Atherosclerosis - Articles in Press

Cytokines release inhibition from activated monocytes, and reduction of in-stent neointimal growth in humans - Corrected Proof

2010-03-08

Abstract: Objective: Atherosclerosis and restenosis are largely ruled by inflammation. The aim of this study was to test the effects of a short-course, high-dose oral prednisone on the release of interleukin-6 (IL-6) and tumour necrosis factor (TNF)-α from circulating monocytes and on the neointimal growth that follows bare metal stent (BMS) implantation. In a sub-group of patients activated NF-κB was also evaluated.Methods: Out of 40 patients with coronary artery disease treated with BMS implantation, 20 were randomly assigned to receive oral prednisone during 40 days according to a standardized protocol. In non-stimulated and stimulated (LPS and PMA) monocytes we evaluated the release of IL-6 and TNF-α, and NF-κB p50 subunit translocation at baseline, at 10 and 30 days. Late luminal loss (LLL) 9 months after angioplasty was calculated by quantitative coronary angiography.Results: Plasma concentrations of prednisone correlated inversely with IL-6 and TNF-α release (R2=0.45, p=0.04 and R2=0.69, p=0.005, respectively) and NF-κB activation from monocytes (R2=0.58, p=0.01). The reduction of TNF-α release and NF-κB activation were significantly related (R2=0.56, p=0.01). Prednisone patients showed a significantly larger reduction of cytokine release and NF-κB activation compared to non-treated patients, at 10 days and 30 days. LLL was lower in the prednisone group (0.44±0.35mm versus 0.80±0.53mm, p=0.02) and correlated with reduction of TNF-α (R2=0.41, p=0.01).Conclusions: High doses of oral prednisone reduce NF-κB pathway activation and pro-inflammatory cytokine release in circulating activated monocytes of patients treated with coronary stenting. TNF-α release reduction correlates with decreased LLL.

Improvement of endothelial damage and regeneration indexes in patients with coronary artery disease after 4 weeks of statin therapy - Corrected Proof

2010-03-08

Abstract: Background: In patients with coronary artery disease (CAD), higher numbers of circulating endothelial progenitor cells (EPC) favourably influence clinical outcome. Controversially, increased apoptosis of endothelial cells (EC) may reflect vascular damage. Statins have been shown to improve vascular damage and enhance EPC function and numbers. The availability of ezetimibe, a potent novel cholesterol absorption inhibitor, allows to distinguish between lipid-lowering and pleiotropic properties of statins.Methods and findings: 43 patients with CAD were assigned to receive either: de novo atorvastatin (group A; n=17), ezetimibe as add-on to chronic statin therapy (group B; n=14), or dose escalation of atorvastatin (groupC; n=12) over 4 weeks. Circulating apoptotic EC (CD45−CD146+vWF+Annexin-V+) and EPC (CD34+KDR+) were quantified using flow cytometry.LDL cholesterol levels were significantly reduced in all treatment arms. Both statin groups, group A and group C, showed significantly reduced circulating apoptotic EC by 50% each (p<0.01). On the other hand, there was a significant doubling in the numbers of circulating EPC in group A and group C (p<0.005, each). Consequently, the endothelial damage-index calculated from numbers of circulating apoptotic mature EC related to EPC numbers, was improved in group A by 79% (p<0.01) and in group C by 70% (p<0.05). In contrast, sole LDL reduction by ezetimibe exerted no effect on any of the different circulating endothelial cell types.Conclusion: Thus, the improvement in numbers of EPC and reduction of mature apoptotic EC after 4 weeks of statin therapy, document a novel pleiotropic effect of statin therapy in patients with CAD.

Impact of HDL on adipose tissue metabolism and adiponectin expression - Corrected Proof

