The myristoylated alanine-rich C kinase substrate differentially regulates kinase interacting with stathmin in vascular smooth muscle and endothelial cells and potentiates intimal hyperplasia formation

摘要: Abstract Objective Restenosis limits the durability of all cardiovascular reconstructions. Vascular smooth muscle cell (VSMC) proliferation drives this process, but an intact, functional endothelium is necessary for vessel patency. Current strategies to prevent restenosis employ antiproliferative agents that affect both VSMCs and endothelial cells (ECs). Knockdown myristoylated alanine-rich C kinase substrate (MARCKS) arrests VSMC paradoxically potentiates EC proliferation. MARCKS knockdown decreases expression interacting with stathmin (KIS), increasing p27kip1 expression, arresting Here, we seek determine how influences KIS protein in these two types. Methods Primary human coronary artery ECs were used in vitro experiments. was depleted by transfection small interfering RNA. Messenger RNA quantitated real-time reverse transcription polymerase chain reaction. Protein determined Western blot analysis. Ubiquitination immunoprecipitation. binding assessed co-immunoprecipitation. Intimal hyperplasia induced CL57/B6 mice a femoral wire injury. knocked down in vivo application 10 μM targeting suspended 30% Pluronic F-127 gel. formation measurement intimal thickness on cross sections injured artery. Re-endothelialization quantitating Evans blue dye Results did not messenger either type. In presence cycloheximide, decreased stability had no effect ECs. The abrogated 26s proteasome inhibitor MG-132. binds significantly increased level ubiquitinated resulted expression. Furthermore, 5-ethynyl-2′-deoxyuridine integration reduced thickening. enhanced barrier function recovery 4 days after Conclusions differentially regulates difference due differential ubiquitination interaction provides possible explanation observed ubiquitination. persists in vivo, endothelium, abrogates formation.