title: Role of Myeloperoxidase Oxidants in the Modulation of Cellular Lysosomal Enzyme Function: A Contributing Factor to Macrophage Dysfunction in Atherosclerosis?
abstract: Low-density lipoprotein (LDL) is the major source of lipid within atherosclerotic lesions. Myeloperoxidase (MPO) is present in lesions and forms the reactive oxidants hypochlorous acid (HOCl) and hypothiocyanous acid (HOSCN). These oxidants modify LDL and have been strongly linked with the development of atherosclerosis. In this study, we examined the effect of HOCl, HOSCN and LDL pre-treated with these oxidants on the function of lysosomal enzymes responsible for protein catabolism and lipid hydrolysis in murine macrophage-like J774A.1 cells. In each case, the cells were exposed to HOCl or HOSCN or LDL pre-treated with these oxidants. Lysosomal cathepsin (B, L and D) and acid lipase activities were quantified, with cathepsin and LAMP-1 protein levels determined by Western blotting. Exposure of J774A.1 cells to HOCl or HOSCN resulted in a significant decrease in the activity of the Cys-dependent cathepsins B and L, but not the Asp-dependent cathepsin D. Cathepsins B and L were also inhibited in macrophages exposed to HOSCN-modified, and to a lesser extent, HOCl-modified LDL. No change was seen in cathepsin D activity or the expression of the cathepsin proteins or lysosomal marker protein LAMP-1. The activity of lysosomal acid lipase was also decreased on treatment of macrophages with each modified LDL. Taken together, these results suggest that HOCl, HOSCN and LDL modified by these oxidants could contribute to lysosomal dysfunction and thus perturb the cellular processing of LDL, which could be important during the development of atherosclerosis.
authors.affiliation: The Heart Research Institute, Sydney, New South Wales, Australia.|Sydney Medical School, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Fahd O
authors.initials: FO
authors.lastname: Ismael
authors.affiliation: The Heart Research Institute, Sydney, New South Wales, Australia.|Sydney Medical School, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Tessa J
authors.initials: TJ
authors.lastname: Barrett
authors.affiliation: The Heart Research Institute, Sydney, New South Wales, Australia.
authors.forename: Diba
authors.initials: D
authors.lastname: Sheipouri
authors.affiliation: The Heart Research Institute, Sydney, New South Wales, Australia.|Sydney Medical School, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Bronwyn E
authors.initials: BE
authors.lastname: Brown
authors.affiliation: The Heart Research Institute, Sydney, New South Wales, Australia.|Sydney Medical School, University of Sydney, Sydney, New South Wales, Australia.|Department of Biomedical Sciences, Panum Institute, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Davies
authors.affiliation: The Heart Research Institute, Sydney, New South Wales, Australia.|Sydney Medical School, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Clare L
authors.initials: CL
authors.lastname: Hawkins
mesh_terms: D000818:Animals; D050197:Atherosclerosis / Q000201:enzymology* / Q000473:pathology; D002403:Cathepsins / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D006997:Hypochlorous Acid / Q000378:metabolism; D008049:Lipase / Q000378:metabolism*; D008077:Lipoproteins, LDL / Q000378:metabolism; D051907:Lysosomal Membrane Proteins / Q000378:metabolism; D008247:Lysosomes / Q000201:enzymology* / Q000473:pathology; D008264:Macrophages / Q000201:enzymology* / Q000473:pathology; D051379:Mice; D009195:Peroxidase / Q000378:metabolism*; D013861:Thiocyanates / Q000378:metabolism; D052118:Lysosomal-Associated Membrane Protein 1
chemical_list: D002403:Cathepsins; D006997:Hypochlorous Acid; D008049:Lipase; D008077:Lipoproteins, LDL; D051907:Lysosomal Membrane Proteins; D009195:Peroxidase; D013861:Thiocyanates; C052255:Lamp1 protein, mouse; D052118:Lysosomal-Associated Membrane Protein 1; C014607:hypothiocyanite ion