ECTS2016 Poster Presentations Other diseases of bone and mineral metabolism (52 abstracts)
In vitro model of antioxidant prevention of urolithiasis
Tomislav Kizivat , Martina Smolic , Ines Bilic-Curcic , Robert Smolic , Ivana Maric , Hrvoje Roguljic , Maja Tolusic-Levak , Lucija Kuna , Aleksandar Vcev & Antun Tucak
University of Osijek, Faculty of Medicine, Osijek, Croatia.
Urolithiasis is characterized by formation and retention of solid crystals within the urinary tract. There are numerous causes that may lead to urinary stone formation. However, kidney stones are mostly composed of calcium oxalate that predominantly generates free radicals that are toxic to renal tubular cells.
The aim of the study is to explore the toxic effect of oxalate to renal epithelial cells and to explore possible effects of antioxidants on its inhibition.
Two cell lines were used as in vitro model of urolithiasis: Madin Darby canine kidney cells subtype II (MDCKII) and pig kidney epithelial cells (LLC-PK1). Oxidative stress was induced by exposure of cells to sodium oxalate (NaOX) in concentration of 8 mM. In order to prevent oxidative stress, cells were treated with three different concentrations (0.5, 0.1 and 0.05 ng/ml) of L-arginine, an antioxidant. Cytotoxicity of NaOX and the effects of L-arginine were determined by cell counting and light microscopy. The oxidative stress was evaluated by expression of superoxide dismutase (SOD), immunohistochemically (with antiSODI antibody) and by RT-PCR.
In both cell lines treated with NaOX only, cell necrosis was observed. Cell survival in MDCKII cell line was around 60%, while in LLC-PK1 cell line it was significantly less. In both cell lines, cells pretreated with L-arginine prior to NaOX exposure showed lower levels of necrosis, and significantly higher levels of cell survival. Positive correlation of SOD expression was observed in all groups of cells, by immunohistochemistry and by RT-PCR.
Our results indicate that an antioxidant pretreatment with L-arginine of cells later exposed to oxalate toxicity is able to hamper oxalate-induced oxidative stress in kidney epithelial cells and as such could play a role in prevention of urolithiasis. More studies to further evaluate its potential as a prevention agent of urolithiasis are obligatory.
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