title: Glycated high-density lipoprotein induces apoptosis of endothelial cells via a mitochondrial dysfunction.
abstract: Glycation of plasma proteins may contribute to an excess risk of developing atherosclerosis in patients with diabetes mellitus. Although it is believed that high-density lipoprotein (HDL) is nonenzymatically glycosylated at an increased level in diabetic individuals, little is known about a possible linkage between glycated HDL and endothelium dysfunction in diabetes. This study set out to clarify whether glucose-modified HDL affects the function of endothelial cells by examining the apoptosis of cultured human aortic endothelial cells (HAECs) exposed to a glycated-oxidized HDL (gly-ox-HDL) prepared in vitro. Incubation of HAECs with 100 microg/ml of gly-ox-HDL for 48 h showed apoptotic features, such as cell shrinkage, membrane blebbing, and concentration and fragmentation of the nucleus, and the degree of apoptosis was dose-dependent on the glucose used in the preparation of gly-ox-HDL. Stimulation of HAECs with gly-ox-HDL elicited a marked increase in caspase 3 activity and the expressions of active caspase 3 and caspase 9, whereas concomitant treatment with a caspase 3 inhibitor significantly blocked gly-ox-HDL-induced apoptosis of HAECs. The release of cytochrome c into cytosols markedly increased in HAECs during the treatment with gly-ox-HDL. The increased expressions of Bax and Bad were detected in HAECs incubated for 24 h with gly-ox-HDL, but gly-ox-HDL failed to interfere with the expression of Bcl-2 and Bcl-x. Moreover, in vitro experiments with HDL (gly-HDL) glycated in the presence of 2 mM EDTA and Cu(2+)-oxidized HDL suggested that the apoptotic effect of gly-ox-HDL on endothelial cells might be due to an additional oxidative modification of gly-HDL. Taken altogether, additional oxidation of HDL under hyperglycemic conditions may induce endothelial apoptosis through a mitochondrial dysfunction, following the deterioration of vascular function.
authors.affiliation: First Department of Biochemistry, Saitama Medical School, 38 Morohongo, Moroyama, Iruma-gun, Saitama, 350-0495, Japan. t_matsu@saitama-med.ac.jp
authors.forename: T
authors.initials: T
authors.lastname: Matsunaga
authors.forename: K
authors.initials: K
authors.lastname: Iguchi
authors.forename: T
authors.initials: T
authors.lastname: Nakajima
authors.forename: I
authors.initials: I
authors.lastname: Koyama
authors.forename: T
authors.initials: T
authors.lastname: Miyazaki
authors.forename: I
authors.initials: I
authors.lastname: Inoue
authors.forename: S
authors.initials: S
authors.lastname: Kawai
authors.forename: S
authors.initials: S
authors.lastname: Katayama
authors.forename: K
authors.initials: K
authors.lastname: Hirano
authors.forename: S
authors.initials: S
authors.lastname: Hokari
authors.forename: T
authors.initials: T
authors.lastname: Komoda
mesh_terms: D001011:Aorta / Q000166:cytology; D017209:Apoptosis; D015153:Blotting, Western; D002352:Carrier Proteins / Q000096:biosynthesis; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D003574:Cytochrome c Group / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism / Q000473:pathology*; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D008075:Lipoproteins, HDL / Q000378:metabolism* / Q000494:pharmacology; D008858:Microscopy, Phase-Contrast; D008928:Mitochondria / Q000378:metabolism*; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis; D019253:Proto-Oncogene Proteins c-bcl-2; D013997:Time Factors; D051028:bcl-2-Associated X Protein; D051025:bcl-Associated Death Protein
chemical_list: C494819:BAD protein, human; C494826:BAX protein, human; D002352:Carrier Proteins; D003574:Cytochrome c Group; D008075:Lipoproteins, HDL; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D051025:bcl-Associated Death Protein; C505407:CASP3 protein, human; C506433:CASP9 protein, human; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases; D005947:Glucose