Glycated collagen I induces premature senescence-like phenotypic changes in endothelial cells.
Diabetic vasculopathy is central to the development of diverse cardiovascular, renal, retinal, and neurological complications of diabetes. We previously demonstrated that growth of endothelial cells on glycated extracellular matrix proteins (collagen and matrigel) results in a significant decrease in cell proliferation. In the present study, we show that early-passage human umbilical vein endothelial cells (HUVECs) grown on glycated collagen (GC) express hallmarks of premature cell senescence, ie, increase in the proportion of cells expressing senescence-associated beta-galactosidase activity, apoptotic rate, and p53 and p14(AFR) expression, but in contrast to replicative senescence, display neither attrition of telomeres nor decrease in telomerase activity. An increased frequency of prematurely senescent cells was similarly observed in vivo in aortae of young Zucker diabetic rats, compared with lean controls. NO production by HUVECs grown on GC was decreased, despite a 3-fold increase in eNOS expression and was associated with the increased nitrotyrosine-modified proteins. Development of premature senescence of HUVECs on GC could be prevented and reversed by treatments with the peroxynitrite scavenger, ebselen, eNOS intermediate N(omega)-hydroxy-L-arginine (NOHA), or superoxide dismutase mimetic Mn-TBAP. Concomitant with the reversal of senescence, ebselen, and NOHA each restored NO production to levels observed with HUVECs grown on unmodified collagen. Our findings indicate that diabetes mellitus in vivo and GC exposure in vitro elicit premature senescence of the vascular endothelium, a process with distinct pathogenetic mechanisms. Premature senescence of the vascular endothelium is hypothesized to be an important contributor to diabetic vasculopathy and a consequence of reduced NO availability, peroxynitrite, and/or superoxide excess.
Chen
Jun
J
Department of Medicine, State University of New York, Stony Brook, NY, USA.
Brodsky
Sergey V
SV
Goligorsky
David M
DM
Hampel
Dierk J
DJ
Li
Hong
H
Gross
Steven S
SS
Goligorsky
Michael S
MS
D000818:Animals; D017209:Apoptosis; D001120:Arginine / Q000031:analogs & derivatives* / Q000494:pharmacology; D001393:Azoles / Q000494:pharmacology; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000187:drug effects; D024042:Collagen Type I / Q000378:metabolism / Q000494:pharmacology*; D003921:Diabetes Mellitus, Experimental / Q000473:pathology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016166:Free Radical Scavengers / Q000494:pharmacology; D006031:Glycosylation; D006801:Humans; D054833:Isoindoles; D007700:Kinetics; D008665:Metalloporphyrins / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III; D016566:Organoselenium Compounds / Q000494:pharmacology; D030421:Peroxynitrous Acid / Q000378:metabolism; D010641:Phenotype; D051381:Rats; D011924:Rats, Zucker; D001616:beta-Galactosidase / Q000032:analysis
D001393:Azoles; D024042:Collagen Type I; D016166:Free Radical Scavengers; D054833:Isoindoles; D008665:Metalloporphyrins; D016566:Organoselenium Compounds; C097284:manganese(III)-tetrakis(4-benzoic acid)porphyrin; D030421:Peroxynitrous Acid; D009569:Nitric Oxide; C042986:ebselen; C068309:N(omega)-hydroxyarginine; D001120:Arginine; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat; D001616:beta-Galactosidase