摘要
目的 观察浅低温联合艾芬地尔对大鼠全脑缺血再灌注后海马神经元存活和凋亡诱导因子（AIF）易位的影响,探讨两者在脑复苏中联合作用的机制.方法 80只雄性SD大鼠按随机数字表法分组（n=16）：假手术组（Ⅰ）、模型组（Ⅱ）、艾芬地尔组（Ⅲ）、浅低温组（Ⅳ）和艾芬地尔＋浅低温组（Ⅴ）.Ⅰ组不给予电刺激和心肺复苏;Ⅱ～Ⅴ组致颤（4 min）建立全脑缺血再灌注模型;Ⅲ和Ⅴ组于再灌注即刻腹腔注射艾芬地尔,其余组注射等容量双蒸水;Ⅳ和Ⅴ组于再灌注即刻乙醇涂擦体表,10 min内将直肠温度降至（32±1）℃,维持4h,其余组维持直肠温度（37±1）℃.HE染色观察海马CA1区锥体细胞病理改变及计数;免疫荧光法检测海马CA1区AIF易位;免疫蛋白印迹法半定量测定AIF易位.结果 与Ⅰ组（75.0±3.2）个比,Ⅱ、Ⅲ、Ⅳ和Ⅴ组CA1区锥体细胞计数均减少（P＜0.05）;与Ⅱ组（36.0±1.2）个比,Ⅲ组（46.8±1.3）个、Ⅳ组（49.0±2.7）个和Ⅴ组（61.3±2.6）个均增加（P＜0.05）,Ⅴ组增加最明显（P＜0.05）.与Ⅰ组比,免疫荧光法观察Ⅱ、Ⅲ、Ⅳ和Ⅴ组均出现AIF易位.免疫蛋白印迹法结果显示,与Ⅰ组（0.022±0.003）比,其余各组AIF在细胞核的表达均增加（P＜0.05）;与Ⅱ（1.020±0.029）组比,Ⅲ（0.870±0.016）、Ⅳ（0.820±0.050）、Ⅴ（0.550±0.050）组AIF在细胞核的表达均减少（P＜0.05）,Ⅴ组减少最明显（P＜0.05）.结论 浅低温联合艾芬地尔能更有效地减轻全脑缺血再灌注后神经元损伤,其机制可能是减轻AIF易位后启动的促凋亡作用.
Objective To explore the effects of mild hypothermia combined with ifenprodil on the survival of neuronal and translocation of apoptosis inducing factor （AIF） following global cerebral ischemiareperfusion to understand the mechanism of combination in cerebral resuscitation.Methods Eighty male SD rats were randomly divided into 5 groups of sham （Ⅰ）,model （Ⅱ）,ifenprodil （Ⅲ）,mild hypothermia （Ⅳ） and ifenprodil plus mild hypothermia （Ⅴ） （n =16 each）.Group Ⅰ completed all procedures except for ventricular fibrillation （VF） and cardio pulmonary resuscitation （CPR）.For groups Ⅱ and Ⅴ,the model of global cerebral ischemia-reperfusion was established and VF induced with transoesophageal cardiac pacing; groups Ⅲ and Ⅴ received by an intraperitoneal injection of ifenprodil immediately after reperfusion and other groups had an equal volume of distilled water.Rectal temperature was cooled down to （32 ± 1） ℃ in groups Ⅳ and Ⅴ by rubbing body surface with ethanol in 10 min after reperfusion and maintained 4 hours continuously while other groups at （37 ± 1）℃.In hippocampal CA1 region at 24 hours after reperfusion,the pathomorphological changes and quantity of pyramidal cells were detected with hematoxylin and eosin staining,nuclear translocation of AIF was shown with immunofluorescence technique and the nuclear expression level of AIF was measured with Western blot.Results Compared with group] （75.0 ± 3.2）,the number of pyramidal cells decreased in other groups （P 〈 0.05） ; compared with group Ⅱ （36.0 ± 1.2）,the number increased in group Ⅲ （46.8 ± 1.3）,Ⅳ （49.0 ±2.7）and Ⅴ （61.3 ±2.60） （P 〈0.05）.In particular,cell count increased significantly in group Ⅴ （P 〈 0.05）.Compared to group Ⅰ,the transloctation of AIF form mitochondria to nucleus was detected in other groups ; compared with group Ⅰ （0.022 ± 0.003）,the expression level of AIF in the nucleus was higher in other groups （P 〈 0.05）.Compa
出处
《中华医学杂志》
CAS
CSCD
北大核心
2014年第17期1353-1356,共4页
National Medical Journal of China
关键词
低温
脑缺血
再灌注损伤
凋亡诱导因子
Hypothermia
Brain ischemia
Reperfusion injury
Apoptosis inducing factor
