首都医科大学学报 ›› 2005, Vol. 26 ›› Issue (1): 59-62.

• 论著·基础研究 • 上一篇    下一篇

高压氧对短暂前脑缺血小鼠海马半胱天冬酶-3表达的影响

刘天会1, 陈瑞2   

  1. 1. 首都医科大学附属北京友谊医院肝病中心;2. 首都医科大学生物化学与分子生物学系
  • 收稿日期:2004-07-05 修回日期:1900-01-01 出版日期:2005-02-24 发布日期:2005-02-24

Hyperbaric Oxygen on Expression of Caspase-3 in Mouse Hippocampus after Transient Forebrain Ischemia

Liu Tianhui1, Chen Rui2   

  1. 1. Department of Liver Center, Beijing Friendship Hospital, Affiliate of Capital University of Medical Sciences;2. Department of Biochemistry and Molecular Biology, Capital University of Medical Sciences
  • Received:2004-07-05 Revised:1900-01-01 Online:2005-02-24 Published:2005-02-24

摘要:

旨在通过测定缺血再灌注时半胱天冬酶-3(caspase-3)的表达以及高压氧对表达的影响, 从细胞凋亡的角度探讨高压氧治疗脑缺血的分子生物学机制。将C57BL/6N小鼠分为假手术组(正常对照组)、缺血再灌注组(I/R组)及缺血再灌注加高压氧治疗组(HBO组), 后2组夹闭双侧颈总动脉20min后再通血流, 建立脑缺血再灌注模型, 分别于再灌注6h、12h、24h和48h取海马。采用半定量反转录-PCR及蛋白免疫印迹(WesternBlotting)方法分别检测caspase-3在转录水平(mRNA)及翻译水平的表达变化。结果:各I/R组及HBO组海马中caspase-3 mRNA及酶原表达水平与假手术组相比均有明显升高, 且差异有统计学意义(P<0.05);各I/R组与相应时相点HBO组caspase-3 mRNA及酶原表达水平相比差异均无统计学意义(P>0.05)。提示:脑缺血再灌注损伤诱发caspase-3转录水平增加, 而且这是引起其酶原增加的主要原因;caspase-3介导的细胞凋亡参与了缺血再灌注损伤;本实验中所采用的HBO治疗方案对caspase-3转录及酶原水平无明显作用。

关键词: 脑缺血再灌注, 高压氧, 半胱天冬酶-3, 表达

Abstract:

This study was to observe the effect of hyperbaric oxygen (HBO) on the expressio ns of caspase-3 in hippocampus following transient forebrain ischemia, and to find out the possible mechanism of HBO treatment. C57BL/6N mice were divided into three groups: sham-operated group (control), ischemia/reperfusion(I/R) group and HBO treatmen t after I/R group. Transient forebrain ischemia was induced by bilateral common carotid artery occlusion for 20 min. Hippocampus was obtained at reperfusion ti me poin ts of 6 h, 12 h, 24 h and 48 h respectively after 20 min of ischemia. RT-PCR wa s used to detect the expression of caspase-3 mRNA, and Western Blotting was use d to detect the level of caspase-3 protein. Experimental data showed that caspase-3 mRNA and procaspase-3 level increased in hippocampus in I/R groups and HBO groups. The difference was significant(P< 0.05) when compared with express ion levels in sham-operated mice. There was no significant alteration in expres s ion levels between every I/R group and HBO treatment group of corresponding time point(P> 0.05). These data indicate that ischemia/reperfusion which induces caspase-3 mRNA expression and increases transcription of caspase-3 is the mean reason fo r procaspase-3 increase. Caspase-3-dependent apoptosis is involved in ischemia /re perfusion injury. The HBO treatment project has no significant effect on caspase-3 mRNA and procaspase-3.

Key words: cerebral ischemia/reperfusion, hype rbaric oxygen, caspase-3, expression

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