Extracellular matrix remodelling of endotheual cellwas induced by very low denstiy lipoproteins through NFKb activation

Predisposing risk factors for atherosclerosis, which include hypertension, diabetes, smoking, and hypercholesterolemia, are all linked to endothelial cell (EC) dysfunction (1). However, the remodelling of the vascular extracellular matrix (vECM) and the signaling cascade involved in EC dysfunction induced by normal very low density lipoprotein (N-VLDL)injury is still unclear (2). The aim of the present work was to study the remodelling of vECM and signaling of Human Umbilical Vein Endothelial Cells (HUVEC) in response to human N-VLDL.All the protocols performed were approved by the Bioethics Committee of the University of Buenos Aires, Argentina. Human N-VLDLwere isolated by ultracentrifugation from healthy volunteers. EC were obtained and cultured as described Ulrich-Merzenic (3). After reaching 70-80%confluence, HUVEC were incubated with 75 mcg/mL of N-VLDLin serum-free medium for 24 hs. After treatment, vECM remodelling was studied through: 1) proteoglycans core protein production, specifically decorin, versican and perlecan analysis by Western blot;2) glycosaminoglycans content studied by reverse phase HPLC; 3) matrix metalloproteinase (MMP) activity analyzed by zymography.