我们前期研究已证实在IPC保护作用中，14-3-3η可与PKCε相互作用，帮助后者向线粒体迁移，发挥心肌保护作用。本项目综合前人与自身研究结果，探讨在上述过程后，14-3-3η/PKCε复合物解离的方式。拟首先在cos7细胞中用免疫共沉淀、GST-pulldown和双标记免疫荧光法初步确认Efp、14-3-3η和泛素之间的相互作用；继而观察在心肌细胞IPC过程中高表达（≥正常表达200%）、低表达（≥正常表达50%）和正常水平Efp对泛素化及14-3-3η/PKCε复合物解离的影响；探讨IPC诱导的Efp上调对下游靶蛋白的调控是否领带于雌激素；并在整体动物水平验证以上实验结果，观测Efp对IPC保护效应的影响。为完善人们对IPC保护作用中14-3-3η信号路径的认识，揭示Efp在心肌细胞中的作用，以期发现新的药物作用靶点，为进一步开发新型心肌保护药物奠定良好的理论与实验基础。

Our previous studies have shown that PKCε is targeted mitochondria by 14-3-3η which play a protective role in IPC. This project is designed to the mechanism of 14-3-3η/PKCε complex dissociation after IPC. We will assess the interactions between Efp, 14-3-3η and ubiquitin using co-immunoprecipitation, GST-pulldown and double labeling immunofluorescence. Then we'll compare differences in ubiquitinate dissociation of 14-3-3η/PKCε complex at high (≥200% of normal level), low (≤ 50% of normal level ) and levels of Efp in IPC. In addition, we will investigate whether IPC-induced Efp increase is accompanied by changes in downstream targets and whether this is depend upon estrogen. Finally, we'll test if the in vitro results can be reproduced in animals. This project will be instrumental to our understanding of 14-3-3η pathway in IPC and identify the role of Efp in cardiac myocytes. It will also identify new drug target for myocardial protection.