脑缺氧缺血新生大鼠诱生型一氧化氮合酶基因的表达及其与脑细胞凋亡的关系  

The Relationship between the Gene Expression of Inducible Nitric Oxide Synthase and Brain Cells Apoptosis following Cerebral Hypoxia-ischemia

作　　者：蒲蜀湘[1] 周伟[2] 吴圣楣[3] 陈惠金[3] 

机构地区：[1]广州医学院附属第二医院神经内科,广东广州510260 [2]广州市儿童医院新生儿科,广东广州510120 [3]上海市儿科医学研究所,上海200092

出　　处：《中国神经免疫学和神经病学杂志》2003年第2期123-126,共4页Chinese Journal of Neuroimmunology and Neurology

基　　金：广东省重点科技攻关基金资助项目 (1999-2 45 -3 3 )

摘　　要：目的　探讨新生大鼠缺氧缺血后脑内诱生型一氧化氮合酶 (i NOS)基因表达的变化及其与脑缺氧缺血所致细胞凋亡的关系。方法　建立新生大鼠缺氧缺血性脑损伤动物模型 ,应用快速竞争性逆转录 PCR技术及原位末端标记法观察脑缺氧缺血后不同时间点缺氧缺血侧大脑组织中 i NOS m RNA的表达及神经细胞凋亡的情况。结果　新生大鼠缺氧缺血后 ,缺氧缺血侧大脑 i NOS m RNA表达自 6h开始明显增强 ,其相对表达量为(0 .41± 0 .1 3 ) ;2 4h达高峰 ,相对表达量为 (2 .0 2± 0 .2 4) ,7d时回至基线水平。缺氧缺血侧脑组织中凋亡细胞数目亦自缺氧缺血后 6h开始明显增多 ,平均为 (2 1 .3± 3 .5 ) /1 0个高倍视野 (1 0 hpf) ;2 4h达高峰 ,约 (63 .7±3 .2 ) /1 0 hpf,与对照组 [平均为 (7.3± 2 .3 ) /1 0 hpf]比较差异有显著性 (P <0 .0 1 )。i NOS m RNA的表达高峰与缺氧缺血后脑细胞凋亡的高峰时相相吻合。结论　缺氧缺血可使新生大鼠脑 i NOS m RNA的表达增强和凋亡细胞增加 ,i NOS过表达在缺氧缺血后脑细胞凋亡的调控过程中起一定作用。Objective To investigate the relationship between the gene expression of theinduciblenitric oxidesynthase(iNOS)andbraincellsapoptosisfollowing cerebral hypoxia ischemia in the neonatal rats.Methods 7 day old rat pups were subjected to a unilateral ligation of the common carotid artery followed by a 2 h 30 min stay in the prearranged hypoxic chamber (8%O 2 in N 2). Rapid competitive reverse transcriptase PCR and in situ end labeling methods were used to detect the expression of iNOS mRNA and brain cells apoptosis separately in the hemisphere subjected to both ligation and hypoxia suffered at different time points from hypoxia ischemia.Results In the ipsilateral hemisphere following cerebral hypoxia ischemia, the expression ofiNOSmRNA began at 6 h(0.41±0 13), peaked at 24 h(2 02±0 24), and returned to baseline at 7 days. The evident increase of apoptotic cells in the ipsilateral hemisphere began at 6 h following hypoxia ischemia[(21 3±3 5)/10 hpf], peaked at 24 h[(63 7±3 2)/10 hpf], which was higher than that of the control[(7 3±2 3)/10 hpf, P<0 01]. The peak time of the overexpression of iNOS mRNA was coordinated with that of apoptosis following hypoxia ischemia.Conclusions The increase in expression of iNOS mRNA and the brain cell apoptosis could be induced by cerebral hypoxia ischemia in neonatal rats.The overexpression of iNOS might play a role in the induction of apoptosis following cerebral hypoxia ischemia.

关 键 词：脑缺氧缺血 新生大鼠 诱生型一氧化氮合酶 基因表达 细胞凋亡 迟发性神经元死亡 

分 类 号：R743[医药卫生—神经病学与精神病学]