title: Oxidative modifications of low-density lipoproteins (LDL) by the human endothelial cell line EA.hy 926.
abstract: Modifications of LDL by the EA.hy 926 cell line were compared to those generated by human umbilical vein endothelial cells (HUVEC). Thiobarbituric acid reactive substances (TBARS) index values (TBARS sample/TBARS cell-free control ratio) were 2.64 +/- 0.18 (m +/- SE, n = 11) and 3.12 +/- 0.24 (n = 11), for HUVEC and EA.hy 926, respectively. The percentage of the most electronegative modified LDL fraction (fraction C), assessed by using an ion-exchange chromatographic method based on fast protein liquid chromatography (FPLC), represented 14 +/- 3% (n = 34) and 22 +/- 13% (n =10) of total modified LDL in HUVEC and EA.hy 926, respectively. LDL modified by both cell lines showed increased agarose electrophoretic mobility and apo B100 fragmentation on SDS-PAGE. None of the results were significantly different between the two cell lines. Superoxide anion production was 0.12 +/- 0.04 (n = 11) and 0.07 +/- 0.01 nmol/min/mg cell protein (n = 11) in HUVEC and EA.hy 926, respectively. Cell-specific effects on LDL were abrogated in cysteine-free medium. Moreover, cell-modified LDL were similarly degraded by J774 macrophage-like cells. We conclude that EA.hy 926 cells are a good model for investigating endothelial cell-induced modifications of LDL. Advantages include ready availability and less individual variability than with HUVEC.
authors.affiliation: Laboratoire de Biochimie Aplliquée, Faculté de Pharmacie, Châtenay-Malabry, France.
authors.forename: M A
authors.initials: MA
authors.lastname: Pech-Amsellem
authors.forename: I
authors.initials: I
authors.lastname: Myara
authors.forename: I
authors.initials: I
authors.lastname: Pico
authors.forename: C
authors.initials: C
authors.lastname: Mazière
authors.forename: J C
authors.initials: JC
authors.lastname: Mazière
authors.forename: N
authors.initials: N
authors.lastname: Moatti
mesh_terms: D002460:Cell Line; D003545:Cysteine / Q000378:metabolism; D003553:Cystine / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism; D010084:Oxidation-Reduction; D013481:Superoxides / Q000378:metabolism
chemical_list: D008077:Lipoproteins, LDL; D013481:Superoxides; D003553:Cystine; D003545:Cysteine