| Urolithiasis is a common urinary tract disorder of dogs and cats. The incidence of canine calcium oxalate uroliths has increased over the past 20 years, with a reciprocal decrease in the proportion of struvite uroliths. The reasons for the increase in submissions of calcium oxalate from dogs are likely multifactorial and could include breed and nutritional changes. With the development of the study of human calcium oxalate stone formation, numberous of theory and practice appeared. The experiment was designed according to the theory being broadly approved in calcium oxalate stone formation which was that stone formation requires mineral nucleation, growth, aggregate, mineral and renal tubular epithelial cell adhesion.The aim of this study is to investigate the role of calcium oxalate on renal tubular epithelial cells during the process of calcium oxalate stone formation.In this research, Madin-Darby canine kidney cells(MDCK-cell)were cultured in vitro and exposed to different concentrations of calcium oxalate(Ca Ox, 300μg/m L,600μg/m L,900μg/m L) for 1h、2h、6h、12h、24h and 48 h. After specified periods of exposure to Ca Ox, the status of the cell and cell morphologyin and the situation of Ca Ox crystal conglutination in petri dish were observed with microscope. Treat the cell sample using standard ultrasonic cell disintegration procedures and the cell viability was measured by CCK-8 kits. The content of cellular protein, Hydrogen peroxide, malonaldehyde, lactate dehydrogenase, and the vitality of GGT and superoxide dismutase were determined by kits for detect cellular oxidative damage. Finally, collect all the data and analyze the relation among cell viability, Ca Ox crystal conglutination and cellular oxidative damage.The result showed that exposure to the same concentration of Ca Ox crystals, with time go on, the oxidative damage become serious the viability of cell and SOD activity reduced and more Ca OX crystal conglutinated on MDCK cell. At the same time point, MDCK-cell exposed to higher concentration of Ca Ox suffered worse oxidative damage, crystal conglutination, as well as a reduction of cell viability and SOD activity.Results indicate that calcium oxalate has cytotoxicosis effect on MDCK cell, include oxidative damage by ROS. During this process, calcium oxalate crystal conglutination starts and become more fast through cell debris that produced by cell injured. Therefore, higher concentration of Caox in canine urine and longer existing time lead to more seriously injury to canine kidney and lead to more crystal conglutination that promote canine calcium oxalate uroliths. |
