Figure 1.

ICH disrupts mitochondrial antioxidative defense and induces oxidative/nitrosative damage in the affected brain hemisphere. A, Immunohistochemistry; immunoreactivity for Mn-SOD (green) in the coronal section of mouse brain at 24 h after ICH. Red dotted line delineates hematoma location (H), and white dotted line delineates ICH-affected area with reduced Mn-SOD expression in the ipsilateral hemisphere. Scale bar, 1000 µm. Cont, Contralateral; Ipsi, ipsilateral. B, Representative Western blot image and illustrating bar graph showing Mn-SOD protein quantitation in the Cont and Ipsi hemispheres at 24 h after ICH. The significance of Mn-SOD/β-actin protein levels was assessed by two-tailed unpaired t test (n = 6 per group), **p < 0.01 (p = 0.0041, Cont vs Ipsi), t value (t = 3.706). C, Mn-SOD mRNA levels in the Cont and Ipsi hemispheres at 24 h after ICH, evaluated by RT-qPCR. Values represent fold change of mRNA levels in ipsilateral hemispheres compared with contralateral hemispheres. Data are shown as mean ± SEM. The significance of Mn-SOD mRNA levels was assessed by two-tailed unpaired t test (n = 8 per group), **p < 0.01 (p = 0.0012, Cont vs Ipsi), t value (t = 4.051). D, In situ detection of intracellular O2^·− in mice at 24 h after ICH that were intravenously injected with 200 µl 1 mg/ml HEt (superoxide indicator) solution 15 min before ICH onset. HEt is incorporated into damaged cells, and on oxidation by superoxide anion is converted to Et, which generates red fluorescence (Murakami et al., 1998; Jung et al., 2009). White dotted line delineates hematoma area (H). Nuclei were counterstained with DAPI (blue). Scale bar, 500 µm. E, Representative Western blot image and quantitative bar graph showing 3-NT protein levels in Cont and Ipsi hemispheres at 3 h after ICH-inducing surgery. The significance of 3-NT/β-actin protein levels was assessed by two-tailed unpaired t test (n = 6 per group), *p < 0.05 (p = 0.0110, Cont vs Ipsi), t value (t = 3.114).