title: GD3 recruits reactive oxygen species to induce cell proliferation and apoptosis in human aortic smooth muscle cells.
abstract: Sialic acid containing glycosphingolipids (gangliosides) are expressed on the surface of all mammalian cells and have been implicated in regulating various biological phenomena; however, the detailed signaling mechanisms involved in this process are not known. We report here a novel aspect of disialoganglioside, GD3-mediated regulation of cell proliferation and cell death via the recruitment of reactive oxygen species (ROS). A low concentration (2.5-10 microm) of GD3, incubated with human aortic smooth muscle cells for a short period of time (10-30 min), stimulates superoxide generation via the activation of both NADPH oxidase and NADH oxidase activity. This leads to downstream signaling leading to cell proliferation and apoptosis. However, [(3)H]GD3 incubated with the cells under such conditions was found in a trypsin-sensitive fraction that was separable from endogenous GD3. The exact mechanism causing ROS generation and downstream signaling remains to be elucidated. The uptake of GD3 was accompanied by a 2.5-fold stimulation in the activity of mitogen-activated protein (MAP) kinase and 5-fold stimulation in cell proliferation. Preincubation of cells with membrane-permeable antioxidants, pyrrolidine dithiocarbamate, and N-acetylcysteine abrogated the superoxide generation and cell proliferation. In contrast, at higher concentrations (50-200 microm) GD3 inhibited the generation of superoxides but markedly stimulated the generation of nitric oxide (NO) (10-fold compared with control). This in turn stimulated mitochondrial cytochrome c release and intrachromosomal DNA fragmentation, which lead to apoptosis. In sum, at a low concentration, GD3 recruits superoxides to activate p44 MAPK and stimulates cell proliferation. In contrast, at high concentrations GD3 recruits nitric oxide to scavenge superoxide radicals that triggered signaling events that led to apoptosis. These observations might have relevance in regard to the potential role of GD3 in aortic smooth muscle cell proliferation and apoptosis that may contribute to plaque rupture in atherosclerosis.
authors.affiliation: Department of Pediatrics, Lipid Research Atherosclerosis Unit, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21044, USA.
authors.forename: Anil Kumar
authors.initials: AK
authors.lastname: Bhunia
authors.forename: Gunter
authors.initials: G
authors.lastname: Schwarzmann
authors.forename: Subroto
authors.initials: S
authors.lastname: Chatterjee
mesh_terms: D001011:Aorta / Q000166:cytology*; D017209:Apoptosis; D001161:Arteriosclerosis; D002455:Cell Division; D002478:Cells, Cultured; D003574:Cytochrome c Group / Q000378:metabolism; D053938:DNA Fragmentation; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology*; D005732:Gangliosides / Q000378:metabolism* / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D020935:MAP Kinase Signaling System; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009097:Multienzyme Complexes / Q000378:metabolism; D009130:Muscle, Smooth / Q000166:cytology*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D019255:NADPH Oxidases / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D011233:Precipitin Tests; D017382:Reactive Oxygen Species; D015398:Signal Transduction; D013347:Subcellular Fractions / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology
chemical_list: D003574:Cytochrome c Group; D005732:Gangliosides; D009097:Multienzyme Complexes; D017382:Reactive Oxygen Species; D013481:Superoxides; D009569:Nitric Oxide; C026226:ganglioside, GD3; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D014357:Trypsin