Phenylephrine-mediated contraction in mesenteric arteries submitted to high blood flow.

<p>Phenylephrine-mediated contraction was measured in mesenteric arteries submitted to a chronic increase in blood flow (high flow: HF, right panel) and in control arteries submitted to normal flow (NF, left panel). Arteries were isolated from ovariectomized rats treated with resveratrol 5.0 (R5.0 or Resv. 5.0, n = 9 rats) or 37.5mg/kg (R37.5 or Resv. 37.5, n = 9 rats) or with the vehicle (n = 10 rats). Mean ± sem is represented. *P<0.05, HF versus NF arteries. ^#P<0.05, R37.5 versus vehicle.</p

Arterial diameter in mesenteric arteries submitted to high blood flow.

<p>Luminal diameter was measured in mesenteric arteries submitted to a chronic increase in blood flow (high flow: HF) and in control arteries submitted to normal flow (NF). Arteries were isolated from ovariectomized rats treated with resveratrol 5.0 (B, n = 9 rats) or 37.5mg/kg (C, n = 9 rats) or with the vehicle (A, n = 10 rats). In a separate series of experiments, ERalpha-/- mice were ovariectomized and treated with the solvent or with resveratrol 5mg/kg (D, n = 4 mice per group). Mean ± sem is represented. *P<0.05, HF versus NF arteries.</p

Mitochondrial proteins expression level.

<p>The expression level of of Cytochrome-C Oxidase IV (COX IV, A), sirtuin-1 (B), pGC1alpha (C), and Cytochrome C (D) was determined using Western-blot analysis in mesenteric arteries (left panel). The ratio of the expression level in HF to the expression level in NF arteries is shown in the right panel. Arteries were isolated from ovariectomized rats treated with resveratrol 5.0 (Resv. 5.0, n = 9 rats) or 37.5mg/kg (Resv. 37.5, n = 9 rats) or with the vehicle (n = 10 rats). Mean ± sem is represented (n = 12 rats per group). Blots are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146148#pone.0146148.s003" target="_blank">S3A–S3D Fig</a>. *P<0.05, effect of the treatment: Resv. 5.0 or Resv. 37.5 versus vehicle.</p

Expression level of eNOS (A), p67 phox (B), Gp91 phox (C) and ERK1/2 activation (D, ratio of phospho-ERK1/2 to ERK1/2) expression level.

<p>The ratio of protein expression in HF to the expression level in NF arteries is shown in the right panel. Arteries were isolated from ovariectomized rats treated with resveratrol 5.0 (Resv. 5.0, n = 9 rats) or 37.5mg/kg (Resv. 37.5, n = 9 rats) or with the vehicle (n = 10 rats). Mean ± sem is represented. Blots are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146148#pone.0146148.s003" target="_blank">S3A–S3D Fig</a>. *P<0.05, effect of the treatment: Resv. 5.0 or Resv. 37.5 versus vehicle.</p

Arterial structure in mesenteric arteries submitted to high blood flow.

<p>Arterial media thickness (A), media cross-section area (B) and media to lumen ratio (C) were measured in mesenteric arteries submitted to a chronic increase in blood flow (high flow: HF) and in control arteries submitted to normal flow (NF). Arteries were isolated from ovariectomized rats treated with resveratrol 5.0 (R5.0 or Resv. 5.0, n = 9 rats) or 37.5mg/kg (R37.5 or Resv. 37.5, n = 9 rats) or with the vehicle (Veh., n = 10 rats). Mean ± sem is represented. *P<0.05, HF versus NF arteries. ^#P<0.05, R37.5 versus vehicle.</p

Low- and high-collision energy spectra of ions specific to resveratrol-glucuronide, <i>m/z</i> 403.102.

<p>Low- and high-collision energy spectra of ions specific to resveratrol-glucuronide, <i>m/z</i> 403.102.</p

Concentrations of <i>trans</i>-resveratrol and its hydrophilic metabolites and hydrophilic metabolites of dihydro-resveratrol (DHR).

<p>Mean values in µmol.L⁻¹. Values expressed as “<i>trans</i>-resveratrol-4′-O-ß-glucuronide equivalent” for <i>trans</i>-resveratrol glucuronide sulfate and DHR-glucuronide sulfate; as “<i>trans</i>-resveratrol-3-O-ß-glucuronide equivalent” for the DHR-glucuronide and as “<i>trans</i>-resveratrol-3-sulfate equivalent” for the DHR-sulfate. CV% between mouse lemurs (round brackets), range [square brackets]. There were three determinations for each concentration. <i>n</i>: sample size.</p><p>LLOQ: lower limit of quantification.</p

Chemical structure of A: <i>Trans</i>-resveratrol; B: <i>trans</i>-resveratrol-¹³C₆; C: <i>trans</i>-resveratrol-3-sulfate (R1 = R2 = R3 = R4 = H) or <i>trans</i>-resveratrol-3-sulfate-D4 (R1 = R2 = R3 = R4 = D); D: <i>trans</i>-resveratrol-3-O-ß-D-glucuronide (R1 = R2 = R3 = R4 = H) or <i>trans</i>-resveratrol-3-O-ß-D-glucuronide-D₄ (R1 = R2 = R3 = R4 = D); E: <i>trans</i>-resveratrol-4′-O-ß-D-glucuronide (R1 = R2 = R3 = R4 = H) or <i>trans</i>-resveratrol-4′-O-ß-D-glucuronide-D₄ (R1 = R2 = R3 = R4 = D).

<p>Chemical structure of A: <i>Trans</i>-resveratrol; B: <i>trans</i>-resveratrol-¹³C₆; C: <i>trans</i>-resveratrol-3-sulfate (R1 = R2 = R3 = R4 = H) or <i>trans</i>-resveratrol-3-sulfate-D4 (R1 = R2 = R3 = R4 = D); D: <i>trans</i>-resveratrol-3-O-ß-D-glucuronide (R1 = R2 = R3 = R4 = H) or <i>trans</i>-resveratrol-3-O-ß-D-glucuronide-D₄ (R1 = R2 = R3 = R4 = D); E: <i>trans</i>-resveratrol-4′-O-ß-D-glucuronide (R1 = R2 = R3 = R4 = H) or <i>trans</i>-resveratrol-4′-O-ß-D-glucuronide-D₄ (R1 = R2 = R3 = R4 = D).</p

Chromatograms: variations in the intensity of <i>m/z</i> 227.07 (a) and <i>m/z</i> 229.087 (b) vs. time.

<p><b>a</b>: Time 2 h after intake of resveratrol. High-collision energy spectrum at 2.05 min. <b>b</b>: Time 24 h after intake of resveratrol. High-collision energy spectrum at 2.11 min.</p

<i>m/z</i> average and accuracy (ppm) for precursor and daughter ions of resveratrol and their metabolites.

<p>RGS = <i>trans</i>-resveratrol-glucuronide-sulfate; 4′RG = <i>trans</i>-resveratrol-4′-O-ß-glucuronide; 3RG = <i>trans</i>-resveratrol-3-O-ß-glucuronide; RS = <i>trans</i>-resveratrol-sulfate, R = resveratrol; DHRGS = DHR-glucuronide-sulfate; DHRG = DHR-glucuronide; DHRS = DHR-sulfate.</p><p>Standard samples: 3×5 injections; plasma samples: 17×2 injections.</p><p><i>n</i>: Number of injections where an ion was found.</p