title: Physicochemical changes in human high-density lipoproteins (HDL) oxidized by gamma radiolysis-generated oxyradicals. Effect on their cholesterol effluxing capacity.
abstract: This paper describes an oxidative process of human high-density lipoproteins (HDL) based upon the action of oxygenated free radicals produced by water radiolysis (OH. and OH./O2.- free radicals at pH 7), monitored by both biochemical and physical markers. Classical biochemical markers (vitamin E, thiobarbituric acid-reactive substances (TBARS), conjugated dienes and differential fluorescence) were studied as a function of the radiation dose (from 0 to 800 Gy; dose rate = 2.7 x 10(-2) Gy.s(-1)). The fluorescence polarization anisotropy (r) was measured with 1,6-diphenylhexatriene (DPH). Vitamin E decrease and formation of lipid peroxidation products (thiobarbituric acid-reactive substances and conjugated dienes) were concomitant in the case of OH. free radicals alone, whereas these products appeared after a small threshold dose when OH. and O2.- free radicals were simultaneously produced in solution. At high radiation doses, TBARS concentrations have reached plateau values (approx. 2 or 7 nmol/mg lipid with OH. or OH./O2.- free radicals, respectively) which were much lower than those obtained after copper oxidation (approx. 15 or 29 nmol/mg lipid after 12 and 24 h incubation, respectively). The free radical-induced oxidative process has led to a rigidification of the HDL and was associated with low values of cholesterol effluxing capacities when these oxidized HDL were incubated with cholesterol-loaded human fibroblasts. Similar results were obtained with copper-oxidized HDL, under our experimental conditions. Consequently, these two kinds of oxidative modification of HDL resulted both in a loss of their capacity to remove cellular cholesterol, which could be explained by the fact that this ability was under the dependence of a HDL optimum fluidity.
authors.affiliation: Laboratoire de Biochimie, Hôpital de la Salpêtrière, Paris, France.
authors.forename: D
authors.initials: D
authors.lastname: Bonnefont-Rousselot
authors.forename: C
authors.initials: C
authors.lastname: Motta
authors.forename: A O
authors.initials: AO
authors.lastname: Khalil
authors.forename: R
authors.initials: R
authors.lastname: Sola
authors.forename: A E
authors.initials: AE
authors.lastname: La Ville
authors.forename: J
authors.initials: J
authors.lastname: Delattre
authors.forename: M
authors.initials: M
authors.lastname: Gardès-Albert
mesh_terms: D002784:Cholesterol / Q000378:metabolism*; D005454:Fluorescence Polarization; D005609:Free Radicals; D006801:Humans; D015227:Lipid Peroxidation; D008075:Lipoproteins, HDL / Q000737:chemistry / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000737:chemistry*; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis; D014810:Vitamin E / Q000032:analysis
chemical_list: D005609:Free Radicals; D008075:Lipoproteins, HDL; D017382:Reactive Oxygen Species; D017392:Thiobarbituric Acid Reactive Substances; D014810:Vitamin E; D002784:Cholesterol