胞毒性及其机制研究

doi:10.3969/j.issn.1006-7795.2019.02.017

首都医科大学学报 ›› 2019, Vol. 40 ›› Issue (2): 244-250.doi: 10.3969/j.issn.1006-7795.2019.02.017

帕金森病患者血浆增强α-突触核蛋白细胞毒性及其机制研究

李杰¹, 谢秀娟¹, 宋炜¹, 潘维花¹, 杨巍巍², 于顺²

1. 北京市大兴区中西医结合医院神经内科, 北京 100076;
2. 首都医科大学宣武医院神经生物学研究室, 北京 100053

收稿日期:2018-12-03 出版日期:2019-03-21 发布日期:2019-04-15
通讯作者: 于顺 E-mail:yushun103@163.com
基金资助:
国家自然科学基金（81371200，81071014，81401042），北京市医院管理局"使命"计划专项经费资助（SML20150803），北京市科学技术委员会资助（Z161100005116011，Z171100000117013）。

Blood plasma of patients with Parkinson's disease increases alpha-synuclein neurotoxicity and its mechanisms

Li Jie¹, Xie Xiujuan¹, Song Wei¹, Pan Weihua¹, Yang Weiwei², Yu Shun²

1. Department of Neurology, Beijing Daxing District Hospital of Integrated Chinese and Western Medicine, Beijing 100076, China;
2. Department of Neurobiology, Xuanwu Hospital, Capital Medical University, Beijing 100053, China

Received:2018-12-03 Online:2019-03-21 Published:2019-04-15
Supported by:
This study was supported by National Natural Science Foundation of China (81371200, 81071014, 81401042), Beijing Municipal Administration of Hospitals' Mission Plan (SML20150803), Beijing Municipal Science & Technology Commission (Z161100005116011,Z171100000117013).

摘要/Abstract

摘要： 目的研究帕金森病（Parkinson's disease，PD）患者血浆对α-突触核蛋白（α-synuclein，α-Syn）细胞毒性的影响及其机制。方法将重组人α-Syn加入PD和对照血浆中，采用免疫印迹法及酶联免疫吸附（enzyme linked immunosorbent assay，ELISA）法鉴定α-Syn寡聚体和磷酸化形成情况。在原代神经培养基中加入终浓度为5 μmol/L的α-Syn单体及寡聚体，同时添加30%（体积分数）PD血浆、正常人血浆和胎牛血浆，孵育14 d后，MTT及流式细胞技术观察细胞死亡情况。酶活力试剂盒检测各组原代神经元中蛋白磷酸酶2A（protein phosphatase 2A，PP2A）酶活力。结果 PD患者血浆较正常受试者（normal subject，NS）血浆可促使更多的α-Syn磷酸化和寡聚化。与NS血浆相比，添加到原代神经元培养物中的PD血浆导致更多细胞在细胞外α-Syn存在下死亡，同时伴有PP2A活力降低。结论 PD患者血浆中可能通过降低PP2A酶活力使α-Syn发生磷酸化，导致其聚集并进一步使得原代培养神经元死亡。

关键词: α-突触核蛋白, 血浆, 帕金森病, 原代神经元, 磷酸化

Abstract: Objective To investigate the effect of plasma of Parkinson's disease (PD) on the cytotoxicity of α-synuclein (α-Syn) and its mechanism. Methods Recombinant human α-Syn was added to PD and normal subject plasma. Western blotting and ELISA method were used to identify α-Syn oligomers and phosphorylation. Add α-Syn monomer and oligomer at a final concentration of 5μM to primary neuronal medium with 30% PD plasma, normal subject plasma and fetal bovine plasma, after 14 days of incubation, MTT and flow cytometry were used to observe the cell death. Enzyme activity kit was used to detect the activity of protein phosphatase 2A (PP2A) enzyme in primary neurons of each group. Results Plasma in PD patients induced more α-Syn phosphorylation and oligomerization than normal subjects (NS) plasma. Compared to NS plasma, PD plasma added to primary neuronal cultures resulted in more cells dying in the presence of extracellular α-Syn, accompanied by decreased protein phosphatase 2A activity. Conclusion The plasma of PD patients may phosphorylate α-Syn by decreasing the activity of PP2A enzyme, leading to aggregation and further death of primary cultured neurons.

Key words: α-synuclein, plasma, Parkinson's disease, neurons, phosphorylation

中图分类号:

R742

李杰, 谢秀娟, 宋炜, 潘维花, 杨巍巍, 于顺. 帕金森病患者血浆增强α-突触核蛋白细胞毒性及其机制研究[J]. 首都医科大学学报, 2019, 40(2): 244-250.

Li Jie, Xie Xiujuan, Song Wei, Pan Weihua, Yang Weiwei, Yu Shun. Blood plasma of patients with Parkinson's disease increases alpha-synuclein neurotoxicity and its mechanisms[J]. Journal of Capital Medical University, 2019, 40(2): 244-250.

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帕金森病患者血浆增强α-突触核蛋白细胞毒性及其机制研究

Blood plasma of patients with Parkinson's disease increases alpha-synuclein neurotoxicity and its mechanisms

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