多巴胺诱导SH-SY5Y细胞帕金森病模型中GRP78的表达

J4 ›› 2010, Vol. 36 ›› Issue (1): 76-77.

多巴胺诱导SH-SY5Y细胞帕金森病模型中GRP78的表达

田明秀^1,2, 常明¹, 朱本清², 解洪荣¹, 王丹平¹, 张春光³, 胡林森¹

（1.吉林大学第一医院神经内科|吉林长春 130021；2.天津市人民医院内科|天津 300130；3.武警吉林省总队医院骨科|吉林长春 130052）

收稿日期:2009-10-08 出版日期:2010-01-28 发布日期:2010-01-28
通讯作者: 胡林森 E-mail:(Tel：0431-85612419,E-mail：hulinsen@hotmail.com)
作者简介:田明秀（1971-）|女|吉林省蛟河市人|主治医师|在读医学博士|主要从事脑血管病以及神经变性疾病的研究。
基金资助:
吉林省科技厅科研基金资助课题（200505200）

(1.Department of Neurology| First Hospital,Jilin University|Changchun 130021|China;2.People’s Hospital of Tianjin,Tianjin 300130,China；3.Deparment of Orthopedics|Armed Police Hospital of Jinlin Province|Changchun 130052|China)

TIAN Meng-Xiu^1,2, CHANG MIng², ZHU Ben-Qing², XIE Hong-Rong¹, WANG Dan-Peng¹, ZHANG Chun-Guang³, HU Lin-Sen¹

(1.Department of Neurology| First Hospital,Jilin University|Changchun 130021|China;2.People’s Hospital of Tianjin,Tianjin 300130,China；3.Deparment of Orthopedics|Armed Police Hospital of Jinlin Province|Changchun 130052|China)

Received:2009-10-08 Online:2010-01-28 Published:2010-01-28

摘要/Abstract

摘要：

目的:探讨神经毒素多巴胺(DA)诱导SH-SY5Y细胞帕金森病(PD)细胞模型中葡萄糖调节蛋白78(GRP78)的表达，为深入研究PD的发病机制提供理论依据。方法：建立多巴胺诱导SH-SY5Y细胞帕金森病模型，实验组加入DA(溶解于0.2%维生素C,DA终浓度为200 μmol•L^-1)，对照组加入等体积的0.2%维生素C,分别孵育24 h，台盼蓝染色检测细胞存活率，Hoechst33342染色检测细胞凋亡。分别提取实验组和对照组细胞总蛋白，应用荧光差异双向凝胶电泳(DIGE)技术获得蛋白点的差异表达，应用基质辅助激光解吸／电离飞行时间质谱仪(MALDI-TOF MS)鉴定差异蛋白质。结果：实验组细胞存活率为53.2%，与对照组细胞存活率（100%）比较显著下降(P<0.05)。荧光染色，实验组可见细胞核呈固缩状或碎裂状，呈典型的凋亡形态学改变；对照组可见细胞胞核着均匀一致蓝色。DIGE分析，实验组表达增高的一个蛋白质点，经质谱分析鉴定确认为GRP78。结论：多巴胺能够诱导SH-SY5Y细胞凋亡，凋亡过程中GRP78表达增高，提示GRP78增高可能与PD的发病机制有关。

关键词: SH-SY5Y细胞；多巴胺； GRP78；荧光差异凝胶电泳

Abstract:

Abstract:Objective
To investigate the expression level of glucose regulating protein 78(GRP78) in SH-SY5Y cells in the Parkinson’s disease(PD) model induced by dopamine(DA)，and provide the theoretical basis for PD．Methods PD models induced by DA in SH-SY5Y cells were established.The cells in experimental group were treated with DA,and the cells in control group were treated with equal volume of 0.2% vitamin C.The cells in two groups were incubated for 24 h, the viability of SH-SY5Y cells was assayed by Trypan blue staining,Hoechst 33342 fluorescence staining was used to estimate the apoptotic percentage．The proteins of SHSY5Y cells treated with or without DA were extracted 24 h after incubation，and then the maps of the proteins were established by 2D-DIGE(differential gel electrophoresis,DIGE).The altered protein spots were identified with MALDI-TOF MS and database searching.Results The survial rate of SH-SY5Y cells in experimental group(53.2%) was lower than that in control group(100%)(P< 0.05)．The Hoechst fluorescence staining result showed that the nuclei of SH-SY5Y cells in exprerimental group appeared karyopyknosis and fragmentation which was consistent with nuclear morphology.A high expression protein spot in experimental group was confirmed as GRP78 by MALDI-TOF MS． Conclusion DA can induce the apoptosis of SH-SY5Y cells,and the expression of GRP78 is increased in the process of apoptosis．The results demonstrate that the increase of GRP78 may be involved in the pathogenesis of PD．

Key words: S-SY5Y cell； dopamine； GRP78； differentialgel eletrophoresis

中图分类号:

R114

田明秀, 常明, 朱本清, 解洪荣, 王丹平, 张春光, 胡林森. 多巴胺诱导SH-SY5Y细胞帕金森病模型中GRP78的表达[J]. J4, 2010, 36(1): 76-77.

TIAN Meng-Xiu, CHANG MIng, Ben-Qing, XIE Hong-Rong, WANG Dan-Peng, ZHANG Chun-Guang, HU Lin-Sen. (1.Department of Neurology| First Hospital,Jilin University|Changchun 130021|China;2.People’s Hospital of Tianjin,Tianjin 300130,China；3.Deparment of Orthopedics|Armed Police Hospital of Jinlin Province|Changchun 130052|China)[J]. J4, 2010, 36(1): 76-77.