Myogenic pressure curves of middle cerebral arteries from WKY (n = 9, open squares), SHR (n = 6, full squares) and SHR treated with telmisartan (SHR + TELMI, 2 mg/kg per day, n = 7, open triangles).

<p>Myogenic pressure curves of middle cerebral arteries from WKY (n = 9, open squares), SHR (n = 6, full squares) and SHR treated with telmisartan (SHR + TELMI, 2 mg/kg per day, n = 7, open triangles).</p

Impact of free GSNO on MAP, (panel A), HR, (panel B) and PAP (panel C).

<p>p_interaction, p_dose and p_time < 10⁻⁴ for all parameters (two-way ANOVA; post-hoc Bonferroni test). Each animal (n = 7) received all the treatments. Arrows indicate for each dose the time span where significant differences <i>vs</i>. PBS were observed. Grey boxes represent the duration of anesthesia and red vertical dashed lines the time of injection.</p

Impact of the treatments on whole blood platelet aggregation (aggregometry AUC).

<p>Treatments were: PBS (n = 6), empty microparticles (n = 8), free GSNO at 7.5mg/k g(n = 5) or 30 mg/kg (n = 5), <i>in situ</i> implants loaded with 7.5 ± 0.8 mg/kg (n = 7) or 32.1 ± 4.3 mg/kg (n = 10) of GSNO and <i>in situ</i> microparticles loaded with 7.9 ± 0.6 mg/kg (n = 7) or 31.4 ± 3.6 mg/kg (n = 10) of GSNO. The red vertical dashed lines represent the time of injection. †: p < 0.05 <i>vs</i>. 1 h following substance administration within the same group (one-way ANOVA; post-hoc Bonferroni test).</p

Impact of formulated GSNO at 30 mg/kg on MAP (panel A), HR (panel B) and PAP (panel C).

<p>Each animal (n = 7) received all the treatments (Implants: 31.9 ± 0.5; Microparticles 30.9 ± 0.6 mg/kg). Day-night cycles are marked in days (D1, D2 and D3) and nights (N1, N2) periods. For more clarity, after the first hour, only one point for each hour was represented. *: p < 0.05 <i>vs</i>. PBS (one-way ANOVA; post-hoc Bonferroni test).</p

<i>In vitro</i> releases of GSNO from <i>in situ</i> forming implants and microparticles.

<p>GSNO loadings were 1.25% or 5% m/m (corresponding respectively to 7.5 mg/kg and 30 mg/kg of GSNO for <i>in vivo</i> experiments) based on the amount of polymer/GSNO/solvent solution. Panel A represents the release of GSNO expressed as % of the initial load; Panel B represents the release of GSNO in absolute values. Values are mean ± s.e.m. (n = 3).</p

Detailed impact of formulated GSNO at 7.5 mg/kg on MAP (panel A), HR (panel B) and PAP (panel C) within the first half day following administration.

<p>Each animal (n = 7) received all the treatments (Implants: 7.4 ± 0.2; Microparticles: 7.7 ± 0.2 mg/kg).</p

Impact of the treatments on neurological score, infarct size and edema volume.

<p>Treatments were: empty microparticles (n = 8), free GSNO at 7.5 mg/kg (n = 8) and <i>in situ</i> implants loaded at 30.1 ± 1.5 mg/kg (n = 8) or microparticles at 31.4 ± 0.9 mg/kg (n = 6) of GSNO, injected in male Wistar rats 2 h after thromboembolic stroke induction. Neurological score (panel A, maximal score 160) was measured 24 and 48 h after stroke induction, infarct size (panel B) and edema volume (panel C) 48 h after. *: p < 0.05 <i>vs</i>. empty microparticles; #: p < 0.05 <i>vs</i>. free GSNO at 7.5 mg/kg; <i>ο</i>: p < 0.05 <i>vs</i>. <i>in situ</i> implants (Kruskal-Wallis; post-hoc Mann-Whitney test).</p

Effects of modulation of GGT activity on GSNO-mediated ^•NO production in aortic tissue.

<p>Panel A: ^•NO production from rat aortic rings exposed to GSNO (1 mM) or GSNO (1 mM) in the presence of SBC (20 mM), glycylglycine (20 mM) or L-NAME (300 µM). Data are means ± SEM of 4–7 experiments, * p<0.05 <i>versus</i> GSNO. Panels B, C, D: representative fluorescence micrographs (×20) of 10-µm sections of rat aortic rings loaded for 2 h with 4,5-diaminofluorescein diacetate (DAF-2 DA, 10 µM) showing ^•NO production following 15-min incubation with GSNO (1 mM, panel C) or GSNO in the presence of SBC (20 mM, panel D) or glycylglycine (20 mM, panel B). Scale bar, 50 µm. All pictures were captured under constant exposure time, gain and offset.</p

Histochemical localization of GGT activity in rat aortic tissue.

<p>Typical micrographs (×16) corresponding to histochemical visualization of γ-glutamyltransferase (GGT) activity in endothelium-intact rat aortic rings prepared in the presence of γ-glutamyl-4-methoxy-2-naphthylamide (GMNA, panel A) or in the presence of GMNA and SBC (20 mM, panel B) and in endothelium-denuded aortic rings prepared in the presence of GMNA (panel C). Scale bar, 100 µm.</p

Hypotension-induced changes in pial arteriolar diameters.

<p>Internal diameters of pial arterioles (ID, µm) during decrease in systemic mean arterial blood pressure (mmHg), <b>A</b>: in 4–5 month-old normotensive rats (WKY, empty circles) and hypertensive vehicle-treated rats (SHR, full triangles); and <b>B</b>: in hypertensive rats treated for 10 days with telmisartan (TELMI, 2 mg/kg per day, empty squares) or candesartan cilexetil (CANDE, 10 mg/kg per day, full squares). n = 4–8 per group, m±sem.</p