OGD增加小鼠离体海马组织内cPKCβII和nPKCε的膜转位  

Oxygenglucose deprivation increase the levels of cPKCβII and nPKCε membrane translocation in hippocampus slices of mice

作　　者：李华[1] 贾军[2] 韩松[1] 李俊发[1] 

机构地区：[1]首都医科大学基础医学院神经生物学系 [2]首都医科大学基础医学院生理学系,北京100054

出　　处：《中国临床解剖学杂志》2005年第6期635-638,共4页Chinese Journal of Clinical Anatomy

基　　金：国家自然科学基金(30270508;30470650);北京市自然科学基金(7032005);教育部优秀青年教师资助项目;北京市教委科技发展计划项目(KM200310025100)

摘　　要：目的:通过观察无糖低氧(OGD)刺激对小鼠海马脑片内蛋白激酶C(PKC)特定亚型膜转位水平(激活程度)的影响,进一步证实我们在整体低氧预适应小鼠模型上所获实验结果,并为离体海马脑片缺血/低氧预适应(I/HPC)模型建立及后续药物干预实验打下基础。方法:急性分离小鼠海马组织、制备400!μm厚度的脑片,并用无糖低氧(OGD)人工脑脊液(ACSF)模拟缺血/低氧刺激;应用SDS-PAGE和Westernbolt等生化技术,并结合GelDoc凝胶成像系统,半定量检测OGD刺激2、5、10、15和30min后海马脑片内cPKCβII和nPKCε的膜转位水平。结果:OGD刺激可增高海马脑片内cPKCβII和nPKCε的膜转位水平,且这种增高从刺激5min开始持续至30min均有显著差异(P<0.001,n=6)。结论:实验结果进一步证实cPKCβII和nPKCε可能参与脑缺血/低氧性预适应的形成过程,并提示OGD10min作为制备离体海马脑片I/HPC模型的预处理刺激较为合理。Objective：In order to confirm our previous findings in the brain of hypoxic preconditioned mice and establish the ischemic/hypoxic preconditioning model of organotypic hippocampal slices in vitro for the later drug interference, we observed the effects of oxygen/glucose deprivation （OGD） on levels of cPKCβⅡ and nPKCε membrane translocation. Methods： Hippocampus isolated from decapitated mice was prepared slices with 400μm thickness. The biochemistry techniques of SDS-PAGE and Western bolt were applied to determine the levels ofcPKCβⅡ and nPKCe membrane translocation in hippocampus slices at 0, 2, 5, 10, 15 and 30 min of OGD stimulation. Results： It was showed that levels of cPKC[3II and nPKCe membrane translocation were increased in response to 0-30 min OGD, and the statistic significance （P〈0.001, n=6） could be seen from 5 to 30 min OGD for both cPKCβⅡ and nPKCε. Conclusions： These results further confirmed that the activation of cPKCβⅡ and nPKCε may play an important role in the onset of ischemic/hypoxic preconditioning. It also suggests that the stimulation of 10 min OGD might be a proper pretreatment to induce ischemic/hypoxic preconditioning in hippocampus slices.

关 键 词：脑低氧预适应 蛋白激酶C 膜转位 海马脑片 

分 类 号：R339.5[医药卫生—人体生理学]