Sepsis-induced increase in iNOS protein expression in brain tissues.

<p>(A) Time course of changes in cerebral expression of iNOS protein after CLP. β-Actin served as a loading control. The result is representative of two independent experiments. (B) Comparison of iNOS protein expression between sham-operated and CLP septic groups at 24 h after surgery. The summarizing data are presented as iNOS/GAPDH expressed relative to the unoperated control. Mean of data from three animals in each group is presented, with SEM by vertical lines. *<i>p</i><0.05.</p

Effect of edaravone treatment on neuronal degeneration in cerebral cortex after sepsis.

<p>In the vehicle-treated CLP septic group, there was serious neuronal degeneration in cerebral cortex (B) compared with sham control (A). In the edaravone-treated CLP septic group, normal-appearing neuronal findings were obtained in cerebral cortex (C). Tissues were harvested 24 h after surgery. Magnified view is shown in each inset. (D) Estimation of densely-stained cells in cerebral cortex. Counts of densely-stained cells were made in the sections at a final magnification of ×100. Mean of data from eight animals in each group is presented, with SEM by vertical lines. **<i>p</i><0.01 versus control. #<i>p</i><0.05 versus CLP treated with vehicle.</p

Sepsis-induced increase in extravasation of sodium fluorescein (A) and Evans blue (B) in brains.

<p>Tissues were harvested 24 h after surgery. Mean of data from 4–6 animals in each group is presented, with SEM by vertical lines. *<i>p</i><0.05 versus sham-operated control.</p

Morphological damage of neurons in cerebral cortices from septic mice.

<p>Images from hematoxylin and eosin-stained sections represent cerebral cortices from sham-operated control (A) and from CLP septic mice (B). Tissues were harvested 24 h after surgery. Bottom shows high-magnification images. Arrows indicate densely-stained cells. Shown are representative micrographs from three independent experiments in which the same results were obtained.</p

Sepsis-induced changes in NADPH oxidase activity, NADPH oxidase components p47<i>^phox</i> and p67<i>^phox</i> expression, and oxidative stress parameter in brain tissues.

<p>(A) Time course of changes in NADPH oxidase activity. Mean of data from six animals in each group is presented, with SEM by vertical lines. *<i>p</i><0.05 versus control. (B) Western immunoblotting showing up-regulation of p47<i>^phox</i> and p67<i>^phox</i> protein expression after CLP. The result is representative of two independent experiments. (C) The mRNA levels for p47<i>^phox</i> and p67<i>^phox</i> were quantified by real-time PCR. The data are expressed as a fold increase above control (sham operation) normalized GAPDH. Mean of data from six animals in each group is presented, with SEM by vertical lines. *<i>p</i><0.05 versus sham-operated control. (D) Levels of 8-OHdG. Mean of data from ten animals in each group is presented, with SEM by vertical lines. *<i>p</i><0.05 versus control.</p

Morphological damage of neurons in hippocampal areas from septic mice.

<p>Images from hematoxylin and eosin-stained sections represent hippocampal areas from sham-operated control (A) and from CLP septic mice (B). Tissues were harvested 24 h after surgery. Bottom shows high-magnification images. Arrows indicate densely-stained cells. Representative micrographs from two independent experiments are shown.</p

Effect of edaravone treatment on neuronal degeneration in hippocampus after sepsis.

<p>In the vehicle-treated CLP septic group, there was serious neuronal degeneration in hippocampus (B) compared with sham control (A). In the edaravone-treated CLP septic group, normal-appearing neuronal findings were obtained in hippocampus (C). Tissues were harvested 24 h after surgery. Magnified view is shown in each inset. Shown are representative micrographs from two independent experiments in which the same results were obtained.</p

Sepsis-induced up-regulation of proinflammatory cytokine expression in brain and lung tissues.

<p>Tissues were harvested 12 and 24 h after CLP. In (A) and (C), the mRNA levels for TNF-α, IL-1β, and IL-6 were quantified by real-time PCR. The data are expressed as a fold increase above control (sham operation) normalized GAPDH. Mean of data from 3–5 animals in each group is presented, with SEM by vertical lines. *<i>p</i><0.05, **<i>p</i><0.01 versus sham-operated control. (B) Western blots of TNF-α, IL-1β, and IL-6. GAPDH served as loading control. Representative images from two separate experiments are shown.</p