永久性大脑中动脉闭塞大鼠梗死周围组织p-Akt(Ser473)、p-Bad(Ser136)表达与细胞凋亡  

Expressions of p-Akt （Ser473） and p-Bad （Ser136） and apoptosis in pefiinfarction tissue foUowing pertinent middle cerebral artery occlusion in rats

作　　者：何敏 韩江全[2] 倪白云 张险平 

机构地区：[1]芜湖市第一人民医院神经内科,241000 [2]遵义医学院第五(珠海)附属医院神经内科,珠海519100

出　　处：《国际脑血管病杂志》2013年第8期624-629,共6页International Journal of Cerebrovascular Diseases

基　　金：珠海市重点学科项目（200880）

摘　　要：目的探讨永久性大脑中动脉闭塞（middle cerebral artery occlusion，MCAO）大鼠脑梗死周围组织P—Akt（Ser473）、p-Bad（Serl36）表达的变化及其与细胞凋亡关系。方法60只雄性Sprague-Dawley大鼠随机分为假手术、MCAO3h、MCAO12h、LY294002干预MCAO3h和LY294002干预MCAO12h组，每组12只。改良线栓法制作永久性MCAO模型，LY294002干预组在模型制作前15n血经侧脑室给药。采用Longa法进行神经功能评分，氯化三苯基四氮唑（2,3，5-triphenyl tetrazolium chloride，TTC）法检测梗死体积，免疫组织化学染色法检测梗死周围组织p-Akt（Ser473）和p-Bad（Serl36）表达，TUNEL法检测梗死周围组织凋亡细胞。结果模型制作后3h，所有实验大鼠均从麻醉中清醒。假手术组神经功能缺损评分为0分，MCAO3h组、MCAO12h组、LY294002干预MCAO3h组和LY294002干预MCAO12h组分别为（2．25±0．45）、（2．92±0．99）、（3．00±0．95）和（3．02±0．36）分，各组间存在显著性差异（F=26．520，P=0．000）。其中，LY294002干预MCAO3h组显著性高于MCAO3h组（P=0．009），LY294002干预MCAO12h组与MCAO12h组无显著性差异（P=0．354）。TTC染色显示，假手术组未见梗死灶，MCAO3h组、MCAO12h组、LY294002干预MCAO3h组和LY294002干预MCAO12h组梗死体积分别为（23．4±1．4）、（40．3±1．1）、（31．9±6．0）和（44．4±3．8）mm2，各组间存在显著性差异（F=30．440，P=0．000）。其中，LY294002干预MCAO3h组显著性大于MCAO3h组（P=0．002），LY294002干预MCAO12h组与MCAO12h组无显著性差异（P=0．113）。与假手术组相比，MCAO3h组梗死周围组织p-Akt（Ser473）表达显著性增高，MCAO12h组显著性下降；p-Bad（Ser136）表达水平随着MCAO时间的推移呈现进行性下降，同时凋亡细胞数量进行性增多。LY294002干预后，MCAO3h时梗死周围组织p-Akt（Ser473）和p-Bad（Ser136）表达水平均显著性下降，凋亡细胞数�Objective To investigate the relationship between the expression changes of p-Akt （Ser473）, p-Bad （Ser136） and the cell apoptosis in peri-infarction tissue of permanent middle cerebral artery occlusion （MCAO） in rats. Methods Sixty male Spmgue-Dawley rats were randomly allocated into sham operation, MCAO 3 h, MCAO 12 h, LY294002 intervention MCAO 3 h, and LY294002 intervention MCAO 12 h groups （n = 12 in each group）. A permanent MCAO model was induced by the modified suture method. At 15 minutes before modeling, the rats in the LY294002 intervention MCAO groups were administered via lateral ventricle. The neurological function score was scored by using Zea Longa method. 2,3,5-triphenyltetrazolium chloride （TTC） staining was used to detect infarct volume. Immunohistochemical staining was use to detect p- Akt （Ser473） and p-Bad （Ser136） expressions in peri-infarction tissue. TUNEL assay was used to detect apoptotic cells in peri-infarction tissue. Results Three hours after modeling, all the experimental rats awoke from anesthesia. The neurological deficit score in the sham operation goup was 0, and the scores of the MCAO 3 h, MCAO 12 h, LY294002 intervention MCAO 3 h and LY294002 intervention MCAO 12 h groups were 2. 25 ± 0. 45, 2. 92 ± 0. 99, 3.00 ± 0. 95, and 3.02 ± 0. 36, respectively. There were significant differences among all groups （F = 26. 520, P = 0. 000）. The score of the LY294002 intervention MCAO 3 h group was significantly higher than that of the MCAO 3 h group （P =0. 009）. There was no significant difference between the LY294002 intervention MCAO 12 h group and the MCAO 12 h group （P =0. 354）. TTC staining showed that no infarct was observed in the sham operation group. The infarct volumes of the MCAO 3 h, MCAO 12 h, LY294002 intervention MCAO 3 h and LY294002 intervention MCAO 12 h groups were 23.4 ± 1.4, 40. 3 ± 1.1, 31.9 ± 6. 0 and 44. 4 ± 3.8 mm3, respectively. There were significant differences among the groups （F =30. 440, P =0. 000）. The scor

关 键 词：脑缺血 蛋白质丝氨酸苏氨酸激酶 磷脂酰肌醇3-激酶类 bcl-死亡相关蛋白 原癌基因蛋白质c akt 细胞凋亡 疾病模型 动物 大鼠 

分 类 号：R743.33[医药卫生—神经病学与精神病学]