2010-03-04

Abstract: Objective: The objective of the current study was to investigate the hypothesis that high-density lipoprotein (HDL) influences adipocyte metabolism and adiponectin expression. Therefore, HDL was increased in vivo via apolipoprotein (apo) A-I gene transfer and in vitro via supplementation of HDL to partly differentiated adipocytes, in the presence or absence of lipopolysaccharide (LPS), known to decrease HDL cholesterol and adiponectin levels in vivo.Methods and results: Apo A-I transfer resulted in a significant increase of HDL cholesterol in control and LPS-injected C57BL/6 mice, which was paralleled by an increase in plasma adiponectin levels and adiponectin expression in abdominal fat. Triglyceride and free fatty acids levels after LPS administration were 2.2-fold (p<0.05) and 1.3-fold (p<0.05) lower, respectively, in Ad.hapoA-I-LPS than in Ad.Null-LPS mice. In parallel, the LPS-induced mRNA expression of hormone sensitive lipase was 3.5-fold (p=0.05) decreased in the Ad.hapoA-I-LPS group. On the other hand, apo A-I transfer abrogated the LPS-mediated reduction in lipin-1 and CD36 mRNA expression by 8.2-fold (p<0.05) and 18-fold (p<0.05), respectively. Concomitantly, the phosphorylation state of Akt was 2.0-fold (p<0.05) increased in the Ad.hapoA-I-LPS compared to the Ad.Null-LPS group. Pre-incubation of partly differentiated adipocytes with HDL (50μg protein/ml) increased adiponectin expression by 1.5-fold under basal conditions (p<0.05) and could abrogate LPS-induced down-regulation of adiponectin, both in a phosphatidylinositol-3-kinase-dependent manner.Conclusions: HDL affects adipocyte metabolism and adiponectin expression.

FDG–PET imaging of atherosclerosis: Do we know what we see? - Corrected Proof

Yuri Sheikine, Kamran Akram — 2010-03-04

Abstract: Imaging atherosclerosis may help to identify subjects harboring rupture-prone atherosclerotic plaques who may benefit from preventive interventions. Potential of plaques to rupture depends on their structural changes and metabolic activation, which are difficult to assess using anatomic imaging modalities. Recent studies suggested that functional imaging with positron emission tomography (PET) utilizing fluorine-18-labeled 2-deoxy-d-glucose (FDG) has the potential to assess plaque metabolism and add to prediction of vascular risk. Aortic, iliac, and carotid plaques can be detected with FDG–PET, even though not all plaques exhibit high FDG uptake. Detection of coronary artery plaques is more cumbersome due to technical limitations of PET and fast movement of these vessels during cardiac and respiratory cycles. Studies on substrate accumulating FDG in plaques are contradictory and mostly do not extend beyond correlation analyses. Vascular FDG uptake has an excellent short-term stability, but larger fluctuations of uptake long-term, which may complicate interpretation of such changes in therapeutic trials. FDG uptake in major arteries correlates with some cardiovascular risk factors and atherosclerosis markers, but clinical utility of such correlations is unclear. What is more important is that recently reported studies in cancer patients showed correlation between higher baseline FDG uptake and subsequent cardiovascular mortality. Anti-atherogenic therapy and therapeutic lifestyle changes seem to decrease vascular FDG uptake but it is not clear whether the latter predicts subsequent lower morbidity and mortality. These initial findings suggest that vascular FDG–PET may in the future find some utility in management of patients with atherosclerosis, but a number of important issues need to be addressed first. We need to: (1) determine optimal standard ways of performing imaging and quantifying vascular FDG uptake; (2) understand molecular mechanisms governing FDG accumulation in plaques; (3) perform studies prospectively linking vascular FDG uptake to cardiovascular events in non-cancer patients. As of today, vascular FDG–PET is not ready for its prime time in clinical practice.

What is the natural history of 18F-FDG uptake in arterial atheroma on PET/CT? Implications for imaging the vulnerable plaque - Corrected Proof

2010-03-04

Abstract: Purpose: Increased uptake of 18F-fluorodeoxyglucose (18F-FDG) in atherosclerotic plaque on Positron Emission Tomography (PET), predicts vulnerability. Recent studies have shown that the PET signal is reproducible over a 2-week period and as a result drug trials are underway. However, the natural history of these lesions is unknown. The aim of this study is determine the natural history of increased vascular wall uptake of 18F-fluorodeoxyglucose (18F-FDG).Methods: Following institutional ethics committee approval, we retrospectively examined PET/CT images of patients from our Institution that had at least 4 examinations in the last 5 years. This represented 205 studies in total, from 50 patients (29 men, 21 women, mean age 49.4±12.1 years, mean 5.1±1.7 studies/patient). The mean follow-up was 27.2±11.8 months. The carotids and the aorta were evaluated for increased 18F-FDG uptake with a maximum Standardized Uptake Value (SUVmax) >2.5, and >3.0, and calcification. Plots of SUVmax and Hounsfield units (HU) were made versus time.Results: The initial prevalence of increased focal arterial 18F-FDG uptake was 17/50 patients and of arterial calcification 19/50. 132 sites of 18F-FDG uptake in total were observed longitudinally. 18F-FDG vascular uptake did not persist with time. There was no correlation between 18F-FDG uptake and HU. No calcifications developed at sites of focal increased 18F-FDG uptake.Conclusions: Arterial lesions with increased 18F-FDG uptake represent transient phenomena. This data is important for the interpretation of findings of clinical trials using arterial 18F-FDG uptake as an imaging biomarker to monitor pharmacological intervention.

