Different Contributions of Physical Activity on Arterial Stiffness between Diabetics and Non-Diabetics

<div><p>Background</p><p>We compared the contribution of physical activity to the change in arterial stiffness between patients with and without diabetes in ischemic heart disease.</p><p>Methods</p><p>We studied 96 (diabetes) and 109 (without diabetes) patients with ischemic heart disease treated by percutaneous coronary intervention (PCI). Arterial stiffness was assessed by cardio-ankle vascular index (CAVI) at the first diagnosis of significant coronary ischemia and 6 months after PCI and optimal medical therapy. Physical activity was evaluated using the long form of the International Physical Activity Questionnaire (IPAQ).</p><p>Results</p><p>CAVI values increased more for diabetic patients than for non-diabetic. The IPAQ scores did not differ between the two groups. During follow-up, CAVI values did not significantly change in either group. In diabetic patients, the CAVI score for 48 patients did not change (NC-group) and 48 patients improved (Improved-group). Physical activity scores were 937.9 ± 923.2 and 1524.6 ± 1166.2 in the NC- and Improved-groups, respectively. IPAQ scores and uric acid levels significantly affect CAVI improvement after adjusting for age, sex, baseline CAVI, total cholesterol, and estimated glomerular filtration rate.</p><p>Conclusion</p><p>Determining factors influencing CAVI improvement during follow-up were significantly different between patients with and without diabetes. IPAQ scores and uric acid levels were significantly correlated with CAVI changes.</p></div

Odds ratio and 95% confidence interval of the improvement of CAVI during follow-up in patients with and without diabetes.

<p>Model adjusted for sex, age, blood pressure, the value of CAVI in the baseline, total cholesterol, eGFR, and uric acid.</p

Odds ratio (p-value) of the improvement of CAVI during follow-up.

<p>Odds ratio (p-value) of the improvement of CAVI during follow-up.</p

The ω-3 Polyunsaturated Fatty Acid, Eicosapentaenoic Acid, Attenuates Abdominal Aortic Aneurysm Development via Suppression of Tissue Remodeling

<div><p>Abdominal aortic aneurysm (AAA) is a prevalent vascular disease that can progressively enlarge and rupture with a high rate of mortality. Inflammation and active remodeling of the aortic wall have been suggested to be critical in its pathogenesis. Meanwhile, ω-3 polyunsaturated fatty acids such as eicosapentaenoic acid (EPA) are known to reduce cardiovascular events, but its role in AAA management remains unclear. Here, we show that EPA can attenuate murine CaCl₂-induced AAA formation. Aortas from BALB/c mice fed an EPA-diet appeared less inflamed, were significantly smaller in diameter compared to those from control-diet-fed mice, and had relative preservation of aortic elastic lamina. Interestingly, CT imaging also revealed markedly reduced calcification of the aortas after EPA treatment. Mechanistically, MMP2, MMP9, and TNFSF11 levels in the aortas were reduced after EPA treatment. Consistent with this finding, RAW264.7 macrophages treated with EPA showed attenuated <i>Mmp9</i> levels after TNF-α simulation. These results demonstrate a novel role of EPA in attenuating AAA formation via the suppression of critical remodeling pathways in the pathogenesis of AAAs, and raise the possibility of using EPA for AAA prevention in the clinical setting.</p></div

EPA reduces aortic aneurysm formation.

<p>Gross morphological and histological analyses of aortas were performed at 6 weeks after perivascular application of CaCl₂ to the infra-renal aorta. <b>A</b>. Representative images of <i>in situ</i> infra-renal aortas (demarcated by the broken lines) from mice in the sham-operated, control diet or EPA diet groups. <b>B</b>. Quantitative analysis of the maximal external aortic diameters of aortas. <i>n</i> = 4 for sham, <i>n</i> = 12 for control diet and EPA diet groups. <b>C</b>. Histological analysis by EVG staining, showing preserved aortic wall structure of the aorta from EPA diet group compared to the aorta from control diet group. Elastin breaks were also quantified. Scale bars, 200 µm (upper panels) and 50 µm (lower panels). <i>n</i> = 5 for sham, <i>n</i> = 11 for control diet, and <i>n</i> = 12 for EPA diet groups. Representative images of at least three independent experiments are shown in <b>A</b> and <b>C</b>. *<i>P</i><0.05.</p

EPA suppressed aortic calcification after AAA-induction.

