体外模拟脑缺血再灌注OGD/R模型的差异凝胶电泳分析

J4 ›› 2012, Vol. 38 ›› Issue (5): 861-865.

体外模拟脑缺血再灌注OGD/R模型的差异凝胶电泳分析

刘守跃¹|张影²|胡林森³|宋彬¹|罗毅男⁴|关文明¹

（1．吉林大学第二医院神经外科|吉林长春130041；2．吉林大学第一医院儿科|吉林长春 130021；3. 吉林大学第一医院神经内科|吉林长春130021；4. 吉林大学第一医院神经外科|吉林长春130021）

收稿日期:2012-05-03 出版日期:2012-09-28 发布日期:2012-09-28
通讯作者: 关文明 E-mail:（Tel: 0431-88796286,E-mail:lsy2007.2008@yahoo.com.cn）
作者简介:刘守跃（1971-）| 男|吉林省长春市人|主治医师|医学博士|主要从事脑血管病基础与临床研究。 
基金资助:
吉林省科技厅科研基金资助课题（200905151）

Difference gel electrophoretic analysis of OGD/R model in vitro tomimic ischemia and reperfusion

LIU Shou-yue¹,ZHANG Ying²,HU Lin-sen³,SONG Bin1,LUO Yi-nan⁴,GUANG Wen-ming¹

(1.Department of Neurosurgery,Second Hospital,Jilin University,Changchun 130041,China|2. Department of Pediatrics,First Hospital,Jilin University,Changchun 130021,China|3.Department of Neurology,First Hospital,Jilin University,Changchun 130021,China|4.Department of Neurosurgery,First Hospital,Jilin University,Changchun 130021,China)

Received:2012-05-03 Online:2012-09-28 Published:2012-09-28

摘要/Abstract

摘要：

目的：通过体外模拟缺血-再灌注损伤（I/R）诱导细胞凋亡的氧糖剥离再灌注（OGD/R）模型的建立，应用蛋白质组学技术观察凋亡细胞蛋白质组的差异表达，进一步揭示神经元凋亡的发生机制。方法：选用人神经母细胞瘤系SH-SY5Y细胞分为对照组和实验组，实验组给予OGD10h/R24h处理建立OGD/R诱导SH-SY5Y细胞凋亡模型，提取对照组和实验组细胞总蛋白，利用荧光染色双向差异凝胶电泳技术(2D-DIGE) 检测细胞蛋白质组的差异表达变化。结果：DeCyder软件分析2D-DIGE凝胶图像显示平均分离了（1 800±156）个蛋白质点；与对照组比较，实验组共有30个蛋白点的表达水平有显著变化（蛋白表达量上调或下调30％以上），其中22个升高，8个降低，与同时间对照组比较差异具有统计学意义(P<0.05或P<0.01) 。结论：应用双向差异凝胶电泳的蛋白质组学技术在OGD/R模型中发现有差异蛋白的表达。

关键词: 氧糖剥离/再灌注；蛋白质组学；二维荧光差异凝胶电泳

Abstract:

Abstract:Objective
To build oxygen glucose deprivation/reperfusion(OGD/R) model in vitro to mimic ischemia and reperfusion inducing apoptosis of SH-SY5Y cells,to detect differential proteins by employing proteomics technique and to reveal the potential mechanisms of neuronal apoptosis.Methods SH-SY5Y cells were employed and divided into control group and experimental group in which the cells treated with OGD10h/R24h to set up apoptosis model.After the proteins were extracted from SH-SY5Y cells in control and OGD10h/R24h groups,2D-DIGE techniques was used to detect differential proteins.Results By DeCyder software mean (1 800±156) protein spots were displayed on 2D-DIGE image.In OGD10h/R24h group,30 protein spots were detected to differentially express compared with control group,of which 22 spots were up-regulated and 8 spots were down-regulated.Compared with control groups at the same time, the differences were significant (P<0.05 or P<0.01).Conclusion By employing 2D-DIGE proteomics techniques to analyse OGD/R model,the protein spots are detected to differentially express.

Key words: oxygen glucose deprivation/reperfusion, proteomics, 2-dimentional difference gel electrophoresis

中图分类号:

刘守跃|张影|胡林森|宋彬|罗毅男|关文明. 体外模拟脑缺血再灌注OGD/R模型的差异凝胶电泳分析[J]. J4, 2012, 38(5): 861-865.

LIU Shou-yue,ZHANG Ying,HU Lin-sen,SONG Bin,LUO Yi-nan,GUANG Wen-ming. Difference gel electrophoretic analysis of OGD/R model in vitro tomimic ischemia and reperfusion[J]. J4, 2012, 38(5): 861-865.