Intranasal administration of telmisartan decreases amyloid plaques and CD11b staining in the cortex of 3-month-old 5XFAD mice.

<p>Mice were treated with telmisartan (Tel) or with vehicle (N,N-dimethylformamide/polyethylene glycol 400/saline (2:6:2) [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0155823#pone.0155823.ref047" target="_blank">47</a>]) for 3.5 weeks, and their brains were sectioned and immunolabeled with anti-Aβ (red) and anti-CD11b (green) antibodies and countersained with DAPI (blue). <b>(a, c)</b> Representative cortex brain section of WT or 5XFAD mice treated with 1 mg/kg/day telmisartan or with vehicle. Each experiment included 5 mice per group (n = 15 in total). (<b>b, d</b>) Quantification of the average sum of Aβ-stained area <b>(b)</b> and of CD11b-stained area <b>(d)</b>, represented as the mean ± SEM percentage of stained area in the corresponding vehicle-treated group, in at least 3 determinaions. Statistical significance was determined using one-way ANOVA, followed by a Tukey—Kramer Multiple Comparison Test. **P<0.01 vs. WT+Tel; ***P<0.001 vs. WT+Tel, ^^P<0.01 vs. 5XFAD+ vehicle; ^^^P<0.001 vs. 5XFAD + vehicle. <b>(e, f)</b> Representative hippocampal section of 5XFAD mice treated with vehicle. Scale bar is 200 μm.</p

Intranasal administration of telmisartan decreases amyloid plaques and CD11b staining in the cortex and hippocampus of 4-month old 5XFAD mice.

<p>Mice were treated with telmisartan (Tel) or vehicle (N,N-dimethylformamide/polyethylene glycol 400/saline (2:6:2) [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0155823#pone.0155823.ref047" target="_blank">47</a>]) for 8 weeks. The brains of 4-month-old mice were sectioned and immunolabeled with anti-Aβ (red) and anti-CD11b (green) antibodies and countersained with DAPI (blue). (<b>a, c</b>) Representative cortical sections from WT or 5XFAD mice treated with 1 mg/kg/day telmisartan or with vehicle. (<b>e, g</b>) Representative hippocampal sections of WT or 5XFAD mice treated with 1 mg/kg/day telmisartan or with vehicle. Each experiment included 6 mice per group (n = 18 in total). (<b>b, d, f, h</b>) Quantification of the average sum of Aβ-stained area <b>(b, f)</b> or of CD11b-stained area <b>(d, h)</b>, represented as the mean ± SEM percentage of stained area in the corresponding vehicle-treated group in at least 3 determinants. Statistical significance was determined using one-way ANOVA, followed by a Tukey—Kramer Multiple Comparison Test. ***P<0.001 vs. WT+Tel; ^^P<0.01 vs. 5XFAD+vehicle; ^^^P<0.001 vs. 5XFAD+vehicle. Scale bar is 200 μm.</p

Intranasal administration of perindopril decreases amyloid plaques and CD11b staining in the cortex of 3-month old 5XFAD mice.

<p>Mice were treated with perindopril or vehicle (saline) for 3.5 weeks. The brains of 3-month-old mice were sectioned and immunolabeled with anti-Aβ (red) and anti-CD11b (green) antibodies and countersained with DAPI (blue). <b>(a, c)</b> Representative brain section of WT or 5XFAD mice treated with 1 mg/kg/day perindopril or with vehicle. Each experiment included 5 mice per group (n = 15 in total). (<b>b, d)</b> Quantification of the average sum of Aβ-stained area <b>(b)</b> or of CD11b-stained area <b>(d)</b>, are represented as the mean ± SEM percentage of stained area in the corresponding vehicle-treated group in at least 3 determinants. Statistical significance was determined using one-way ANOVA, followed by a Tukey—Kramer Multiple Comparison Test. **P<0.01 vs. WT+perindopril; ***P<0.001 vs. WT+perindopril; ^^^P<0.001 vs. 5XFAD+vehicle. Scale bar is 200 μm.</p

Telmisartan decreased iNOS expression in LPS-induced BV2 microglia.

<p>Cells were incubated with LPS (7 ng/ml) in the presence or absence of telmisartan (Tel), at 1 μM or 5 μM, for 24h. 40 μg protein of whole cell lysate was loaded on 7.5% polyacrylamide-SDS gels. Analysis of iNOS was performed using antibodies against iNOS (130 kDa) and β-actin (40 kDa). Results are representative of two independent experiments and are presented as means ± SEM (overall n = 4–6). ***P<0.001 vs. control; ^^^P<0.001 vs. LPS.</p

Telmisartan decreased NO production in LPS-stimulated BV2 and primary neonatal rat glial cells.

<p>BV2 microglia <b>(a)</b>, primary microglial cells <b>(b)</b> and mixed glial cells <b>(C)</b> were incubated with LPS (7 ng/ml for BV2 cells and 0.5 μg/ml for primary cultures) in the presence or absence of telmisartan (Tel), at 1 μM or 5 μM, for 24h. NO levels were determined in the media and normalized to cells number. <i>Insets</i>: NO levels measured in non-stimulated cells treated with Tel at 1 μM or 5 μM concentrations. Data are presented as means ± SEM and are representatives of 2–3 independent experiments (overall n = 8–12). Statistical significance was determined using one-way ANOVA, followed by a Tukey—Kramer Multiple Comparison Test. ***P < 0.001 vs. control (non-stimulated cells); ^P < 0.05 vs. LPS; ^^^P < 0.001 vs. LPS; ^#P < 0.05 vs. LPS+Telmisartan 1μM; ^###P < 0.001 vs. LPS+Telmisartan 1μM; NS (non-significant) vs. control.</p

Telmisartan attenuated LPS-induced TNF-α and IL1-β release from BV2 microglia cells.

<p>Cells were incubated with LPS (7 ng/ml) in the presence or absence of telmisartan (Tel), at 1 μM or 5 μM, for 24h. Media were collected and analyzed for TNF-α and IL1-β levels and cells were counted. <i>Insets</i>: TNF-α and IL1-β levels measured in non-stimulated cells treated with Tel at 1 μM or 5 μM. Data presented as means ± SEM and are representatives of 2–3 independent experiments (overall n = 8–12). Statistical significance was determined using one-way ANOVA, followed by a Tukey—Kramer Multiple Comparison Test. ***P < 0.001 vs. control (non-stimulated cells); ^^^P < 0.001 vs. LPS; ^###P < 0.001 vs. LPS+Telmisartan 1μM; NS (non-significant) vs. control.</p