蛋白磷酸酶2A活力增高减轻α-突触核蛋白引起的SK-N-SH细胞凋亡

doi:10.3969/j.issn.1006-7795.2013.06.011

首都医科大学学报 ›› 2013, Vol. 34 ›› Issue (6): 835-839.doi: 10.3969/j.issn.1006-7795.2013.06.011

• 帕金森病和卒中诊断与发病机制 • 上一篇下一篇

蛋白磷酸酶2A活力增高减轻α-突触核蛋白引起的SK-N-SH细胞凋亡

杨巍巍¹, 杨慧², 于顺¹

1. 首都医科大学宣武医院神经生物学研究室, 北京市老年病医疗研究中心分子诊断实验室, 教育部神经变性病重点实验室, 北京 100053;
2. 首都医科大学基础医学院神经生物学系北京脑重大疾病研究院, 北京 100069

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: 杨慧, 于顺 E-mail:huiyang0105@sina.com;yushun103@yahoo.com.cn
基金资助:
国家重点基础研究发展计划（2011CB504102，2012CB722407）；国家自然科学基金（30970940）；北京市自然科学基金重点项目（7131001）。

Activation of protein phosphatase 2A alleviates α-synuclein-induced apoptosis of SK-N-SH cells

YANG Weiwei¹, YANG Hui², YU Shun¹

1. Department of Neurobiology, Xuanwu Hospital, Capital Medical University, Laboratory of Molecular Diagnosis, Beijing Geriatric Medical Research Center, Key Laboratory of Neurodegenerative Disease, Ministry of Education, Beijing 100053, China;
2. Department of Neurobiology, School of Basic Medical Sciences, Beijing Insititute for Brain Disorders, Capital Medical University, Beijing 100069, China

Received:2013-10-18 Online:2013-12-21 Published:2013-12-13
Supported by:
This study was supported by the National Basic Research Program of China(2011CB504102，2012CB722407);National Natural Science Foundation of China(30970940);Key Program of Natural Science Foundation of Beijing(7131001).

摘要/Abstract

摘要：

目的观察蛋白磷酸酶2A（protein phosphatase 2A，PP2A）活力在α-突触核蛋白（α-Syn）引起的细胞凋亡中的作用。方法在SK-N-SH细胞中瞬时转染空载（myc）和α-Syn质粒，免疫荧光化学方法鉴定基因表达情况。Western blotting法和PP2A酶活力检测试剂盒检测PP2A磷酸化水平及酶活力。TUNEL染色和MTT法检测各组细胞死亡情况。结果过表达α-Syn可致磷酸化PP2A（pPP2Ac）水平显著增高及活力降低，同时导致细胞凋亡增多和细胞活力降低。PP2A激动剂C₂-ceramide可显著逆转α-Syn所致细胞凋亡。结论 PP2A活力增高可减轻α-Syn过表达引起的细胞凋亡。

关键词: α-突触核蛋白, 蛋白磷酸酶2A, 帕金森病, 凋亡

Abstract:

Objective To explore the role of protein phosphatase 2A(PP2A) activity in α-Syn-induced cell apoptosis. Methods SK-N-SH cells were transiently transfected with either myc blank vectors or the myc vectors with α-Syn gene. The expression of α-Syn gene were indentified by immunofluorescence. The PP2A phosphorylation levels and activity were investigated via Western blotting and Cell PP2A Colorimetric Assay Kit, respectively. TUNEL staining and MTT assay were used to evaluate the cell death. Results Compared with control group, α-Syn overexpression in SK-N-SH cells led to phosphorylation of PP2A(high pPP2Ac levels) and inhibition of the enzyme activity. α-Syn overexpression also resulted in the enhancement of apoptotic cells and decrease of cell viable cells. C₂-ceramide, the PP2A activator attenuated α-Syn-induced cell death. Conclusion α-Syn enhanced pPP2Ac levels, and inhibited PP2A activity. Activation of endogenous PP2A alleviates α-Syn-induced apoptosis of SK-N-SH cells.

Key words: α-Synuclein(α-Syn), protein phosphatase 2A(PP2A), Parkinson's disease(PD), apoptosis

中图分类号:

R742.5

杨巍巍, 杨慧, 于顺. 蛋白磷酸酶2A活力增高减轻α-突触核蛋白引起的SK-N-SH细胞凋亡[J]. 首都医科大学学报, 2013, 34(6): 835-839.

YANG Weiwei, YANG Hui, YU Shun. Activation of protein phosphatase 2A alleviates α-synuclein-induced apoptosis of SK-N-SH cells[J]. Journal of Capital Medical University, 2013, 34(6): 835-839.

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蛋白磷酸酶2A活力增高减轻α-突触核蛋白引起的SK-N-SH细胞凋亡

Activation of protein phosphatase 2A alleviates α-synuclein-induced apoptosis of SK-N-SH cells

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