title: Oxalate-induced activation of PKC-alpha and -delta regulates NADPH oxidase-mediated oxidative injury in renal tubular epithelial cells.
abstract: Oxalate-induced oxidative stress contributes to cell injury and promotes renal deposition of calcium oxalate crystals. However, we do not know how oxalate stimulates reactive oxygen species (ROS) in renal tubular epithelial cells. We investigated the signaling mechanism of oxalate-induced ROS formation in these cells and found that oxalate significantly increased membrane-associated protein kinase C (PKC) activity while at the same time lowering cytosolic PKC activity. Oxalate markedly translocated PKC-alpha and -delta from the cytosol to the cell membrane. Pretreatment of LLC-PK1 cells with specific inhibitors of PKC-alpha or -delta significantly blocked oxalate-induced generation of superoxide and hydrogen peroxide along with NADPH oxidase activity, LDH release, lipid hydroperoxide formation, and apoptosis. The PKC activator PMA mimicked oxalate's effect on oxidative stress in LLC-PK1 cells as well as cytosol-to-membrane translocation of PKC-alpha and -delta. Silencing of PKC-alpha expression by PKC-alpha-specific small interfering RNA significantly attenuated oxalate-induced cell injury by decreasing hydrogen peroxide generation and LDH release. We believe this is the first demonstration that PKC-alpha- and -delta-dependent activation of NADPH oxidase is one of the mechanisms responsible for oxalate-induced oxidative injury in renal tubular epithelial cells. The study suggests that the therapeutic approach might be considered toward attenuating oxalate-induced PKC signaling-mediated oxidative injury in recurrent stone formers.
authors.affiliation: Vattikuti Urology Institute, Henry Ford Health System, Detroit, Michigan 48202, USA.
authors.forename: Vijayalakshmi
authors.initials: V
authors.lastname: Thamilselvan
authors.forename: Mani
authors.initials: M
authors.lastname: Menon
authors.forename: Sivagnanam
authors.initials: S
authors.lastname: Thamilselvan
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000473:pathology*; D006861:Hydrogen Peroxide / Q000378:metabolism; D007684:Kidney Tubules / Q000473:pathology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D018374:LLC-PK1 Cells; D015227:Lipid Peroxidation / Q000187:drug effects; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000502:physiology*; D010070:Oxalates / Q000494:pharmacology*; D018384:Oxidative Stress / Q000502:physiology*; D051571:Protein Kinase C-alpha / Q000037:antagonists & inhibitors*; D051745:Protein Kinase C-delta / Q000037:antagonists & inhibitors*; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors; D015398:Signal Transduction / Q000502:physiology; D013347:Subcellular Fractions / Q000378:metabolism; D013552:Swine; D014162:Transfection
chemical_list: D004791:Enzyme Inhibitors; D010070:Oxalates; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase; D019255:NADPH Oxidases; D051571:Protein Kinase C-alpha; D051745:Protein Kinase C-delta
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