<p>Aortic calcification was assessed by micro-CT imaging of <i>in situ</i> aortas 6 weeks after perivascular CaCl₂ application. Both sagittal and transverse slices (<b>A</b>) show reduced overall calcification in the infra-renal aortas from EPA diet group compared to the control diet group, and this was consistent with the results of quantitative analysis of the total calcification volume in each aorta (<b>B</b>). <i>n</i> = 4 for control diet and EPA diet groups. Red arrowheads indicate the posterior wall of the infra-renal aorta. Representative images of two independent experiments are shown in <b>A</b>. *<i>P</i><0.05 compared to control diet group.</p

MMP2, MMP9, and RANKL expression in AAAs.

<p><b>A</b>. Immunohistochemical staining for indicated proteins of serial sections of aortas one week after CaCl₂ treatment. Elastic van Gieson staining is also shown. SM α-actin and F4/80 were stained to locate SMCs and macrophages, respectively. Shown are representative images of 4 or more samples in each group. Scale bars, 50 µm. <b>B</b>. Relative positive staining area of MMP2, MMP9, and RANKL in sections from control diet and EPA diet groups. <i>n</i> = 4–5. *<i>P</i><0.05.</p

EPA attenuates <i>Mmp9</i> and <i>Tnfsf11</i> upregulation in CaCl₂-induced AAA. A

<p>. mRNA levels of the matrix metalloproteases <i>Mmp2</i> and <i>Mmp9</i>, as well as their tissue inhibitors <i>Timp1</i> and <i>Timp2</i>, in aortas at 1 and 3 weeks after perivascular CaCl₂ application were analyzed using real-time RT-PCR. <b>B</b>. The mRNA levels of the factors known to be involved in the development of vascular calcification, <i>Tnfsf11</i> and <i>Tnfrsf11b</i>, were also similarly analyzed using real-time RT-PCR. All expression levels were first normalized to <i>18s</i> rRNA levels and then presented as fold change over the sham group. *<i>P</i><0.05.</p

EPA reduces <i>Mmp9</i> expression in macrophages.

<p><b>A</b>. Gelatin zymography of aortic tissues one week after CaCl₂ treatment together with quantitative analysis, showing reduced MMP9 activity in samples from the EPA diet group. Equal amounts of protein (20 µg) were loaded per aortic sample. For quantitation, <i>n</i> = 6–7 in each group. <b>B</b>. Gating strategy for the flow cytometric analysis of AAA macrophages. Macrophages were identified as Ly-6C^lowCD11b⁺F4/80⁺Ly-6G⁻ cells (full gating strategy shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096286#pone.0096286.s001" target="_blank">Figure S2 in File S1</a>). <b>C</b>. The number of aortic macrophages per aortic sample. No statistically significant difference in the number of aortic macrophages between control diet and EPA diet groups was detected. <b>D</b>. The mRNA levels of <i>Mmp9</i> in sorted aortic macrophages. Expression levels were first normalized to <i>18s</i> rRNA levels and then further normalized to the level of control diet group. <i>n</i> = 5 in each group. <b>E</b>. RAW264.7 macrophages were cultured with either vehicle (10% BSA) or EPA (50 µmol/L) for 48 hours. The cells were then stimulated with recombinant mouse TNF-α (20 ng/mL) for a further 6 hours and harvested for analysis by RT-PCR. Expression levels were first normalized to <i>18s</i> rRNA levels and then presented as relative expression compared to baseline vehicle sample. <i>n</i> = 3 per condition. *<i>P</i><0.05 compared to control diet group in <b>A</b> and <b>D</b> or respective vehicle controls in <b>E</b>.</p

Echocardiography on day 21.

<p>FS, fractional shortening; EF, ejection fraction; IVSTd, interventricular septal thickness at end-diastole; LVIDd, left ventricular internal dimension diastolic; LVPWd, left ventricular posterior wall diastolic; LVIDs, left ventricular internal dimension systolic.</p><p>Echocardiography on day 21.</p