一种检测帕金森患者血浆促进α-突触核蛋白寡聚体形成能力的方法

doi:10.3969/j.issn.1006-7795.2013.06.010

首都医科大学学报 ›› 2013, Vol. 34 ›› Issue (6): 830-834.doi: 10.3969/j.issn.1006-7795.2013.06.010

• 帕金森病和卒中诊断与发病机制 • 上一篇下一篇

一种检测帕金森患者血浆促进α-突触核蛋白寡聚体形成能力的方法

李昕^1,2, 杨巍巍^1,2, 李雁^1,2, 于顺^1,2

1. 首都医科大学宣武医院神经生物学研究室, 教育部神经变性病重点实验室, 北京 100053;
2. 北京市老年病医疗研究中心分子诊断实验室, 北京 100053

收稿日期:2013-10-18 出版日期:2013-12-21 发布日期:2013-12-13
通讯作者: 于顺 E-mail:yushun103@163.com
基金资助:
国家自然科学基金（81071014）；国家重点基础研究发展计划（2011CB504101）；首都卫生发展科研专项课题（首发2011-1001-01）；国家科技支撑计划课题（2012BAI10B03）；北京市自然科学基金（7102076）。

A method for evaluation of α-synuclein oligomer formation in plasma of patients with Parkinson’s disease

LI Xin^1,2, YANG Weiwei^1,2, LI Yan^1,2, YU Shun^1,2

1. Department of Neurobiology, Xuanwu Hospital, Capital Medical University, Key Laboratory for Neurodegenerative Diseases, Ministry of Education, Beijing 100053, China;
2. Beijing Geriatric Medical and Clinical Center, Beijing 100053, China

Received:2013-10-18 Online:2013-12-21 Published:2013-12-13
Supported by:
This study was supported by the National Natural Science Foundation of China(81071014);National Basic Research and Development Program of China(2011CB504101);Capital Health Research and Development Project(SHOUFA 2011-1001-01);National Science and Technology Support Program(2012BAI10B03);Natural Science Foundation of Beijing(7102076).

摘要/Abstract

摘要：

目的建立一种检测帕金森病（Parkinson’s disease，PD）患者血浆促进外源性α-突触核蛋白（α-synuclein，α-Syn）寡聚体形成的方法。方法利用本室自制的鼠抗人α-Syn单克隆抗体建立检测α-Syn寡聚体的酶联免疫吸附测定（enzyme linked immunosorbent，ELISA）方法。将不同质量浓度（200、100、50、25、12.5 μmol/L）重组人α-Syn在正常人和PD患者血浆中分别振荡孵育（37℃、280 r/min）24、48、72、96 h。用所建立的ELISA法检测振荡后血浆中的α-Syn寡聚体含量。结果所建立ELISA方法可以特异性检测α-Syn寡聚体，不识别α-Syn单体。重组人α-Syn在质量浓度为100 μmol/L、血浆稀释3倍以下、振荡孵育48 h条件下可以在PD患者血浆中形成较多的寡聚体，并与对照血浆中形成的α-Syn寡聚体含量对比差异最大。结论成功建立了一种检测PD患者血浆促进α-Syn寡聚体形成能力的方法，该方法可以用于诊断PD患者血浆的潜在病理变化。

关键词: 帕金森病, α-突触核蛋白, 血浆, 寡聚体

Abstract:

Objective To establish a method for evaluation of exogenous α-synuclein(α-Syn) oligomer formation in plasma of patients with Parkinson's disease(PD). Methods An ELISA method was developed by using the 3D5 mouse anti-human α-Syn monoclonal antibody. Various concentrations of α-Syn(200, 100, 50, 25, 12.5 μmol/L) was incubated(37℃, 280 r/min) in PD and control sera for 24, 48, 72, and 96 h. The amounts of α-Syn oligomers formed in the sera were measured using the established ELISA method. Results The ELISA method established specifically detected α-Syn oligmers with no reaction to α-Syn monomers. The best differentiation between the α-Syn oligomer formation in PD and control sera was observed when the recombinant α-Syn concentration was 100 μmol/L, with the incubation time being 48 h and the plasma diluted to less than 1/3. Conclusion A method for evaluation of α-Syn oligomer formation in the sera of PD patients and normal controls was established, which can differentiate the potential pathological changes of PD.

Key words: Parkinson’s disease, α-synuclein, plasma, oligomer

中图分类号:

R338

李昕, 杨巍巍, 李雁, 于顺. 一种检测帕金森患者血浆促进α-突触核蛋白寡聚体形成能力的方法[J]. 首都医科大学学报, 2013, 34(6): 830-834.

LI Xin, YANG Weiwei, LI Yan, YU Shun. A method for evaluation of α-synuclein oligomer formation in plasma of patients with Parkinson’s disease[J]. Journal of Capital Medical University, 2013, 34(6): 830-834.

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一种检测帕金森患者血浆促进α-突触核蛋白寡聚体形成能力的方法

A method for evaluation of α-synuclein oligomer formation in plasma of patients with Parkinson’s disease

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