title: A novel nontoxic inhibitor of the activation of NADPH oxidase reduces reactive oxygen species production in mouse lung.
abstract: 1-Hexadecyl-3-trifluoroethylglycero-sn-2-phosphomethanol (MJ33) is a fluorinated phospholipid analog that inhibits the phospholipase A2 (PLA2) activity of peroxiredoxin 6 (Prdx6). Prdx6 PLA2 activity is required for activation of NADPH oxidase 2 and subsequent generation of reactive oxygen species (ROS). In vitro, MJ33 inhibited agonist-stimulated production of ROS by the isolated perfused mouse lung, lung microvascular endothelial cells, and polymorphonuclear leukocytes. MJ33 (0.02-0.5 µmol MJ33/kg body weight) in mixed unilamellar liposomes was administered to C57BL/6 mice by either intratracheal (i.t.) or i.v. routes. Lung MJ33 content, measured by liquid chromatography/mass spectroscopy, showed uptake of 67-87% of the injected dose for i.t. and 23-42% for i.v. administration at 4 hours postinjection. PLA2 activity of lung homogenates was markedly inhibited (>85%) at 4 hours postadministration. Both MJ33 content and PLA2 activity gradually returned to near control levels over the subsequent 24-72 hours. Mice treated with MJ33 at 12.5-25 µmol/kg did not show changes (compared with control) in clinical symptomatology, body weight, hematocrit, and histology of lung, liver, and kidney during a 30- to 50-day observation period. Thus, the toxic dose of MJ33 was >25 µmol/kg, whereas the PLA2 inhibitory dose was approximately 0.02 µmol/kg, indicating a high margin of safety. MJ33 administered to mice prior to lung isolation markedly reduced ROS production and tissue lipid and protein oxidation during ischemia followed by reperfusion. Thus, MJ33 could be useful as a therapeutic agent to prevent ROS-mediated tissue injury associated with lung inflammation or in harvested lungs prior to transplantation.
authors.affiliation: Institute for Environmental Medicine, University of Pennsylvania, 3620 Hamilton Walk, 1 John Morgan Building, Philadelphia, PA 19104, USA.
authors.forename: Intae
authors.initials: I
authors.lastname: Lee
authors.forename: Chandra
authors.initials: C
authors.lastname: Dodia
authors.forename: Shampa
authors.initials: S
authors.lastname: Chatterjee
authors.forename: John
authors.initials: J
authors.lastname: Zagorski
authors.forename: Clementina
authors.initials: C
authors.lastname: Mesaros
authors.forename: Ian A
authors.initials: IA
authors.lastname: Blair
authors.forename: Sheldon I
authors.initials: SI
authors.lastname: Feinstein
authors.forename: Mahendra
authors.initials: M
authors.lastname: Jain
authors.forename: Aron B
authors.initials: AB
authors.lastname: Fisher
mesh_terms: D000818:Animals; D001835:Body Weight / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005994:Glycerophosphates / Q000494:pharmacology* / Q000633:toxicity; D006400:Hematocrit; D066298:In Vitro Techniques; D007275:Injections, Intravenous; D007278:Injections, Spinal; D008081:Liposomes / Q000737:chemistry; D008168:Lung / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors* / Q000235:genetics; D018384:Oxidative Stress / Q000187:drug effects; D054467:Phospholipases A2 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D005994:Glycerophosphates; D008081:Liposomes; D008562:Membrane Glycoproteins; D017382:Reactive Oxygen Species; C078587:1-hexadecyl-3-trifluoroethylglycero-sn-2-phosphomethanol; C488842:Cybb protein, mouse; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D054467:Phospholipases A2