摘要
目的探讨七氟烷预处理是否减轻脑缺血再灌注引起的神经细胞凋亡。方法36只雄性SD大鼠(250—300g),随机分为5组:对照组(n=12);缺血再灌注组(n=12),动物建立大脑中动脉阻断再灌注模型;七氟烷预处理组(n=12),动物接受七氟烷预处理后建立大脑中动脉阻断再灌注模型。采用HE染色和透射电镜观察大鼠缺血再灌注脑区神经元的凋亡情况、原位细胞凋亡检测法计数神经元凋亡密度、免疫印迹(Westernblot)检测缺血脑区半胱氨酸蛋白酶Caspase-3表达及活化。结果HE染色、电镜的结果均提示七氟烷预处理组神经元凋亡比缺血再灌注组少、凋亡程度轻;神经元凋亡密度对照组为(13.0±1.4)个/0.1mm^2、缺血再灌注组为(189.8±6.8)个/0.1mm^2、七氟烷预处理组为(110.5±4.3)+/0.1mm^2,两两比较,差异有统计学意义;缺血脑区Caspase-3前体及其20000切割片段含量的相对灰度值分别为16.7±3.0、76.1±3.4、51.2±3.1及8.2±2.3、59.0±6.3、31.2±5.4。结论七氟烷预处理可通过减轻脑缺血再灌注引起的神经细胞凋亡而产生保护作用。
Objective To investigate if sevoflurane preconditioning attenuate neuronal apoptosis induced by isehemia-reperfusion. Methods Thirty-six male SD rats weighing 250 - 300 g were randomly divided into three groups (n = 12 each) : control group (group C), isehemia-reperfusion group (group IR) (rats were established cerebral artery clamped and reperfusion model), sevoflurane preconditioning group (group S) (rats were established cerebral artery clamped and reperfusion model after 1 h 2.4% sevoflurane preconditioning). Apoptosis neurons were observed by Hematoxylin and Eosin (HE) staining and transmission electron microscope, TdT mediated Dutp nick end labeling(TUNEL) method was used to count apoptosis neurons density, fresh ischemic brain tissue was taken out, while Caspase-3 zymogen and 20000 segment were checked by Western blot. Results apoptosis neurons in group IR were more than ones in group S under HE staining and light microscope and transmission electron microscope, and apoptosis neurons density (cell number/0. 1 mm^2) by TUNEL staining : group C, 13.0± 1.4; group IR, 189. 8± 6. 8; group S, 110. 5 ±4. 3, the relative gray values of the contents of procaspase-3 and its 20 000 cleavage fragment were 16. 72 ±3. 0, 76. 1 ±3.4, 51.2 ±3.1 and 8. 2 ±2. 3, 59.0 ±6. 3, 31.2±5.4 respectively. Conclusions Sevoflnrane pretreatment can protect neuron on ischemia-reperfusion injury by attenuating neuronal apoptosis in rats.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2009年第41期2943-2945,共3页
National Medical Journal of China
关键词
缺血
再灌注损伤
脑
七氟烷
Ischemia
Reperfusion injury
Brain
Sevoflurane