Literature DB >> 18651000

Immunohistochemical analysis of target proteins of Rho-kinase in a mouse model of accelerated atherosclerosis.

Mark Rekhter1, Kaarthik Chandrasekhar, Donetta Gifford-Moore, Xiao-di Huang, Pamela Rutherford, Jeffrey Hanson, Raymond Kauffman.   

Abstract

BACKGROUND: The pleiotropic antiatherosclerotic effects of statins are believed to be associated with the inhibition of Rho-kinase. However, a systematic analysis of Rho-kinase activation in atherosclerotic lesions is missing.
OBJECTIVES: To analyze the distribution and phosphorylation of target proteins of Rho-kinase, such as myosin light chain (MLC) and ezrin-radixin-moesin (ERM) proteins, in the apolipoprotein E (ApoE) knockout model of accelerated atherosclerosis, as well as the effects of treatment with the Rho-kinase inhibitor Y-27632.
METHOD: Western diet-fed ApoE-deficient mice underwent carotid ligation and were sacrificed 14 days after surgery. One group of ligated mice was treated with the Rho-kinase inhibitor Y-27632. Nonligated C57Bl6/J mice on normal chow and ApoE-deficient mice on Western diet were used as controls. Lesion structure and size were analyzed using Masson-elastic stained cross-sections. The distribution and phosphorylation of Rho-kinase target proteins were studied immunohistochemically.
RESULTS: Two weeks after surgery, atherosclerotic plaque-like lesions developed in ligated carotids. Lesion development was inhibited by Y-27632. ERM was expressed ubiquitously, but in the intact arteries, it was phosphorylated exclusively in the endothelium and periadventitial adipocytes. In the atherosclerotic lesions, foamy macrophages also exhibited a strong phospho-ERM signal. Y-27632 inhibited ERM phosphorylation in the plaques. MLC and phospho-MLC were associated with smooth muscle cells and did not respond to the Y-27632 treatment.
CONCLUSIONS: A cell type-selective distribution and phosphorylation of target proteins of Rho-kinase were demonstrated in the carotid artery of the normal mouse model, as well as in the ApoE-knockout model of accelerated atherosclerosis. Various downstream targets of the same enzyme may be differentially involved in specific pathological processes in a cell type-specific manner.

Entities:  

Keywords:  Atherosclerosis; Endothelium; Macrophages; Phosphorylation; Rho-kinase

Year:  2007        PMID: 18651000      PMCID: PMC2359611     

Source DB:  PubMed          Journal:  Exp Clin Cardiol        ISSN: 1205-6626


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