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  

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

作　　者：刘天会[1] 陈瑞[2] 

机构地区：[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

出　　处：《首都医科大学学报》2005年第1期59-62,共4页Journal of Capital Medical University

基　　金：北京市教委科技发展计划基金 (0 0KJ10 8)资助项目

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C5 7BL/ 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 2 0min后再通血流 ,建立脑缺血再灌注模型 ,分别于再灌注 6h、12h、2 4h和 4 8h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 3mRNA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 .0 5 ) ;各I/R组与相应时相点HBO组caspase 3mRNA及酶原表达水平相比差异均无统计学意义 (P >0 .0 5 )。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。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.

关 键 词：caspase-3 海马 HBO 表达 高压氧 小鼠 前脑缺血 转录水平 RNA 半胱天冬酶-3 

分 类 号：R743[医药卫生—神经病学与精神病学] R735[医药卫生—临床医学]