Gαi2对脑缺血再灌注后神经细胞凋亡影响.docVIP

Gαi2对脑缺血再灌注后神经细胞凋亡影响 【摘要】目的 探讨分析抑制性G蛋白α2亚单位（Gαi2）在大鼠脑缺血-再灌注损伤模型海马中的表达及其对神经细胞内钙离子浓度和凋亡的影响。方法 将90只SD大鼠随机（随机数字法）分为假手术组（SS组，n=30）、脑缺血-再灌注损伤组（IR组，n=30）。脑缺血-再灌注且脑室内微量灌注Gαi2活性抑制剂百日咳毒素（PT）组（PT组，n=30）；用免疫组化、Western blotting分别测定大鼠脑缺血-再灌注后6 h、12 h、24 h各组中海马神经内Gαi2的表达，采用流式细胞术测定各组中海马细胞内游离Ca2+浓度的平均荧光值，TUNEL法检测神经元细胞的凋亡。结果 不同时间点，IR组Gαi2含量和Ca2+浓度较SS组明显增高（P2+浓度较IR组升高（P【关键词】抑制性G蛋白α2亚单位；脑缺血-再灌注损伤；海马；百日咳毒素；凋亡；神经细胞；钙离子；死亡The effect of Gαi2 on neuronal apoptosis in cerebral ischemia-reperfusion injury model of rats LI Jian， LIU Peng， LIAO Yi-liu.Department of Traumatic Surgery， Tongji Hospital， Tongji Medical College of Huazhong University of Science and Technology， Wuhan 430030，ChinaCorresponding author：LIAO Yi-liu， Email： yiliu.liao@【Abstract】Objective To discuss the levels of inhibitory protein Ga-2（Gαi2） in hippocampus of brain and the effects of Gαi2 on neuronal intracellular Ca2+ level in cerebral ischemia reperfusion injury model of rats. Methods Ninety SD rats were randomly（random number） assigned to sham group （n=30）， ischemic-reperfusion （IR） group （n=30）， Pertussis toxin （PT） group （n=30）. The blood flow of right common carotid artery of rat was blocked for 90 min to make ischemia reperfusion model. The levels of Gαi2 in hippocampus was assayed by immunohistochemistry and Western blotting after ischemia reperfu-sion for 6，12，24 h in each group. The average fluorescence values of intracellular Ca2+ levels in hippocampus of rats in three groups were detected by using Flow CytoMeter （FCM）. Neuronal cell apoptosis was measured by TUNEL.Results After restoration of middle cerebral artery blood flow for different lengths of time， the levels of hippocampus Gαi2 and Ca2+ levels in IR group were significantly higher than those in Sham group （P2+ levels in PT group were higher than those in IR group（P【Key words】Gαi2；Cerebral ischemia reperfusion injury；Hippocampus；Pertussis toxin；Apoptosis；Neuron；Ca2+；Death 脑缺血-再灌注损伤机制中，细胞内钙超载被认为是细胞死亡的最后共同通路，它促发多种病理损害过程，如炎症、自由基的形成、膜降解、线粒体功能紊乱等，最终介导细胞不可