短暂脑缺血诱导大鼠海马PKC-ε膜转位及HSP70表达_医学论文.docVIP

短暂脑缺血诱导大鼠海马PKC-ε膜转位及HSP70表达_医学论文 短暂脑缺血诱导大鼠海马PKC-ε膜转位及HSP70表达_医学论文 【摘要】 目的 探讨蛋白激酶C-ε(protein kinase C, PKC-ε)和热休克蛋白 70(heat shock protein, HSP70)在脑缺血耐受形成过程中的作用。方法 将80只大鼠分为短暂脑缺血组和假手术组（n=40）,每大组根据取材时间又分为缺血后1 h、6 h、1 d、2 d、4 d 5个亚组（n=8）。应用大脑中动脉阻闭（middle cerebral artery occlusion, MCAO）方法和免疫印迹（western blot）技术观察大鼠短暂脑缺血20 min后海马神经细胞不同时间点PKC-ε膜转位及HSP70表达的变化。结果 假手术组各时间点的PKC-ε与HSP70无明显变化。在短暂脑缺血组，缺血后1 h海马神经细胞胞质PKC-ε的含量开始下降，至1天达最低点，持续到4天；而胞膜PKC-ε的变化趋势相反，含量于6 h显著增高，1天达到高峰，持续到4天；短暂缺血1 h后HSP70的表达无明显增加，6 h增多明显，1天达最高，持续至4天均处于较高水平。结论 短暂脑缺血诱导海马神经细胞PKC-ε膜转位和HSP70表达增加,可能是短暂脑缺血诱导缺血耐受的机制。 【关键词】 脑缺血 蛋白激酶 Abstract： Objective To investigate the roles of PKC-ε and HSP70 in the formation of brain ischemic tolerance. Methods Middle cerebral artery occlusion (MCAO) model and Western blot analysis were used to observe the membrane translocation of PKC-ε and the changes in HSP70 expression in the hippocampal neurons at different time points (1 h, 6 h, 1 d, 2 d and 4 d after a 20 min transient cerebral ischemia). The results were compared with those of the sham-operation group. Results No significant changes in PKC-ε and HSP70 were found in the sham-operation group. In the transient cerebral ischemic group, cytoplasm PKC-ε began to decrease 1 h after treatment, reached its lowest level on 1 d and remained at such a level until 4 d while the membrane PKC-ε tended to vary the opposite way: i.e. the membrane PKC-ε increased notably 6 h after transient ischemia, peaked on 1 d and remained high until 4 d. The expression of HSP70 began to increase obviously 1 h after transient ischemia, increased markedly at 6 h, peaked on 1 d and remained high until 4 d. Conclusion Transient ischemia induces membrane translocation of PKC-ε and increases the expression of HSP70, which may be the genetic mechanism of ischemic tolerance induced by transient ischemia. Key words: cerebral ischemia protein kinase C-ε heat shock proteins 70；ischemic tolerance 缺血预处理可以提高组织和器官对致死性缺血损伤的抵御能力。短暂缺血是预处理的手段，它可以调动体内一系列内源性保护机制，从而实