SE-induced vasogenic edema formation via ETB receptor-mediated NADPH oxidase pathway.

<p>(<b>A</b>–<b>E</b>) Effects of BQ788 and apocynin on SE-induced up-regulation of 4-HNE immunoreactivity in astrocytes 12 h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE induced animals; #P < 0.05 versus vehicle-treated animals; one-way ANOVA followed by Tukey’s test. (<b>F</b>–<b>J</b>) Effects of BQ788 and apocynin on dystrophin and AQP4 expression 12h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE induced animals; #P < 0.05 versus vehicle-treated animals; one-way ANOVA followed by Tukey’s test. (<b>K</b>–<b>M</b>) Effect of apocynin on dystrophin and AQP4 mRNA/protein expression levels after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE animals; #P < 0.05 versus vehicle-treated animals; one-way ANOVA followed by Tukey’s test. (<b>N</b>–<b>Q</b>) Quantification of the attenuation of vasogenic edema formation by BQ788 and apocynin in the PC (means ± s.e.m., n = 5, respectively); *P < 0.05 versus vehicle treated animals; #P < 0.05 versus BQ788-treated animals; one-way ANOVA followed by Tukey’s test. Scale bars: <b>A-D</b>, 12.5 μm; <b>insertion</b> in <b>B</b>, 10 μm; <b>F–I</b>, 25 μm; <b>N–P</b>, 400 μm.</p

SE-induced vasogenic edema formation via the ETB receptor-mediated eNOS pathway.

<p>(<b>A</b>–<b>B</b>) Effect of BQ788 on SMI-71 expression and ETB receptor expression 12 h after SE. (<b>C</b>) Effect of BQ788 on ET_B receptor mRNA expression 12 h after SE (means ± s.e.m., n = 5, respectively); paired Student’s t-test. (<b>D</b>) Nitrate/nitrite (NO products) concentration in the PC after SE (mean ± s.d., n = 5): *P < 0.05 versus basal level; paired Student’s t-test. (<b>E</b>–<b>G</b>) Effects of BQ788 and Cav1-peptide on eNOS mRNA/protein expression 12 h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE animals; #P < 0.05 versus vehicle-treated animals; P < 0.05 versus BQ788-treated animals; one-way ANOVA followed by Tukey’s test. (H–J) Effects of of BQ788 and Cav1-peptide on eNOS, SMI-71 and NT expression 12 h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE animals; #P < 0.05 versus vehicle-treated animals; P < 0.05 versus BQ788-treated animals; one-way ANOVA followed by Tukey’s test. (<b>K</b>–<b>N</b>) Quantification of vasogenic edema formation 3days after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus vehicle-treated animals; #P < 0.05 versus BQ788-treated animals; one-way ANOVA followed by Tukey’s test. Scale bars: <b>A, B, H</b> and <b>I</b>, 25 μm; <b>K-M</b>, 400 μm.</p

Scheme depicting the role of the ET-1 in vasogenic edema formation induced by SE.

<p>Scheme depicting the role of the ET-1 in vasogenic edema formation induced by SE.</p

TNF-α/NFκB-mediated ET-1 release and expression in the PC following SE.

<p>(<b>A</b>) The extracellular ET-1 concentration in the PC after SE (mean ± s.d., n = 5): *P < 0.05 versus basal level; paired Student’s t-test. (<b>B</b>) The effect of sTNFp55R, and SN50 pretreatment on ET-1 mRNA expression 12 h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE animals; #P < 0.05 versus vehicle-treated animals; one-way ANOVA followed by Tukey’s test. (<b>C</b>–<b>F</b>) ET-1 expression in neurons and endothelial cells 12 h after SE. (<b>G</b>–<b>H</b>) Effects of sTNFp55R and SN50 pretreatment on ET-1 expression and SMI-71 immunoreactivity 12 h after SE. (<b>I</b>–<b>K</b>) Quantification of ET_B receptor levels by western blotting and qRT-PCR in the PC 12 h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE animals; #P < 0.05 versus vehicle-treated animals; one-way ANOVA followed by Tukey’s test. (<b>L</b>–<b>O</b>) Effect of SN50 on ETB receptor expression and SMI-71 immunoreactivity 12h after SE. Scale bars: <b>C–H</b>, <b>L-O</b>, 25 μm.</p

ETB receptor-mediated reduction of dystrophin and AQP4 expression in astrocytes.

<p>(<b>A</b>–<b>C</b>) Effects of BQ788 and Cav1-peptide on dystrophin and AQP4 mRNA/protein expression levels 12 h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE animals; #P < 0.05 versus vehicle-treated animals; P < 0.05 versus BQ788-treated animals; one-way ANOVA followed by Tukey’s test. (D–F) Effects of BQ788 and Cav-1 peptide on dystrophin and AQP4 expression in astrocytes 12 h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE animals; #P < 0.05 versus vehicle-treated animals; P < 0.05 versus BQ788-treated animals; one-way ANOVA followed by Tukey’s test. Scale bar: <b>D</b> and <b>E</b>, 12.5 μm.</p

The roles of TNF-α in SE-induced vasogenic edema in the PC.

<p>(<b>A</b>) The extracellular TNF-α concentration after SE (mean ± s.d., n = 5): *P < 0.05 versus the basal level; paired Student’s t-test. (<b>B</b> and <b>C</b>) Quantification of western blots for TNF-α protein expression and p65-Thr435 NFκB phosphorylation 12 h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 by Student’s t-test. (<b>D</b>–<b>I</b>) Immunofluorescence data for TNFp75R, p65-Thr435 NFκB phosphorylation and SMI-71 12 h after SE. (<b>J</b>–<b>K</b>) Effects of TNFp55R and SN50 on p65-Thr435 NF-κB phosphorylation and SMI-71 immunoreactivity. (<b>L</b>–<b>M</b>) Quantification of the fluorescence intensities of SMI-71 expression and p65-Thr435 NFκB phosphorylation 12 h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE animals; #P < 0.05 versus vehicle-treated animals; one-way analysis of variance (ANOVA) followed by Tukey’s test. (<b>N</b>–<b>Q</b>) Quantification of vasogenic edema attenuation by sTNFp55R and SN50 3 days after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus vehicle treated animals by one-way ANOVA followed by Tukey’s test. Scale bars: <b>D–K</b>, 25 μm; <b>O-Q</b>, 400 μm.</p

ETB receptor-mediated p47phox expression in astrocytes.

<p>(<b>A</b>–<b>C</b>) Effects of BQ788 and Cav1-peptide on p47phox mRNA/protein expression level 12 h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE animals; #P < 0.05 versus vehicle-treated animals; P < 0.05 versus BQ788-treated animals; one-way ANOVA followed by Tukey’s test. (D–E) Effects of BQ788 and Cav-1 peptide on p47phox expression in astrocytes 12 h after SE (means ± s.e.m., n = 5, respectively); *P < 0.05 versus non-SE animals; #P < 0.05 versus vehicle-treated animals; P < 0.05 versus BQ788-treated animals; one-way ANOVA followed by Tukey’s test (<b>E</b>). Scale bar: <b>D</b>, 25 μm.</p