title: Disruption of asymmetric dimethylarginine-induced RelA/P65 association with actin in endothelial cells.
abstract: Asymmetric dimethylarginine (ADMA) activates nuclear factor (NF)-κB in endothelial cells, while actin-stabilizing or -destabilizing drugs prevent ADMA-induced activation of NF-κB. Here we investigated how actin-targeting drugs regulated ADMA-induced NF-κB activation in endothelial cells. Human umbilical vein endothelial cells were treated with ADMA for 24 h in the absence and presence of cytochalasin D or jasplakinolide. Expression levels of proteins and genes were measured by immunoblotting and reverse-transcription polymerase chain reaction, respectively. Chromatin immunoprecipitation was used to detect the binding of NF-κB to the vascular cell adhesion molecule 1 (VCAM-1) promoter. The association of actin with RelA/P65 was detected by immunoprecipitation. It was demonstrated that ADMA induced IκBα degradation, increased nuclear RelA/P65 translocation, and promoted the binding of NF-κB to the VCAM-1 promoter. Consequently, this increased the expression of VCAM-1. In parallel studies, actin-stabilizing and -destabilizing drugs decreased ADMA-induced RelA/P65 nuclear translocation, interfered with NF-κB binding to the VCAM-1 promoter and prevented the expression of VCAM-1. This was independent of total RelA/P65 levels and ADMA-induced IκBα degradation. Most importantly, the association of RelA/P65 with actin was increased after stimulation with ADMA, and impaired after treatment with actin-targeting drugs. In brief, actin-stabilizing or -destabilizing drugs interfere with the ADMA-induced association of RelA/P65 with actin, and consequently disrupt NF-κB activation.
authors.affiliation: Department of Nephrology,  Affiliated Beijing Friendship Hospital,  Faculty of Kidney Diseases,  Capital Medical University, Beijing 100050,  China.
authors.forename: Weikang
authors.initials: W
authors.lastname: Guo
authors.forename: Dongliang
authors.initials: D
authors.lastname: Zhang
authors.forename: Liyan
authors.initials: L
authors.lastname: Wang
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Wenhu
authors.initials: W
authors.lastname: Liu
mesh_terms: D000199:Actins / Q000378:metabolism*; D021581:Active Transport, Cell Nucleus; D001120:Arginine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology*; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007249:Inflammation; D016328:NF-kappa B / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D013997:Time Factors; D051996:Transcription Factor RelA / Q000378:metabolism*; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
chemical_list: D000199:Actins; D016328:NF-kappa B; C498143:RELA protein, human; D051996:Transcription Factor RelA; D019010:Vascular Cell Adhesion Molecule-1; C018524:N,N-dimethylarginine; D001120:Arginine