J4 ›› 2012, Vol. 38 ›› Issue (4): 697-700.

• 基础研究 • 上一篇    下一篇

缺血再灌注大鼠脑皮质Bcl-2、Bax和p53的动态表达

许妍妍|刘丽莉*|刘春春*|蔡 睿△|田佳怡#|刘 玲|高振平   

  1. 吉林大学白求恩医学院人体解剖学教研室|吉林 长春 |130021
  • 收稿日期:2012-03-27 出版日期:2012-07-28 发布日期:2012-07-28
  • 通讯作者: 刘 玲(Tel:0431-85619466, E-mail:liuling1978@163.com) E-mail:liuling1978@163.com
  • 作者简介:许妍妍(1982-)|女|吉林省长春市人|在读医学博士|主要从事神经解剖学研究。
  • 基金资助:

    吉林省科技厅科研基金资助课题 (20090185)

Dynamic expressions of Bcl-2,Bax and p53 in cortex of cerebral ischemia-reperfusion rats

XU Yan-yan|LIU Li-li*|LIU Chun-chun*|CAI Rui△| TIAN Jia-yi#,LIU Ling|GAO Zhen-pi     

  1. Department of Human Anatomy|Norman Bethune College of Medicine|Jilin Univer
    sity|Changchun 130021|China
  • Received:2012-03-27 Online:2012-07-28 Published:2012-07-28

摘要:

目的:探讨大鼠脑缺血再灌注损伤早期凋亡相关基因Bcl-2、Bax和p53 mRNA和蛋白表达的动态变化,为研究脑缺血再灌注损伤过程中的神经保护机制提供实验依据。方法:选择健康成年雄性Sprague-Dawley大鼠52只,随机分为假手术对照组(12只)和缺血再灌注2、3、6、8、12 h组(每组8只)。采用插线法制作大鼠大脑中动脉栓塞后再灌注模型;采用原位末端转移酶标记技术(TUNEL)检测脑缺血再灌注不同时间点神经细胞凋亡的变化;通过反转录聚合酶链式反应(RT-PCR)和免疫组织化学法观察凋亡相关基因Bcl-2、Bax和p53表达的动态变化。结果:TUNEL染色显示,缺血再灌注组大鼠脑缺血周围区,再灌注2 h后凋亡细胞开始出现,6~12 h明显增多。Bcl-2、Bax和p53 mRNA和蛋白的表达随着缺血再灌注时间的延长呈增高趋势,Bcl-2蛋白高表达的细胞形态相对完整,Bax和p53蛋白高表达的细胞受损严重。Bcl-2 mRNA和蛋白的表达较Bax和p53出现的早。结论:Bcl-2蛋白的表达对神经细胞具有保护作用,早期调控促进Bcl-2的表达并减少Bax和p53的表达,对降低缺血再灌注过程中神经细胞的损伤有重要作用。

关键词:  脑缺血;再灌注;细胞凋亡

Abstract:

Abstract:Objective To explore the dynamic expressions of apoptosis-related factors of Bcl-2,Bax and p53 in rats with cerebral ischemia-reperfusion injury,and to provide the experimental evidence for the neuroprotective mechanism in cerebral ischemia-reperfusion injury.Methods 52 adult male Sprague-Dawley rats were randomly divided into sham control group (n=12) and cerebral ischemia-reperfusion 2,3,6,8,12 h groups(n=8).Suture method was used to set up the middle cerebral artery occlusion (MCAO) and reperfusion model.The neuronal apoptosis of the MCAO brain tissues at different time points was detected by in situ terminal deoxynucleotidyl transferase labeling technique (TUNEL); the dynamic expressions of Bcl-2,Bax and p53 were detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry.Results The TUNEL staining results  showed that the apoptotic cells began to appear 2 h after reperfusion  in the surrounding area in ischemia-reperfusion group,and they were increased significantly at 6-12 h.The mRNA and protein expressions of Bcl-2,Bax and p53 tended to increase with the prolongaction of ischemia and reperfusion time.The cell morphology was relatively complete in the cells with high expression of Bcl-2 protein,and the cells with high expressions of Bax and p53 protein were severely damaged.The mRNA and protein expressions of Bcl-2 appeared earlier than Bax and p53.Conclusion Bcl-2 has a protective effect on nerve cells.Early regulation can promote the expression of Bcl-2 and reduce the expressions of Bax and p53,and it plays an important role in reducing the damage of nerve cells in the process of ischemia and reperfusion.

Key words: ischemia, reperfusion, apoptosis

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