Mycophenolate mofetil attenuates plaque inflammation in patients with symptomatic carotid artery stenosis - Corrected Proof

2010-03-04

Abstract: Atherosclerosis as well as the subsequent progression towards cardiovascular events are considered to, at least partially, be a consequence of chronic inflammatory activity. Therefore, we decided to evaluate the impact of short-term immunosuppressive treatment on plaque characteristics in patients with symptomatic carotid artery stenosis.Twenty-one patients were randomized to receive either 1000mg. Mycophenolate mofetil (MMF) BD or placebo for at least 2 weeks prior to undergoing carotid endarterectomy (CEA). The serial sections of the CEA specimens were immunostained for activated T-cells (CD3+CD69+), regulatory T-cells (CD3+FOXP3+) and macrophages (CD68). In addition, gene expression profiling was performed by Illumina gene-array.Immunostaining revealed a reduction of activated T-cells in nine MMF-treated patients compared to 11 placebo-treated control patients (19.7% vs. 28.1%; p<0.05) as well as an increase of regulatory T-cells (3.8% vs. 1.8%; p=0.05). Microarray analyses confirmed beneficial changes to plaque phenotype, showing reduced expression of pro-inflammatory genes. Significantly reduced expression of metalloproteinases and osteopontin was observed in three out of nine MMF-treated patients compared to nil out of 11 in the placebo group.In the present study we show that immunosuppressive treatment for two-and-a-half weeks prior to CEA elicits changes in the plaque phenotype of symptomatic patients. These changes include reduced inflammatory cell presence with a concomitant decrease in pro-inflammatory gene expression.

Abnormal hepatic apolipoprotein B metabolism in type 2 diabetes - Corrected Proof

Bruno Vergès — 2010-03-02

Abstract: Increased Very Low Density Lipoprotein (VLDL) production is a major feature of diabetic dyslipidemia with consequences on the metabolism of other lipoproteins such as Low Density Lipoproteins (LDL) and High Density Lipoproteins (HDL). More precisely, we observe, in patients with type 2 diabetes, an increased production of VLDL1 particles that is potentially detrimental by generating atherogenic remnants, small dense LDL particles and triglyceride-rich HDL particles. Several pathophysiological factors are responsible for increased VLDL production, in type 2 diabetes. Among those, insulin resistance plays an important role. Indeed, defective activation of PI3-kinase, secondary to insulin resistance, is associated with a reduction of apoB degradation in the hepatocytes, a rise in MTP expression (by increasing nuclear transcription factors Fox01 and Foxa2) and an increased activity of phospholipase D1 and ARF-1, which are involved in VLDL1 formation. Moreover, peripheral insulin resistance is responsible for increased lipolysis of adipose tissue leading to augmented portal flux of FFA to the liver and, as a consequence, activation of VLDL production. In addition, increased de novo lipogenesis is observed in type 2 diabetes. This is secondary to increased activation of SREBP-1c (Sterol Regulatory Element-Binding Protein-1c), mainly by Endoplasmic Reticulum stress, and of ChREBP (Carbohydrate Responsive Element Binding Protein), mainly by hyperglycemia. Furthermore, decreased plasma adiponectin observed in type 2 diabetes, may also play a role in increased VLDL production by decreasing liver AMP-kinase activation and by increasing plasma FFA levels as a consequence of reduced muscle FFA oxidation.

