title: Increased uptake of LDL by oxidized macrophages is the result of an initial enhanced LDL receptor activity and of a further progressive oxidation of LDL.
abstract: Iron ions were recently shown to induce cellular lipid peroxidation in macrophages, and these oxidized cells can convert native low-density lipoprotein (LDL) to oxidized LDL (Ox-LDL). The present study demonstrates that deoxycholic acid (DCA) and angiotensin II (ANG-II) can also induce oxidative modification of macrophages via metal ions independent mechanisms. Furthermore, incubation of LDL (200 micrograms of protein/ml) for 24 h at 37 degrees C with DCA, ANG-II, as well as FeSO4-induced oxidized macrophages, resulted in oxidative modification of the lipoprotein as evidenced by increased TBARS formation in LDL (by 50, 105, and 258%, respectively), decreased TNBS reactivity (by 45, 56, and 42%, respectively), and increased cellular uptake (by 60, 166, and 230%, respectively). A positive correlation (n = .88) was found between the extent of the cellular lipid peroxidation and the increment in the cellular uptake of the LDL. The oxidative modification of LDL by oxidized macrophages was found to be a progressive process. Incubation of LDL with oxidized macrophages for increasing periods of time up to 24 h resulted in progressive increment in: (1) the electrophoretic mobility of the LDL; (2) the TBARS formation in LDL; (3) the cellular uptake of LDL by the oxidized macrophages via the Ox-LDL receptor. Upon fractionation on a heparin-sepharose column of LDL that was incubated for different periods of time with oxidized macrophages, a gradual increment in the unbound LDL fraction was obtained, up to 72% after 24 h of incubation. During the first hour of LDL incubation with the oxidized macrophages a twofold increase in the cellular uptake of LDL by these cells was detected, although no significant oxidation of the lipoprotein occurred during this short time period. This effect could be attributed to an increased number of LDL receptors on the cell surface of the oxidized macrophages. In conclusion, increased uptake of LDL by oxidized macrophages results from two routes: (1) enhanced uptake via the LDL receptor due to increased LDL receptor activity; (2) lipoprotein uptake via the Ox-LDL receptors due to cellular modification of LDL. Both of these processes lead to macrophage cholesterol accumulation and foam cell formation, and thus contribute to accelerated atherosclerosis under oxidative stress.
authors.affiliation: Lipid Research Laboratory, Rambam Medical Center, Bruce Rappaport Faculty of Medicine, Technion, Israel.
authors.forename: B
authors.initials: B
authors.lastname: Fuhrman
authors.forename: O
authors.initials: O
authors.lastname: Judith
authors.forename: S
authors.initials: S
authors.lastname: Keidar
authors.forename: L
authors.initials: L
authors.lastname: Ben-Yaish
authors.forename: M
authors.initials: M
authors.lastname: Kaplan
authors.forename: M
authors.initials: M
authors.lastname: Aviram
mesh_terms: D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D017077:Culture Media, Conditioned; D003840:Deoxycholic Acid / Q000494:pharmacology; D004588:Electrophoresis, Cellulose Acetate; D005296:Ferrous Compounds / Q000494:pharmacology; D006493:Heparin / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D011973:Receptors, LDL / Q000276:immunology / Q000378:metabolism*; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis; D014810:Vitamin E / Q000494:pharmacology
chemical_list: D017077:Culture Media, Conditioned; D005296:Ferrous Compounds; D008077:Lipoproteins, LDL; D016877:Oxidants; D011973:Receptors, LDL; D017392:Thiobarbituric Acid Reactive Substances; C099252:oxidized low density lipoprotein; D003840:Deoxycholic Acid; D000804:Angiotensin II; D014810:Vitamin E; C020748:ferrous sulfate; D006493:Heparin