Measurement of COX-2 expression level in mesenteric resistance arteries isolated from young and old LZ and ZDF rats using immunolabelling (A) and Western-blot analysis (B).

<p>Mean ± SEM is presented (n = 10 per group). #P<0.01, ZDF versus LZ. *P<0.01, old versus young rats.</p

Rat body weight, glycaemia and mean arterial blood pressure (MAP) were measured in 3- and 12-month old LZ and ZDF rats.

<p>Mean±SEM is presented (n = 12 rats per group).</p>*<p>P<0.05, ZDF versus LZ of the same age.</p>#<p>P<0.05, effect of age within each group.</p

Effect of an acute treatment of mesenteric resistance arteries with the COX-2 inhibitor NS398 or NS398 and the antioxidant tempol on acetylcholine-induced relaxation in young (A) and old (B) LZ and ZDF rats.

<p>Mean ± SEM is presented (n = 10 per group). ^#P<0.01, ZDF versus LZ. *P<0.01, old versus young rats. ^&P<0.05, effect of NS398 alone on acetylcholine-induced relaxation. ^$P<0.05, effect of NS398+tempol on acetylcholine-induced relaxation.</p

Measurement of the reactive oxygen species level in mesenteric resistance arteries isolated from LZ and ZDF rats using dihydroethydin microfluorography (A).

<p>The expression level of the NADP(H)-oxidase subunits gp91 and p67 (B) was measured using Western-Blot analysis. Mean ± SEM is presented (n = 10 per group). #P<0.01, ZDF versus LZ. *P<0.01, old versus young rats.</p

Effect of the acute treatment of mesenteric resistance arteries with the antioxydant tempol on acetylcholine-induced relaxation in mesenteric resistance arteries isolated from young (A) and old (B) LZ and ZDF rats.

<p>Mean ± SEM is presented (n = 10 per group). #P<0.01, ZDF versus LZ. *P<0.01, old versus young rats. &P<0.05, effect of tempol on acetylcholine-induced relaxation.</p

Acetylcholine-induced relaxation in young (A) and old (B) LZ and ZDF rats.

<p>Relaxation was obtained before and after NO synthesis inhibition with the NO synthesis blocker L-NAME. The inserts show the precontraction applied before adding acetylcholine. Beta-actin and eNOS expression levels were measured using Western-Blot analysis (typical blots on the right side). Data is given as a ratio eNOS/beta-actin (<b>C</b>). Mean ± SEM is presented (n = 10 per group). #P<0.01, ZDF versus LZ. *P<0.01, old versus young rats.</p

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 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

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