抑制葡糖脑苷脂酶对多巴胺神经细胞内α-突触核蛋白寡聚体形成及细胞自噬功能的影响

doi:10.3969/j.issn.1006-7795.2013.06.008

首都医科大学学报 ›› 2013, Vol. 34 ›› Issue (6): 820-825.doi: 10.3969/j.issn.1006-7795.2013.06.008

• 帕金森病和卒中诊断与发病机制 • 上一篇下一篇

抑制葡糖脑苷脂酶对多巴胺神经细胞内α-突触核蛋白寡聚体形成及细胞自噬功能的影响

刘光伟^1,2, 尹娜^1,2, 弥娜^1,2, 李昕^1,2, 李尧华^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）；国家重点基础发展计划（973）（2011CB504101）；国家科技支撑计划课题（2012BAI10B03）；北京市自然科学基金（7102076）；首都卫生发展科研专项课题（首发2011-1001-01）。

Effect of glucocerebrosidase inhibition on intracellular α-synuclein oligomer formation and autophagic activity of dopaminergic cells

LIU Guangwei^1,2, YIN Na^1,2, MI Na^1,2, LI Xin^1,2, LI Yaohua^1,2, YU Shun^1,2

1. Department of Neurobiology, Xuanwu Hospital, Capital Medical University, Key Laboratory of 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(973 Program) of China(2011CB504101);The National Science and Technology Support Program(2012BAI10B03);Natural Science Foundation of Beijing(7102076);The Capital Health Research and Development Project(SHOUFA 2011-1001-01).

摘要/Abstract

摘要：

目的研究抑制葡糖脑苷脂酶（glucocerebrosidase，GBA）活性对多巴胺神经细胞内α-突触核蛋白（α-synuclein，α-Syn）寡聚体形成及自噬功能的影响。方法在培养MES23.5细胞外添加不同浓度的GBA活性抑制剂（conduritol-β-epoxide，CβE），观察GBA活性的变化，同时在细胞外添加α-Syn单体使其进入细胞内并孵育48 h。使用双抗体夹心ELISA方法测定细胞内α-Syn寡聚体含量，观察细胞自噬活性、氧化应激和凋亡。结果随着CβE质量浓度的增加，GBA活性呈质量浓度依赖性的下降，而细胞内α-Syn寡聚体的含量则呈质量浓度依赖性的增加；在CβE处理细胞，随着细胞外添加的α-Syn单体的质量浓度加大，自噬活性和氧化应激液增强，同时细胞凋亡数量增加。结论抑制GBA活性导致细胞内α-Syn寡聚体增多以及依赖于α-Syn浓度的自噬活性和氧化应激增强和细胞凋亡增加。

关键词: 葡糖脑苷脂酶, α-突触核蛋白寡聚体, MES 23.5细胞, 自噬

Abstract:

Objective To investigate the effect of glucocerebrosidase (GBA) inhibition on intracellular α-synuclein(α-Syn) oligomer formation and autophagic activity of dopaminergic cells. Methods The activity of GBA was measured in MES 23.5 cells treated with different concentrations of CβE, the specific GBA inhibitor. After treatment with CβE, monomeric α-Syn was added into the medium and the amount of intracellular α-Syn oligomers were evaluated by ELISA. The autophagic activity, oxidative stress, and apoptotic cell counts were also examined. Results CβE dose-dependently decreased the activity of GBA and increased the levels of intracellular α-Syn oligomers. CβE also enhanced the autophagic activity, oxidative stress levels and apoptotic cell counts in an α-Syn concentration-dependent manner. Conclusion GBA inhibition causes intracellular accumulation of α-Syn oligomers and α-Syn concentration-dependent enhancement of autophagy, oxidative stress and apoptosis.

Key words: glucocerebrosidase, α-synuclein oligomer, MES 23.5 cells, autophagy

中图分类号:

R338

刘光伟, 尹娜, 弥娜, 李昕, 李尧华, 于顺. 抑制葡糖脑苷脂酶对多巴胺神经细胞内α-突触核蛋白寡聚体形成及细胞自噬功能的影响[J]. 首都医科大学学报, 2013, 34(6): 820-825.

LIU Guangwei, YIN Na, MI Na, LI Xin, LI Yaohua, YU Shun. Effect of glucocerebrosidase inhibition on intracellular α-synuclein oligomer formation and autophagic activity of dopaminergic cells[J]. Journal of Capital Medical University, 2013, 34(6): 820-825.

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抑制葡糖脑苷脂酶对多巴胺神经细胞内α-突触核蛋白寡聚体形成及细胞自噬功能的影响

Effect of glucocerebrosidase inhibition on intracellular α-synuclein oligomer formation and autophagic activity of dopaminergic cells

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