大鼠肢体缺血再灌注后海马N-甲基-D-天冬氨酸受体2B亚单位表达的变化  

Changes in protein expression of N-methyl-D-aspartic acid receptor 2B subunit in hippocampus of rats following ischemia-reperfusion of limbs

作　　者：徐殿国[1] 史中立[2] 宋永红[1] 

机构地区：[1]河北工程大学医学院人体解剖系,河北省邯郸市056029 [2]河北医科大学临床学院,河北省石家庄市050017

出　　处：《中国临床康复》2006年第22期110-112,F0003,共4页Chinese Journal of Clinical Rehabilitation

基　　金：河北省自然科学基金资助项目(C2005000729)~~

摘　　要：目的:观察肢体缺血再灌注后海马N-甲基-D-天冬氨酸受体2B亚单位表达的变化。方法:实验于2005-04在河北医科大学解剖系中心实验室完成。选用10～12周龄健康雄性SD大鼠66只,随机数字表法分为3组:正常组(n=6)、假手术模型组、缺血-再灌注组,后2组又分为6,12,24,48及72h5个时间点,每个时间点6只。制备大鼠肢体缺血-再灌注模型,夹闭双侧股动脉根部4h,去夹再分别开放6,12,24,48,72h,假手术模型组同样手术处理,但不夹闭股动脉,不环扎橡皮筋。将正常组、不同时间点(6,12,24,48及72h)的缺血-再灌注组以及假手术模型组大鼠,常规灌注取材及制备石蜡切片,免疫组织化学染色并利用图像分析系统分析。结果:在实验过程中,实验动物无脱失值,全部进入结果分析。①肢体缺血-再灌注6,12,24,48,72h后,N-甲基-D-天冬氨酸受体2B亚单位蛋白在海马CA1区锥体层神经元表达灰度值分别为(14.64±2.71),(18.90±1.40),(24.71±2.89),(22.10±2.62)和(21.62±1.98),与正常组和假手术模型组灰度值比较,缺血-再灌注6h未见显著变化,再灌注12,24,48,72h的表达都有显著升高,差异均有显著性意义(P<0.01/0.05)。其中再灌注24h前后达到峰值,灰度值是正常组的172%。②肢体缺血-再灌注6,12,24,48,72h后,N-甲基-D-天冬氨酸受体2B亚单位蛋白在海马CA3区锥体层神经元表达灰度值分别为(10.47±2.13),(14.26±1.54),(26.32±2.83),(16.18±2.43)和(15.60±2.70),与正常组和假手术模型组灰度值比较,缺血-再灌注6h组未见显著变化,再灌注12,24,48,72h的表达都有显著升高,差异均有显著性意义(P<0.01/0.05);其中再灌注24h前后达到峰值,灰度值是正常组的261%。③假手术模型组与正常组比较,N-甲基-D-天冬氨酸受体2B亚单位蛋白在海马CA1、CA3区锥体层神经元表达灰度值未见显著性变化。结论:N-甲基-D-天冬氨酸1/2B型受体介入了肢体缺血-再灌注致海马损伤的病理过程。AIM： To observe the changes in expression of N-methyl-D-aspartic acid （NMDA） receptor subunit NR2B in hippocampal subfileds of rats following ischemia-reperfusion of limbs. METHODS： The experiment was carried out in the central laboratory of Department of Anatomy, Hebei Medical University in April 2005. Totally 66 healthy male SD rats aged 10-12 weeks were randomly divided into three groups： normal group （n=6）, sham-operation group and ischemia-reperfusion group, and the last two groups were subdivided into reperfusion 6, 12, 24, 48 and 72 hours groups with 6 rats in each time point. After the occlusion of the bilateral arterial root for 4 hours, the clamp was removed, and the models were made after reperfusion for 6, 12, 24, 48 and 72 hours, respectively. The sham-operation group was also given operation but without occlusion of artery. The sections were prepared after routine perfusion sampling of the normal group, ischemia-reperfusion with different time points groups and sham-operation group for analysis by immunohistochemistry-stain method and image analysis system. RESULTS： All the rats were involved in the result analysis, without drop. ①Immunoreactivity gray scales of CA1 subfield in rat stratum pyramidale neuron of hippocampus at 6, 12, 24, 48 and 72 hours after reperfusion were （14.64±2.71）, （18.90±1.40）, （24.71±2.89）, （22.10±2.62） and （21.62 ±1.98）, respectively, which were significantly increased compared with those in the normal group and sham-operation group apart from the 6 hour roperfusion group, and there were significant differences （P 〈 0.01, 0.05）. It reached peak at about 24 hours after reperfusion, and was 172% of that in the normal group. ②Immunoreactivity gray scales of CA3 subfield in rat stratum pyramidale neuron of hippocampus at 6, 12, 24, 48 and 72 hours after reperfusian were （10.47±2.13）, （14.26±1.54）, （26.32±2.83）, （16.18 ±2.43） and （15.60±2.70）, respectively, which were significantly increased compared wit

关 键 词：鼠 海马 缺血 再灌注 氨基酸类 肽类和蛋白质类/D-天冬氨酸 

分 类 号：R338.64[医药卫生—人体生理学]