脑红蛋白在蛛网膜下腔出血大鼠脑组织中的表达  

Expression of neuroglobin in rat brain tissue after subarachnoid hemorrhage

作　　者：李维德[1,2] 孙青[1,2] 李松[1,2] 杭春华[1,2] 

机构地区：[1]南方医科大学南京临床医学院 [2]南京军区南京总医院神经外科,210002

出　　处：《中国脑血管病杂志》2013年第8期416-420,共5页Chinese Journal of Cerebrovascular Diseases

基　　金：国家自然科学基金资助项目(81171170);南京军区重点课题(10Z024)

摘　　要：目的研究脑红蛋白(Ngb)在大鼠蛛网膜下腔出血(SAH)模型脑组织中的表达。方法将成年雄性Sprague-Dawley大鼠84只,随机分为对照组(12只)和SAH组(72只),SAH组再分为模型建立后3、6、12、24、48及72 h共6个亚组(每组12只)。通过改良后视交叉池注血法建立大鼠SAH模型。应用Western Blot、实时荧光定量逆转录聚合酶链反应(RT-PCR)法和免疫组化法,检测SAH后不同时间点脑组织中Ngb蛋白及其mRNA的表达水平水平变化和分布。结果①Western Blot结果显示,大鼠正常脑组织中Ngb含量较少(0.56±0.14)。SAH后3 h脑组织中Ngb表达蛋白水平开始升高(0.77±0.16),24 h达到高峰(1.27±0.18),与对照组比较,差异有统计学意义,P<0.05;其后逐渐下降。②实时荧光定量RT-PCR结果显示,大鼠脑组织Ngb mRNA从SAH后3 h开始增加,6 h达高峰,随后逐渐下降,至72 h基本下降到对照组水平。其中6 h及12 h组与对照组相比,差异有统计学意义,均P<0.01。③免疫组化染色结果显示,对照组大鼠脑组织中有少量Ngb表达阳性细胞,但多呈弱阳性表达,且在颞叶皮质表达较多,海马区也有少量表达;SAH后Ngb阳性细胞明显增多,并且多数呈强阳性表达,在颞叶皮质增高最明显。结论大鼠SAH后脑组织内Ngb蛋白及mRNA水平表达均上调,提示Ngb可能参与对SAH后早期脑损伤的保护作用。Objective To study the expression of neuroglobin （Ngb） in rat brain tissue after sub- arachnoid hemorrhage （SAH） . Methods A total of 84 male Sprague-Dawley rats were randomly as- signed to a control group （n = 12） and a SAH （n =72） group. The SAH group was redevided into 6 sub- groups： 3, 6, 12, 24, 48, and 72-hour subgroups （n = 12 for each group）. A model of SAH was induced by modified introducing blood to the chiasmatic cistern, and the control group received no treatment. West- ern blot, real-time reverse transcription polymerase chain reaction （RT-PCR） assay and immunohistochem- istry were used to detect the changes and distribution of the expression levels of Ngb and mRNA in brain tissue at different time points after SAH. Results （~）The results of Western blot showed that Ngb content in the rat normal brain tissue was low （0.56 ~0.14）. The level of Ngb protein began to increase at 3 hours after SAH （0.77 -＋0.16）. It reached the peak at 24 hours. Compared to the control group, the difference was statistically significant （ P 〈 0.05 ）. Then it decreased gradually. （~）The real-time quantitative RT-PCR results showed that Ngb mRNA in rat brain tissue began to increase at 3 hours after SAH. It reached the peak at 6 hours, and then decreased gradually. It did not decrease to the level of the control group until 72 hours. Compared to the control group, there were significant differences between the 6-hour subgroup and 12-hour subgroup （all P 〈 0.01 ）. @Immunohistochemical staining results showed that there were a small amount of Ngb positive cells in the rat brain tissue in the control group, hut mostly showed a weakly positive expression, the expression in the temporal cortex was more and the expression in the hippocampal regions was few ; the expression of Ngb positive cells increased significantly after SAH, and most of them showed strongly positive expression, and in the temporal cortex increased most obviously. Conclusion The expression lev- els of Ngb

关 键 词：蛛网膜下腔出血 脑红蛋白 蛋白质印迹法 免疫组织化学 实时荧光定量逆转录聚合酶链反应 大鼠 

分 类 号：R743.35[医药卫生—神经病学与精神病学]