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 kina...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。展开更多
Several factors have been implicated in obesity-related hypertension, but the genesis of the hypertension is largely unknown. In this study, we found a significantly upregulated expression of CPI-17(C-kinasepotentiate...Several factors have been implicated in obesity-related hypertension, but the genesis of the hypertension is largely unknown. In this study, we found a significantly upregulated expression of CPI-17(C-kinasepotentiated protein phosphatase 1 inhibitor of 17 kDa) and protein kinase C(PKC) isoforms in the vascular smooth muscles of high-fat diet(HFD)-fed obese mice. The obese wild-type mice showed a significant elevation of blood pressure and enhanced calcium-sensitized contraction of vascular smooth muscles. However, the obese CPI-17-deficient mice showed a normotensive blood pressure, and the calcium-sensitized contraction was consistently reduced. In addition, the mutant muscle displayed an abolished responsive force to a PKC activator and a 30%-50% reduction in both the initial peak force and sustained force in response to various G protein-coupled receptor(GPCR) agonists. Our observations showed that CPI-17-mediated calcium sensitization is mediated through a GPCR/PKC/CPI-17/MLCP/RLC signaling pathway. We therefore propose that the upregulation of CPI-17-mediated calcium-sensitized vasocontraction by obesity contributes to the development of obesity-related hypertension.展开更多
文摘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。
基金supported by the National Natural Science Funding of China (31272311 and 31330034 to M.S.Z.)
文摘Several factors have been implicated in obesity-related hypertension, but the genesis of the hypertension is largely unknown. In this study, we found a significantly upregulated expression of CPI-17(C-kinasepotentiated protein phosphatase 1 inhibitor of 17 kDa) and protein kinase C(PKC) isoforms in the vascular smooth muscles of high-fat diet(HFD)-fed obese mice. The obese wild-type mice showed a significant elevation of blood pressure and enhanced calcium-sensitized contraction of vascular smooth muscles. However, the obese CPI-17-deficient mice showed a normotensive blood pressure, and the calcium-sensitized contraction was consistently reduced. In addition, the mutant muscle displayed an abolished responsive force to a PKC activator and a 30%-50% reduction in both the initial peak force and sustained force in response to various G protein-coupled receptor(GPCR) agonists. Our observations showed that CPI-17-mediated calcium sensitization is mediated through a GPCR/PKC/CPI-17/MLCP/RLC signaling pathway. We therefore propose that the upregulation of CPI-17-mediated calcium-sensitized vasocontraction by obesity contributes to the development of obesity-related hypertension.
