engleski

In Vitro Studies of Oxidative Stress in the Kidney Stone Formation

Urolithiasis is a result of formation and retention of crystals within the kidneys. Numerous causes may lead to kidney stone formation. However, most kidney stones are predominantly composed of calcium oxalate which predominantly generates free radicals that are toxic to the renal tubular cells. The aim was to study the toxic effect of oxalate to renal cells and possible inhibition of oxalate induced oxidative stress by antioxidant treatment. Pig kidney epithelial cells (LLC-PK1) were used as in vitro model of urolithiasis. Oxidative stress was induced by exposure of LLC-PK1 cells to sodium oxalate (NaOX). In order to prevent oxidative stress LLC-PK1 cells were pretreated with different concentrations of vitamin E, an antioxidant. The cytotoxicity of oxalate against LLC-PK1 cells was determined by cell counting and by light microscopy. The oxidative stress was evaluated by expression of superoxide dismutase (SOD), which was done by RT-PCR and immunohistochemically (using anti-SOD I antibody In the cells treated with NaOX only, necrosis was observed proportionally to the concentration of NaOX. Interestingly, cells treated with vitamin E prior NaOX exposure showed lover levels of necrosis by light microscopy as well as by cell counting. Positive correlation of SOD expression was observed in all groups of cells. An antioxidant pretreatment with vitamin E of the cells later exposed to oxalate toxicity is able to hamper oxalate induced oxidative stress generation in kidney cells. Vitamin E may play role in prevention of urolithiasis. Further evaluation of its potential as a prevention agent of urolithiasis are needed.

urolithiasis; antioxidantt; prevention; LLC-PK1; vitamin E

Urolithiasis is a result of formation and retention of crystals within the kidneys. Numerous causes may lead to kidney stone formation. However, most kidney stones are predominantly composed of calcium oxalate which predominantly generates free radicals that are toxic to the renal tubular cells. The aim was to study the toxic effect of oxalate to renal cells and possible inhibition of oxalate induced oxidative stress by antioxidant treatment. Pig kidney epithelial cells (LLC-PK1) were used as in vitro model of urolithiasis. Oxidative stress was induced by exposure of LLC-PK1 cells to sodium oxalate (NaOX). In order to prevent oxidative stress LLC-PK1 cells were pretreated with different concentrations of vitamin E, an antioxidant. The cytotoxicity of oxalate against LLC-PK1 cells was determined by cell counting and by light microscopy. The oxidative stress was evaluated by expression of superoxide dismutase (SOD), which was done by RT-PCR and immunohistochemically (using anti-SOD I antibody In the cells treated with NaOX only, necrosis was observed proportionally to the concentration of NaOX. Interestingly, cells treated with vitamin E prior NaOX exposure showed lover levels of nec