CPI-17、ILK、ZIPK与PKCα、ε在低氧大鼠血管平滑肌中的相互作用  

The interaction between CPI-17,ILK,ZIPK and PKCα,PKCε in vascular smooth muscle of rats with hypoxia

作　　者：徐竞[1] 明佳[1] 李涛[1] 杨光明[1] 陈玮[1] 刘良明[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所二室创伤烧伤与复合伤国家重点实验室,重庆400042

出　　处：《创伤外科杂志》2009年第1期66-69,共4页Journal of Traumatic Surgery

基　　金：国家重大基础研究计划项目(2005CB522601);国家杰出青年科学基金(30625037)

摘　　要：目的本实验室前期研究发现蛋白激酶Cα(protein kinase Cα,PKCα)、蛋白激酶Cε(protein ki-nase Cε,PKCε)可能通过改变CPI-17(PKC-potentiated inhibitory protein for protein phosphatase 1 of 17kDa)、ILK(integrin-linked kinase)、ZIPK(zipper-interacting protein kinase)的蛋白表达和活性,来调节失血性休克后血管钙敏感性,但这些分子是否存在直接相互作用,尚不清楚。本实验观察CPI-17、ILK、ZIPK与PKCα、ε在低氧血管平滑肌细胞(vascualr smooth muscle cell,VSMC)中的相互作用。方法采用低氧培养的大鼠VSMC,运用免疫共沉淀技术,观察PKCα、ε与下游分子CPI-17、ILK、ZIPK,以及CPI-17、ILK、ZIPK之间的相互作用。结果(1)在PKCα抗体捕获的免疫沉淀中,ILK、ZIPK抗体,尤其是ILK抗体可杂交出明显蛋白条带,CPI-17抗体不能杂交出蛋白条带;(2)在PKCε抗体捕获的免疫沉淀中,ILK、ZIPK抗体均可杂交出明显蛋白条带,CPI-17抗体不能杂交出蛋白条带;(3)在CPI-17抗体捕获的免疫沉淀中,ILK、ZIPK抗体可杂交出明显蛋白条带;在ILK抗体捕获的免疫沉淀中,ZIPK抗体可杂交出明显蛋白条带。结论PKCα可直接作用于ILK,PKCε可直接作用于ILK和ZIPK,ILK、ZIPK可直接作用于CPI-17,ILK可直接作用于ZIPK。上述分子相互作用,形成网络,共同调节大鼠失血性休克后的血管钙敏感性。Objective Our previous research showed that protein kinase Co^（PKCα）, protein kinase Cε （PKCε） regulated vascular calcium sensitivity through PKC-potentiated inhibitory protein for protein phosphatase 1 of 17kDa（ CPI-17）,integrin-linked kinase（ILK） and zipper-interacting protein kinase（ZIPK）, while their interaction is not clear, so this research is to observe the interaction between CPI-17 ,ILK,ZIPK and PKCα,PKC, in vascu- lar smooth muscle eell（VSMC） after hypoxia. Methods The hypoxic VSMC of rats were adopted to observe the interaction between PKCα ,PKCε and the downstream molecular CPI-17, ILK,ZIPK,and the interaction among CPI-17, ILK, ZIPK through eo-immunopreeipitation. Results （ 1 ] ILK and ZIPK, especially ILK, were present in immunopreeipitates of PKCα,while CPI-17 was not observed. （2） ILK and ZIPK were both obviously present in immunoprecipitates of PKCε,while CPI-17 was not either. （3） ILK and ZIPK were both obviously present in immuno-precipitates of CPI-17, and ZIPK was present in immunopreeipitates of ILK. Conclusion PKCα interacts with ILK. PKC ε interacts with ILK and ZIPK. ILK and ZIPK interacts with CPI-17,ILK interacts with ZIPK. These molecules interact with each other and form a cooperative network contributing to regulation of vascular calcium sensitivity following hemorrhagic shock in rats.

关 键 词：失血性休克 钙敏感性 蛋白激酶CΑ 蛋白激酶CΕ CPI-17 ILK ZIPK 

分 类 号：R363[医药卫生—病理学]