title: In vivo and in vitro evidence for the glycoxidation of low density lipoprotein in human atherosclerotic plaques.
abstract: Although there have been suggestions that the glycation and oxidation of low density lipoprotein (LDL) might increase its atherogenic potential, little is known about the presence of glycoxidative LDL in human atherosclerotic lesions. We developed specific antibodies against different immunological epitopes of AGE structures, including N(epsilon)-(carboxymethyl)lysine-protein adduct (CML), a glycoxidation product, and structure(s) other than CML (nonCML), and a monoclonal antibody against oxidized phosphatidylcholine (oxPC), as an epitope of oxidized LDL. Immunohistochemical analysis demonstrated that the CML- and oxPC-epitopes were accumulated mainly in macrophage-derived foam cells in atherosclerotic lesions, including fatty streaks and atherosclerotic plaques. On the other hand, the nonCML-epitope and apolipoprotein B were localized mainly in extracellular matrices of atherosclerotic lesions. The CML- and oxPC-epitopes were characterized by a model antigen-generating system using the copper ion-induced peroxidation and/or glucose-induced glycation of LDL. The glycoxidation of LDL caused the formation of CML-epitope with increasing concentrations of copper ion and glucose. It was also formed to some extent in LDL incubated with high concentrations (500 mM) of glucose. However, no CML-epitope was observed in oxidized LDL induced by copper ion alone. On the other hand, the formation of oxPC-epitope in LDL was dependent on copper ion-induced peroxidation, but independent of glucose-induced glycation. The addition of chelators, ethylenediaminetetraacetic acid and diethylenetriaminepentaacetic acid, reduced the increase in electrophoretic mobility and TBARS caused by the peroxidation and glycoxidation of LDL, but had no effects on the formation of fructosamine caused by the glycation and glycoxidation of LDL. Chelators as well as aminoguanidine protected the formation of CML-epitope in glycated or glycoxidative LDL. Although the formation of oxPC-epitope was completely inhibited by the addition of chelators, it was partially protected by aminoguanidine. These in vitro results suggest that the glycoxidative modification of LDL may occur in the arterial intima, and may contribute to the development of human atherosclerotic lesions.
authors.affiliation: Second Department of Pathology, School of Medicine, Fukuoka University, 45-1, 7-chome Nanakuma, Jonan-ku, 814-0180, Fukuoka, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Imanaga
authors.forename: N
authors.initials: N
authors.lastname: Sakata
authors.forename: S
authors.initials: S
authors.lastname: Takebayashi
authors.forename: A
authors.initials: A
authors.lastname: Matsunaga
authors.forename: J
authors.initials: J
authors.lastname: Sasaki
authors.forename: K
authors.initials: K
authors.lastname: Arakawa
authors.forename: R
authors.initials: R
authors.lastname: Nagai
authors.forename: S
authors.initials: S
authors.lastname: Horiuchi
authors.forename: H
authors.initials: H
authors.lastname: Itabe
authors.forename: T
authors.initials: T
authors.lastname: Takano
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000911:Antibodies, Monoclonal; D001018:Aortic Diseases / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D001161:Arteriosclerosis / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D002614:Chelating Agents / Q000494:pharmacology; D002648:Child; D002675:Child, Preschool; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005260:Female; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D006031:Glycosylation / Q000187:drug effects; D006146:Guanidines / Q000494:pharmacology; D006801:Humans; D007223:Infant; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008239:Lysine / Q000031:analogs & derivatives* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D010713:Phosphatidylcholines / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D004791:Enzyme Inhibitors; D000939:Epitopes; D017127:Glycation End Products, Advanced; D006146:Guanidines; D008077:Lipoproteins, LDL; D010713:Phosphatidylcholines; D017392:Thiobarbituric Acid Reactive Substances; C048496:N(6)-carboxymethyllysine; D019001:Nitric Oxide Synthase; D008239:Lysine; C004479:pimagedine