title: Dual effect of oxidized LDL on cell cycle in human endothelial cells through oxidative stress.
abstract: BACKGROUND
  Oxidized low-density lipoprotein (OxLDL) exerts proliferation and apoptosis in vascular cells, depending on its concentration and the exposure time. Various steps in the cell cycle and in the apoptotic signaling cascade are modulated by O2-, and OxLDL stimulates vascular O2- formation. Here we studied the role of NADPH oxidase, a potential source for O2- formation after OxLDL stimulation, in cell proliferation, and we investigated whether OxLDL influences anti-apoptotic genes in cultured human umbilical vein endothelial cells (HUVEC). Methods and Results. OxLDL dose-dependently (10 to 300 microg/mL) stimulated O2- formation in HUVEC (detected by cytochrome c assay and by chemiluminescence of lucigenin). Low OxLDL concentrations (5 to 10 microg/mL) induced proliferation (detected by 3H-thymidine incorporation), while higher concentrations (50 to 300 microg/mL) induced apoptotic cell death (detected by Annexin assay and DNA fragmentation). Proliferation was blocked by the antioxidants SOD and catalase and by diphenyleneiodonium (10 micromol/L), an inhibitor of the O2- generating NADPH oxidase. In addition, cells transfected with antisense oligonucleotides for NADPH oxidase showed a significantly reduced O2- formation after stimulation with OxLDL. The OxLDL effect on apoptosis was also blocked by antioxidants. Since endothelial cells are protected against apoptosis through anti-apoptotic genes, we investigated whether OxLDL overcomes protection against apoptosis through suppression of the anti-apoptotic gene A20, a zinc-finger protein. OxLDL suppressed the expression of A20 in a dose-dependent manner.
  
  
  CONCLUSIONS
  These data indicate that OxLDL has a dual effect on cell cycle in HUVEC, inducing proliferation at low and apoptosis at higher concentrations. Both effects are mediated by O2- formation, with NADPH oxidase being a major source for O2-. Thus, OxLDL contributes importantly to vascular cellular turnover through the induction of oxidative stress.
authors.affiliation: Division of Nephrology, Department of Medicine, University Hospital of Würzburg, Würzburg, Germany. j-c.galle@mail.uni-wuerzburg.de
authors.forename: J
authors.initials: J
authors.lastname: Galle
authors.forename: A
authors.initials: A
authors.lastname: Heinloth
authors.forename: C
authors.initials: C
authors.lastname: Wanner
authors.forename: K
authors.initials: K
authors.lastname: Heermeier
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001483:Base Sequence; D002374:Catalase / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D047908:Intracellular Signaling Peptides and Proteins; D008077:Lipoproteins, LDL / Q000494:pharmacology*; D026901:Membrane Transport Proteins; D009252:NADPH Dehydrogenase / Q000037:antagonists & inhibitors / Q000235:genetics; D019255:NADPH Oxidases; D009687:Nuclear Proteins; D016376:Oligonucleotides, Antisense / Q000235:genetics; D009861:Onium Compounds / Q000494:pharmacology; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010750:Phosphoproteins / Q000037:antagonists & inhibitors / Q000235:genetics; D011506:Proteins / Q000235:genetics; D013482:Superoxide Dismutase / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism; D014162:Transfection; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008077:Lipoproteins, LDL; D026901:Membrane Transport Proteins; D009687:Nuclear Proteins; D016376:Oligonucleotides, Antisense; D009861:Onium Compounds; D010750:Phosphoproteins; D011506:Proteins; C099252:oxidized low density lipoprotein; D013481:Superoxides; C007517:diphenyleneiodonium; D002374:Catalase; D013482:Superoxide Dismutase; D019255:NADPH Oxidases; C086986:CYBA protein, human; D009252:NADPH Dehydrogenase; C076529:TNFAIP3 protein, human; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3