肢体缺血-再灌注大鼠脑组织iNOS基因表达的半定量检测  

Semiquantitative Detection of iNOS Gene Expression in Brain from Limb Ischemia-Reperfused Rats

作　　者：姚玉霞[1] 史中立[2] 张君岚[2] 丛斌[1] 凌亦凌[2] 

机构地区：[1]河北医科大学实验动物学部,石家庄050017 [2]河北医科大学基础医学院病理生理教研室

出　　处：《中国实验动物学报》2001年第4期251-254,共4页Acta Laboratorium Animalis Scientia Sinica

摘　　要：目的　检测大鼠肢体缺血 再灌注后脑组织iNOS基因表达的变化。方法　SD大鼠 ,随机分为肢体缺血 再灌注 (I R)组、肢体单纯缺血 (I)组及正常对照 (N)组 ,通过夹闭腹主动脉末端 4h ,或 和开放 2～ 2 4h ,复制I、I R组动物模型 ,半定量RT PCR方法检测脑组织iNOSmRNA表达的变化 ,免疫组化染色法观测脑组织内iNOS及过氧亚硝基阴离子 (ONOO- )的硝基化产物硝基酪氨酸 (NT)的生成与分布。结果　N组脑组织iNOSmRNA未检出 ,I组及I R组脑组织iNOSmRNA均有表达 ,再灌注 2h ,iNOSmRNA表达量显著升高 (P <0 .0 1,vsN组 ) ,再灌注 6h达到高峰 ,随后下降 ,2 4h仍有少量表达 ;I及I R组脑组织均有iNOS阳性细胞 ,I R组见于大脑皮层各区、海马、尾状核 ,I组仅见于皮层后肢区及尾状核。I R组脑组织可见弥散分布的NT阳性神经元 ,I组偶见NT阳性神经元。结论　大鼠肢体缺血 再灌后脑组织iNOS基因表达显著增强 。Objective To detect the expression of iNOS gene in brain tissues from limb ischemia\|reperfused SD rats.Method SD rats were randomly divided into three groups:limb ischemia\|reperfusion group(I\|R), simple ischemia group(I) and normal control group(N).The expression of iNOS mRNA in brain was semiquantitatively detected by using of RT\|PCR.The distribution and the expression of iNOS and peroxynitrite protein in brain were measured by immunohistochemistry.Results iNOS mRNA was not detected in brain from control rats,while it was detected in both I and I\|R groups.After I\|R, the expression of iNOS mRNA gradually increased at 2h and peaked at 6h( P <0.01 vs I group),then decreased,but there was also a little expression at 24h.The morphologic results showed that there were more iNOS positive cells and more dispersal NT positive neurons in I\|R group than I group.Conclusion The expression of iNOS mRNA in brain significantly increased after limb ischemia\|reperfusion in a time\|dependent manner.

关 键 词：脑组织 INOS基因 再灌注 肢体缺血 表达 大鼠 尾状核 RNA 阳性神经元 末端 

分 类 号：Q78[生物学—分子生物学] R743[医药卫生—神经病学与精神病学]