title: Lipopolysaccharide induces inducible nitric oxide synthase-dependent podocyte dysfunction via a hypoxia-inducible factor 1α and cell division control protein 42 and Ras-related C3 botulinum toxin substrate 1 pathway.
abstract: Urine protein loss in immune complex-mediated diseases such as lupus nephritis is associated with podocyte foot process effacement (podocytopathy) but is not always dependent on glomerular immune complex deposition. Several murine and human studies have associated lupus nephritis with inducible nitric oxide synthase (iNOS) expression in what appear to be podocytes. This study was conducted to determine mechanisms of immune-complex-independent and iNOS-dependent podocyte dysfunction. Conditionally immortalized podocytes were cultured with lipopolysaccharide (LPS) and nitric oxide (NO), superoxide (SO), or peroxynitrite donors in the presence or absence of inhibitors of iNOS, reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase or monocyte chemotactic protein 1 (MCP-1), or with sepiapterin to increase coupling of iNOS homodimers. Podocyte NO, SO, and MCP-1 production and nitrotyrosine modifications were determined. The podocytopathy phenotype was determined by measuring cell motility and membrane permeability to albumin. This study determined that NO produced by iNOS is sufficient and necessary to induce podocytopathy. NO probably induces this phenotype via hypoxia-inducible factor 1α and cell division control protein 42 and Ras-related C3 botulinum toxin substrate 1 pathways. With LPS stimulation, neither SO nor peroxynitrite produced by uncoupled iNOS or NADPH oxidase nor MCP-1 was sufficient to induce the full phenotype. This study supports the notion that iNOS may induce autocrine podocyte dysfunction. Thus, targeting iNOS or the pathways of its induction may have therapeutic benefit.
authors.affiliation: Division of Rheumatology and Immunology, Department of Medicine, Medical University of South Carolina, Charleston, SC 29425, USA.
authors.forename: Ahmad K
authors.initials: AK
authors.lastname: Mashmoushi
authors.affiliation: Division of Rheumatology and Immunology, Department of Medicine, Medical University of South Carolina, Charleston, SC 29425, USA; Rheumatology Section, Medical Service, Ralph H. Johnson VA Medical Center, Charleston, SC 29401, USA. Electronic address: oatesjc@musc.edu.
authors.forename: Jim C
authors.initials: JC
authors.lastname: Oates
mesh_terms: D000818:Animals; D019898:Autocrine Communication; D002460:Cell Line; D002463:Cell Membrane Permeability; D002465:Cell Movement; D018932:Chemokine CCL2 / Q000096:biosynthesis; D004790:Enzyme Induction / Q000187:drug effects; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008822:Mice, Transgenic; D009479:Neuropeptides / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis; D052247:Nitric Oxide Synthase Type II / Q000235:genetics* / Q000378:metabolism; D050199:Podocytes / Q000201:enzymology* / Q000276:immunology; D055503:Protein Multimerization; D015398:Signal Transduction; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism*; D020830:rac1 GTP-Binding Protein / Q000378:metabolism*
chemical_list: C491768:Ccl2 protein, mouse; C581598:Cdc42 protein, mouse; D018932:Chemokine CCL2; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008070:Lipopolysaccharides; D009479:Neuropeptides; C484602:Rac1 protein, mouse; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein
keywords: Cell division control protein 42; Free radicals; Hypoxia-inducible factor 1α; Inducible nitric oxide synthase; Lipopolysaccharide; Lupus nephritis; Nitric oxide; Peroxynitrite; Podocyte; Ras-related C3 botulinum toxin substrate 1; Superoxide; Toll-like receptor 4