Oxalate-induced activation of PKC- and - regulates NADPH oxidase-mediated oxidative injury in renal tubular epithelial cells

Thamilselvan V, Menon M, Thamilselvan S. Oxalate-induced activation of PKCand regulates NADPH oxidase-mediated oxidative injury in renal tubular epithelial cells. Am J Physiol Renal Physiol 297: F1399–F1410, 2009. First published August 19, 2009; doi:10.1152/ajprenal.00051.2009.—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 PKCand from the cytosol to the cell membrane. Pretreatment of LLC-PK1 cells with specific inhibitors of PKCor 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 PKCand . Silencing of PKCexpression by PKC-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 PKCand -dependent activation of NADPH oxidase is one of the mechanisms responsible for oxalateinduced 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.