Figure 3.
Idh2 gene deletion accelerates H2O2 production, lipid peroxidation, and DNA oxidation in the kidneys after I/R. Idh2+/+ (Idh2 WT) and Idh2−/− (Idh2 KO) mice were subjected to either 25 minutes of bilateral renal ischemia or sham surgery, and then kidneys were harvested 24 hours later. H2O2 (A) and MDA production (B) were determined in the kidney tissue. (C) 4-HNE expression was determined by Western blot analysis using an anti–4-HNE antibody. GAPDH was used as a loading control. (D) The densities of bands were measured using ImageJ software. (E) Kidney sections were subjected to immunohistochemical staining using an 8-OHdG antibody. Images were obtained from the outer medulla. Insert is at high magnification of the dash-lined rectangles. Brown indicates 8-OHdG-positive signal. Number of 8-OHdG–positive cells (F) and intensity of the 8-OHdG stains in the cytosol (G) were determined. Results are expressed as mean±SEM (n=6). *P<0.05 versus respective sham-operated mice; †P<0.05 versus ischemia-operated Idh2 WT mice; #P<0.05 versus sham-operated Idh2 WT mice. Isch, ischemia; KO, knockout; WT, wild type.