title: Selective Rac1 inhibition protects renal tubular epithelial cells from oxalate-induced NADPH oxidase-mediated oxidative cell injury.
abstract: Oxalate-induced oxidative cell injury is one of the major mechanisms implicated in calcium oxalate nucleation, aggregation and growth of kidney stones. We previously demonstrated that oxalate-induced NADPH oxidase-derived free radicals play a significant role in renal injury. Since NADPH oxidase activation requires several regulatory proteins, the primary goal of this study was to characterize the role of Rac GTPase in oxalate-induced NADPH oxidase-mediated oxidative injury in renal epithelial cells. Our results show that oxalate significantly increased membrane translocation of Rac1 and NADPH oxidase activity of renal epithelial cells in a time-dependent manner. We found that NSC23766, a selective inhibitor of Rac1, blocked oxalate-induced membrane translocation of Rac1 and NADPH oxidase activity. In the absence of Rac1 inhibitor, oxalate exposure significantly increased hydrogen peroxide formation and LDH release in renal epithelial cells. In contrast, Rac1 inhibitor pretreatment, significantly decreased oxalate-induced hydrogen peroxide production and LDH release. Furthermore, PKC α and δ inhibitor, oxalate exposure did not increase Rac1 protein translocation, suggesting that PKC resides upstream from Rac1 in the pathway that regulates NADPH oxidase. In conclusion, our data demonstrate for the first time that Rac1-dependent activation of NADPH oxidase might be a crucial mechanism responsible for oxalate-induced oxidative renal cell injury. These findings suggest that Rac1 signaling plays a key role in oxalate-induced renal injury, and may serve as a potential therapeutic target to prevent calcium oxalate crystal deposition in stone formers and reduce recurrence.
authors.affiliation: Vattikuti Urology Institute, Suite 2D-34, Henry Ford Health System, One Ford Place, Detroit, MI 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: D000634:Aminoquinolines / Q000494:pharmacology; D000818:Animals; D019610:Cytoprotection; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D007684:Kidney Tubules / Q000187:drug effects* / Q000378:metabolism; D018374:LLC-PK1 Cells; D019255:NADPH Oxidases / Q000502:physiology*; D010070:Oxalates / Q000633:toxicity*; D018384:Oxidative Stress; D011493:Protein Kinase C / Q000502:physiology; D021381:Protein Transport / Q000187:drug effects; D011743:Pyrimidines / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction; D013552:Swine; D020830:rac1 GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000502:physiology*
chemical_list: D000634:Aminoquinolines; C487513:NSC 23766; D010070:Oxalates; D011743:Pyrimidines; D017382:Reactive Oxygen Species; D019255:NADPH Oxidases; D011493:Protein Kinase C; D020830:rac1 GTP-Binding Protein