Preventive effects of L-arginine on oxidative damage and crystallization in cellular model of urolithiasis of distal renal tubules

Cilj istraživanja: Odrediti toksični učinak natrijevog oksalata u različitim koncentracijama i u različitom vremenskom periodu izloženosti na epitelnim stanicama distalnih bubrežnih tubula (MDCK1) te odrediti učinak antioksidansa L-arginina pri različitim koncentracijama na proces toksičnosti oksalata i kristalizacije i izražaj mRNA superoksid dismutaze u staničnom modelu oksalatne urolitijaze distalnih bubrežnih tubula (MDCK1). Nacrt studije: Laboratorijska znanstvena studija. Materijali i metode: Korištena stanična kultura je Madin-Darby, a dobivena je od psećih bubrežnih stanica podtipa I (MDCK I). Za ispitivanje toksičnosti, korišten je natrijev oksalat (pročišćen do ≥ 99 %) koji je inkubiran sa staničnom kulturom te se vijabilnost stanica nakon inkubacije pregledala bojenjem bojom „trypan blue“. Za ispitivanje zaštitnog djelovanja protiv oksalatne toksičnosti korišten je L-arginin kupljen od Sigma Chemicalsa (St. Louis, MO) koji se inkubirao s korištenim stanicama u zasebnoj seriji zajedno s natrijevim oksalatom. Rezultati: Nakon inkubiranja stanica s različitim koncentracijama natrijevog oksalata, pokazano je kako natrijev oksalat uzrokuje značajno smanjenje vijabilnosti stanica proporcionalno s povećanjem njegove koncentracije. Jednako tako, otkriveno je kako su stanice inkubirane dodatno i s L-argininom pokazale značajno manje smanjenje vijabilnosti u usporedbi sa stanicama inkubiranima isključivo s natrijevim oksalatom. Zaključak: L-arginin pokazuje uspješno zaštitno djelovanje na stanice distalnih bubrežnih tubula izloženima toksičnom djelovanju natrijevog oksalata te predstavlja potencijalno korisnu terapiju u liječenju urolitijaze.Objectives: Determining the toxic effect of various concentrations of sodium oxalate in various time periods while exposing epithelial cells of distal kidney tubule cells (MDCK 1) to it and determining the effect of the L-arginine antioxidanse on the chrystalisation and toxicity of the sodium oxalate, as well as determining the mRNA expression of superoxide dismutase within our cell model of oxalate urolithiasis of distal kidney tubules (MDCK I). Study design: The study was made in the form of a laboratory scientific research. Materials and methods: The used cell culture was Madin-Darby, which was obtained from the subtype I canine kidney cells (MDCK I). For the purpose of toxicity testing, we used sodium oxalate (≥99% purified) which was incubated with our cell culture. The viability of our cells after the incubation was observed by dying it with the trypan blue dye. For the purpose of testing possible protective effects, we used L-arginine obtained from Sigma Chemicals (St. Louis, MO) which we incubated in separate testing series along with both our cells and sodium oxalate. Results: After incubating our cells with various concentrations of sodium oxalate, it has been observed that sodium oxalate does indeed cause a significant reduction in the viability of our cells, which was proportionate to the concentration of sodium oxalate in use. Also, it was observed that cells incubated with both sodium oxalate and L-arginine displayed a significant reduction in the viability drop, compared to the cells incubated solely with the sodium oxalate. Conclusion: L-arginin displayed a successful protective effect on distal tubule kidney cells which are under the toxic influence of the sodium oxalate and is thus presented as a potentially useful therapy in the treatment of urolithiasis