Atorvastatin reduces intracellular and extracellular ROS levels within the atherosclerotic aortic arch.

<p>Female <i>Apoe</i>^<i>-/-</i> mice were fed Western diet to induce advanced atherosclerotic lesions in the aortic arch. Then, mice were treated with vehicle (DMSO) or oral atorvastatin (100 mg/kg per day) for 5 days. <i>(A)</i> ROS levels were analyzed in the aortic arch (red areas). <i>(B–D)</i> Atorvastatin treatment reduced intracellular <i>(B and C)</i> and extracellular ROS levels <i>(D)</i>. <i>(E and F)</i> Plasma cholesterol <i>(E)</i> and triglycerides <i>(F)</i> was reduced by atorvastatin and, to larger extent, by lipid lowering by diet. n = 6 in each group. **p<0.01 vs vehicle, *p<0.05 vs vehicle. One sample t-test (C and D). ANOVA with Dunnet’s test for multiple comparisons (E and F).</p

Intracellular ROS correlates with macrophage content and extracellular ROS with smooth muscle cell content in advanced atherosclerotic lesions Female <i>Apoe</i>^<i>-/-</i> mice were fed Western diet to induce advanced atherosclerotic lesions in the aortic arch.

<p>(<i>A)</i> Intracellular and extracellular ROS were analyzed in the aortic arch (red area) <i>(B)</i> Smooth muscle cell and macrophage content was quantified in the aortic arch by analyzing sections from 4 different levels. <i>(C)</i> Section stained for macrophages (CD68: blue) and smooth muscle cells (α-actin: red). <i>(D and E)</i> Smooth muscle cell content in lesions correlated with extracellular <i>(E)</i> but not intracellular ROS <i>(D)</i>. <i>(F and G)</i> Macrophage content in lesions correlated with intracellular <i>(F)</i> but not extracellular ROS <i>(G)</i>. Linear regression (n = 25). NS (non significant).</p

Atorvastatin does not affect extent of atherosclerosis, lesion cell composition or lesion inflammation Female <i>Apoe</i>^<i>-/-</i> mice were first fed Western diet to induce advanced lesions in the aortic arch.

<p>Then, mice were treated with vehicle (DMSO) or oral atorvastatin (100 mg/kg per day) for 5 days. <i>(A)</i> Atorvastatin did not affect lesion area in the aorta. <i>(B)</i> Atorvastatin did not affect lesion cell composition. <i>(C)</i> Atorvastatin did not affect mRNA levels of inflammatory mediators. n = 6 in each group. Student’s t-test.</p

Intracellular ROS increase before formation of atherosclerotic lesions in hyperlipidemic mice Female <i>Apoe</i>^-/- mice were fed either chow diet or Western diet.

<p><i>(A)</i> The descending thoracic aorta (delineated by dashed line) was relatively spared from atherosclerosis, even in mice fed Western diet. <i>(B)</i> In the descending thoracic aorta, Western diet increased extracellular ROS before formation of atherosclerotic lesions <i>(C)</i> Extracellular ROS levels, in contrast, were increased after formation of atherosclerotic lesions. n = 6–7 in each group. *p>0.05 vs chow diet, One way ANOVA with Dunnett’s multiple comparison test.</p

Lipid lowering by diet does not affect ROS levels within the atherosclerotic aortic arch Female <i>Apoe</i>^<i>-/-</i> mice were first fed Western diet to induce advanced lesions in the aortic arch.

<p>Then, lipids were lowered by switching to chow diet for five days. A control group was maintained on Western diet. <i>(A)</i> Intracellular and extracellular ROS were assessed in the atherosclerotic arch (red). <i>(B)</i> Lipid lowering by diet did not affect intracellular ROS. <i>(C)</i> Lipid lowering by diet did not affect extracellular ROS. NS—non significant. One sample t-test. n = 6 in each group.</p

Patient characteristics (n = 6).

<p>Patient characteristics (n = 6).</p

Severe depletion of ATP and glucose in perinecrotic zone of advanced plaques.

<p>A. Advanced atherosclerotic plaque, delineation in white of viable intima. Extract shows luminal (left) and perinecrotic zone (right) of viable intima. Note high expression of hexokinase II (HKII), indicative of hypoxia, in perinecrotic zone. B and C. Lower concentrations of ATP (B) and glucose (C) in perinecrotic zone (p<0.05). D and E. No significant difference in glycogen (D) and lactate concentrations (E) between luminal and perinecrotic zone. n = 6, paired t-test.</p

Depletion of ATP, glucose and glycogen in advanced human plaques.

<p>A. Energy metabolites were analyzed in intermediate (CCA) and advanced (ICA) segments of human endarterectomies. B. Metabolite concentrations were assessed in the viable part of the intima (delineated), i.e. intimal area minus necrotic core. C, D and E. ATP (C), glucose (D) and glycogen concentrations (E) were lower in advanced segments than in intermediate segments of the same plaque. Note logarithmic scale for ATP and glycogen. F. Lactate concentrations were higher in advanced segments of the plaque. n = 6, Wilcoxon Signed-Rank Test.</p