Biochemical Analyses of Oxidative Stress Markers in Renal Tissue

The renal tissue was homogenized in Tris-HCl (0.01 M)-EDTA (0.001 M) buffer of pH 7.4 and centrifuged at 12,000× g for 30 min at 4 °C to obtain supernatant for subsequent biochemical assays [6 (link)]. The levels of ROS, NO, H₂O₂, NADPH oxidase, TBARS, carbonylated proteins, GSH, GSSG, SOD, CAT, GPx, GR, and GST in the renal tissue homogenates were measured following the established protocols mentioned earlier. Co-enzymes Q9 and Q10 in the cell lysate were separated and quantified using reverse phase-high performance liquid chromatographic (RP-HPLC) (Dionex, Germany) methods as described by Zhang and co-workers [34 (link)]. DNA fragmentation in the renal cells was measured using the diphenylamine reagent [11 (link)]. The extent of DNA oxidation in the renal cells was measured by quantifying 8-hydroxy-2′-deoxyguanosine (8-OHdG) using a RP-HPLC (Dionex, Idstein, Germany) method [11 (link)].