Oxidative Stress Markers in Kidney

ROS level in kidney was assayed as previously described [18 (link)]. The nonpolar compound dihydrodichlorofluorescein diacetate (H2 DCFH-DA), after conversion to a polar derivative by intracellular esterases, can rapidly react with ROS to form the highly fluorescent compound dichlorofluorescein. The commercial available assay kit (E004, Nanjing Jiancheng Bioengineering Institute, Jiangsu, China) was used. As previously described [19 (link)], malondialdehyde (MDA) and superoxide dismutase (SOD) were detected in the kidney tissues. The content of MDA was determined by the thiobarbituric acid method (A003-1, Nanjing Jiancheng Bioengineering Institute, Jiangsu, China). The SOD activity assay kit (A001-1, Nanjing Jiancheng Bioengineering Institute, Jiangsu, China) was enrolled. SOD activity was determined by inhibition of nitroblue tetrazolium reduction due to superoxide anion generation by a xanthine-xanthine oxidase system. All the assays were carried out according to the manufacture's protocol using spectra microplate reader (model A-5082, Tecan, Australia).