cPKCγ-HSP60信号通路参与小鼠脑低氧预适应  

cPKCγ-HSP60 signal pathway participates in hypoxic preconditioning in mice

作　　者：蒋淑君[1] 张楠[2] 李俊发[2] 

机构地区：[1]滨州医学院生理学教研室,山东烟台264003 [2]首都医科大学神经生物学系,北京100069

出　　处：《中国病理生理杂志》2011年第6期1066-1070,共5页Chinese Journal of Pathophysiology

基　　金：山东省卫生发展科技计划资助项目(No.2009HW006)

摘　　要：目的:鉴定参与低氧预适应(HPC)的常规型蛋白激酶Cγ(cPKCγ)相互作用蛋白,探讨与cPKCγ相互作用的热休克蛋白60(HSP60)表达水平在小鼠脑低氧预适应和脑缺血中的变化。方法:健康雄性BALB/c小鼠,随机分为:常氧组(Norm)和HPC组,结合是否进行大脑中动脉阻塞(MCAO)分为常氧假手术组(Norm+sham)、常氧缺血组(Norm+I)、HPC假手术组(HPC+sham)和HPC缺血组(HPC+I)。应用整体低氧预适应和MCAO缺血模型,结合免疫沉淀、双向凝胶电泳、质谱等技术分离、鉴定与cPKCγ相互作用的蛋白;利用蛋白印迹技术分析HSP60蛋白表达量在脑HPC和缺血中的变化。结果:与常氧组比较,HPC小鼠脑皮层膜相关成分中与cPKCγ相互作用的HSP60蛋白表达量升高,经免疫共沉淀证明cPKCγ与HSP60存在相互作用。在MCAO缺血模型中,与Norm+sham组小鼠相比,Norm+I组及HPC+I组小鼠皮层缺血核心区、半影区HSP60表达水平明显升高(P<0.05)。HPC+I组小鼠缺血核心区HSP60表达水平低于Norm+I组(P<0.05)。结论:研究结果表明,cPKCγ-HSP60信号通路可能参与了低氧预适应的发生发展过程。AIM：To identify the proteins interacted with conventional protein kinase Cγ（cPKCγ） which are involved in hypoxic preconditioning（HPC）,and to investigate the role of cPKCγ-interacted heat-shock protein 60（HSP60） in the development of HPC and ischemia（I）.METHODS： Healthy male BALB/c mice were randomly divided into normoxia（Norm） and HPC groups.Based on whether middle cerebral artery occlusion（MCAO） was performed,the mice were divided into Norm＋sham group,Norm＋I group,HPC＋sham group and HPC＋I group（all n=6）.Immunoprecipitation,two-dimensional electrophoresis and mass spectrometry were applied to identify the proteins interacted with cPKCγ.The changes of HSP60 expression in the brain of the mice after HPC and MCAO were analyzed by Western blotting.RESULTS： The interaction of cPKCγ and HSP60 was confirmed by co-immunoprecipitation result.Compared with Norm groups,the expression level of cPKCγ-interacted HSP60 obviously increased in particulate fraction of cerebral cortex in HPC mice.In the MCAO ischemic animals,the level of HSP60 expression was significantly higher in ischemic core and penumbra in Norm＋I group and HPC＋I group than that in Norm＋sham group.HSP60 expression in ischemic core was lower in HPC＋I group than that in Norm＋I group.CONCLUSION： cPKCγ-HSP60 signal pathway might be involved in the development of cerebral hypoxic preconditioning in MCAO mice.

关 键 词：常规型蛋白激酶Cγ 蛋白质组 热休克蛋白60 低氧预适应 大脑中动脉阻塞 

分 类 号：R339.5[医药卫生—人体生理学]