鱼藤酮对MN9D细胞蛋白磷酸酶2A活力的影响

doi:10.3969/j.issn.1006-9771.2018.01.007

《中国康复理论与实践》 ›› 2018, Vol. 24 ›› Issue (1): 37-42.doi: 10.3969/j.issn.1006-9771.2018.01.007

鱼藤酮对MN9D细胞蛋白磷酸酶2A活力的影响

王屹², 杜婷婷³, 刘舒佳², 刘佳⁴, 卢勇泉⁴, 杨慧⁴

1.中国康复研究中心北京博爱医院,a.检验科;b. 脊柱脊髓外科,北京市 100068
2.首都医科大学康复医学院,北京市 100068
3.首都医科大学附属北京天坛医院,北京市神经外科研究所,北京市 100050
4.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帕金森病研究所,教育部神经变性病重点实验室,北京市 100069

收稿日期:2017-05-18 修回日期:2017-10-09 出版日期:2018-01-25 发布日期:2018-01-31
通讯作者: 杨慧,女,博士,教授;E-mail:huiyang@ccmu.edu.cn。
作者简介:王屹(1980-),女,汉族,山西大同市人,博士,主治医师,主要研究方向：神经系统疾病。
基金资助:
1.中国康复研究中心科研专项基金项目(No.2017ZX-26); 2.国家重点研究发展计划项目(No.2016YFC1306002)

Influence of Rotenone on Activity of Protein Phosphates 2A in MN9D Cells

WANG Yi², DU Ting-ting³, LIU Shu-jia², LIU Jia⁴, LU Yong-quan⁴, YANG Hui⁴

1. a. Department of Clinical Laboratory b. Department of Spine and Spinal Cord Surgery, Beijing Bo'ai Hospital, China Rehabilitation Research Center, Beijing 100068, China
2. Capital Medical University School of Rehabilitation Medicine, Beijing 100068, China
3. Beijing Tiantan Hospital, Capital Medical University, Institute of Neurosurgery, Beijing 100050, China
4. Department of Neurobiology, Capital Medical University, Center of Parkinson's Disease, Beijing Institute for Brain Disorders, Key Laboratory for Neurodegenerative Disease of the Ministry of Education, Beijing 100069, China

Received:2017-05-18 Revised:2017-10-09 Published:2018-01-25 Online:2018-01-31
Contact: YANG Hui.E-mail:huiyang@ccmu.edu.cn
Supported by:
China Rehabilitation Research Center Project (No. 2017ZX-26) and National Key R&D Plan of China (No. 2016YFC1306002)

摘要/Abstract

摘要： 目的观察鱼藤酮是否诱导蛋白磷酸酶2A (PP2A)的活力下降,并探讨其机制。方法小鼠中脑多巴胺能细胞系MN9D细胞分为正常组(正常培养)、对照组(培养液中加入与鱼藤酮组等体积的二甲基亚砜)、鱼藤酮组(培养液中加入不同浓度鱼藤酮培养24 h)和鱼藤酮+C2组(5 μmol/L C2-神经酰胺预处理8 h后,50 nmol/L鱼藤酮暴露24 h)。MTT法检测细胞活力,Western blotting分析总PP2A水平及酪氨酸磷酸化水平,比色法检测PP2A活性。结果与对照组相比,鱼藤酮组细胞活力明显下降(P<0.01),PP2A催化亚基酪氨酸磷酸化提高(P<0.01),PP2A活性降低(P<0.05)。鱼藤酮+C2组PP2A酪氨酸磷酸化水平较鱼藤酮组明显降低(P<0.01),PP2A活性提高(P<0.05),细胞活力明显提高(P<0.01)。结论鱼藤酮通过提高PP2A催化亚基307位点酪氨酸磷酸化水平,抑制PP2A活性,并可能参与细胞活力下降。可为发现帕金森病药物作用靶点提供参考。

关键词: 帕金森病, 鱼藤酮, 细胞活力, 蛋白磷酸酶2A

Abstract: Objective To study whether and how rotenone reduces the activity of protein phophatase 2A (PP2A).Methods MN9D cells (mouse midbrain dopaminergic cell line) were divided into normal group (normal cultured), control group (dimethyl sulfoxide of same volume with rotenone group was added in medium), rotenone group (50 nmol/L rotenone was added to the culture medium for 24 hours) and rotenone+C2 group (pretreatment of 5 mol/L C2-ceramide for eight hours, and then exposed to 50 nmol/L rotenone for 24 hours). MTT was used to detect cell viability. Total PP2A levels and tyrosine phosphorylation levels were measured with Western blotting. PP2A activity was detected with colorimetric assay.Results Compared with the control group, the cell viability reduced (P<0.01), phosphorylation of tyrosine 307 of PP2A inceased (P<0.01), and activity of PP2A decreased in the rotenone group (P<0.05). And compared with the rotenone group, the cell viability improved (P<0.01), phosphorylation of tyrosine 307 of PP2A deceased (P<0.01), and activity of PP2A increased (P<0.05) in the rotenone+C2 group.Conclusion Rotenone can inhibit activity of PP2A through increasing phosphorylation of tyrosine 307 at the catalytic subunit of PP2A, which might be involved in reducing cell viability, and implicate a new treatment strategy for Parkinson's disease.

Key words: Parkinson's disease, rotenone, cell viability, protein phosphatase 2A

中图分类号:

R742.5

王屹, 杜婷婷, 刘舒佳, 刘佳, 卢勇泉, 杨慧. 鱼藤酮对MN9D细胞蛋白磷酸酶2A活力的影响[J]. 《中国康复理论与实践》, 2018, 24(1): 37-42.

WANG Yi, DU Ting-ting, LIU Shu-jia, LIU Jia, LU Yong-quan, YANG Hui. Influence of Rotenone on Activity of Protein Phosphates 2A in MN9D Cells[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2018, 24(1): 37-42.

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鱼藤酮对MN9D细胞蛋白磷酸酶2A活力的影响

Influence of Rotenone on Activity of Protein Phosphates 2A in MN9D Cells

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