Atherogenic levels of low-density lipoprotein increase hydrogen peroxide generation in cultured human endothelial cells: possible mechanism of heightened endocytosis.
Cultured human umbilical vein endothelial cells(EC) exposed to atherogenic low-density lipoprotein (LDL) levels have augmented reactive oxygen species generation. Confluent EC were incubated with 30-330 mg/dl LDL cholesterol and cellular hydrogen peroxide (H2O2) generation measured. EC incubated with 30 and 90 mg/dl LDL cholesterol showed similar low level H2O2 production. In contrast, EC exposed to 180 and 330 mg/dl LDL cholesterol have a marked, dose-related elevation in H2O2 generation. Subsequent studies have explored if direct EC exposure to H2O2 promotes cellular functional changes similar to those induced by high LDL levels (> 160 mg/dl cholesterol). Confluent EC were incubated with 0.1-10 mM H2O2 for 30 minutes and endocytosis measured and cytoskeletal structure examined. H2O2 exposure (0.5 and 1 mM) promoted heightened EC endocytosis, which similarly occurs with high LDL exposure. Likewise, cytoskeletal examination of EC perturbed with 1 mM H2O2 reveals structural remodeling with a marked increase in stress fibers, which similarly happens with high LDL levels. The above observations that high LDL levels cause increased EC H2O2 production, and direct H2O2 exposure promotes cellular functional changes similar to those induced by high LDL concentrations, suggest a modulatory role for reactive oxygen species. Thus LDL-induced reactive oxygen species generation may contribute mechanistically to endothelial perturbation, which has been hypothesized to be a major contributing factor in the pathogenesis of atherosclerosis.
Holland
J A
JA
Department of Medicine, State University of New York Health Science Center, Syracuse 13210, USA.
Ziegler
L M
LM
Meyer
J W
JW
D000963:Antimetabolites / Q000494:pharmacology; D001161:Arteriosclerosis / Q000378:metabolism*; D019328:Buthionine Sulfoximine; D002478:Cells, Cultured / Q000166:cytology / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D008077:Lipoproteins, LDL / Q000494:pharmacology*; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D014471:Umbilical Veins / Q000166:cytology
D000963:Antimetabolites; D008077:Lipoproteins, LDL; D008717:Methionine Sulfoximine; D019328:Buthionine Sulfoximine; D006861:Hydrogen Peroxide