Serum adipocyte fatty acid-binding protein is independently associated with coronary atherosclerotic burden measured by intravascular ultrasound - Corrected Proof

2010-03-02

Abstract: Objectives: Adipocyte fatty acid-binding protein (A-FABP) has been shown to have an effect on insulin resistance, lipid metabolism, and atherosclerosis in animals. We therefore investigated the association between the serum A-FABP level and coronary atherosclerosis.Methods: One hundred twenty-five consecutive patients with coronary artery disease (CAD) were enrolled after coronary angiography. Plaque volume in non-culprit coronary arteries was determined using intravascular ultrasound and expressed as percent plaque volume (%PV). Voluntary blood donors (n=120), matched for age and gender, served as controls. Serum levels of A-FABP, adiponectin, and inflammatory markers were measured by enzyme-linked immunosorbent assay.Results: The serum A-FABP level in CAD patients was significantly higher than in control subjects (median [25th–75th percentiles], 27.2 [20.5–37.1]ng/mL vs. 18.9 [14.6–24.5]ng/mL) (p<0.01). Serum A-FABP showed 0.74 of the area under the curve in the receiver operating characteristic curve for the detection of CAD, with 76% specificity and 65% sensitivity with a cut-off value of 20.1ng/mL. Further, in CAD patients, serum A-FABP had a significant correlation with %PV in all subjects (r=0.33, p<0.01). Serum A-FABP was positively correlated with the body mass index, serum interleukin-6 and high-sensitive CRP, and negatively correlated with HDL-cholesterol and serum adiponectin in CAD patients. Stepwise regression analysis revealed that serum A-FABP was independently associated with %PV.Conclusion: Increased serum A-FABP was significantly associated with a greater coronary plaque burden. Our findings revealed that the measurement of serum A-FABP could be utilized for the evaluation of the extent of coronary atherosclerosis.

Impact of plaque color by angioscopic evaluation on long-term clinical outcomes in patients with acute myocardial infarction undergoing percutaneous coronary intervention - Corrected Proof

Kazushi Saratani, Yoshitaka Iwanaga, Takahiro Hayashi, Shunichi Miyazaki — 2010-03-02

Abstract: Objective: This study assessed the relationship of in vivo plaque color in the culprit lesion by coronary angioscopy (CAS) with the long-term clinical outcome in patients after percutaneous coronary intervention (PCI) for acute myocardial infarction (AMI).Methods: Consecutive 224 patients with AMI were enrolled in the present study. The culprit lesions immediately before PCI were examined by CAS and were classified into three groups according to the color grade of the plaques: thick yellow plaque (TYP), yellow plaque (YP), or light yellow plaque (LYP).Results: There were no differences in baseline lipid and glycemic profiles and other coronary risk factors among the three groups. During the follow-up at a median of 4.9 years, the incidence of major adverse cardiac event (MACE) was significantly less in patients with TYP than those with YP or LYP by Kaplan–Meier analysis (p=0.027). TYP was an independent predictor for long-term MACE after adjustment of predictive factors (age, multivessel coronary artery disease, ejection fraction and body mass index, etc.).Conclusion: These results suggest that the TYP in culprit lesions was independently associated with the long-term favorable prognosis after AMI in patients treated with PCI. This relationship might provide new insights for pathophysiology of AMI and prevention of the secondary cardiac events after AMI.

Persistency of depression is associated with subclinical coronary atherosclerosis in males: Commentary on the study of Hamer et al. - Corrected Proof

Otavio C. Gebara, Raul D. Santos — 2010-03-02

The importance of psychosocial factors such as stress, anxiety, anger and depression, as risk factors for atherosclerotic coronary artery disease, has been investigated over the years . In the particular case of depression, which is one of the most prevalent psychiatric disorders, it has been shown that the presence of depressive symptoms was associated with worse cardiovascular prognosis in several studies, in different populations, with or without previous cardiovascular disease .

Leptin is associated with the size of the apolipoprotein(a) particle in African tribal populations living on fish or vegetarian diet - Corrected Proof

2010-03-02

Abstract: Objective: Apolipoprotein(a) [or apo(a)] isoform size, which is strongly genetically determined, showed significant association with the cardiovascular risk. Subjects on a fish diet have lower lipoprotein(a) levels, larger apo(a) isoform sizes and lower leptin levels than their vegetarian diet counterparts. We hypothesized that leptin may contribute to a potential association between the type of diet and the size of apo(a) isoforms.Methods: Anthropometric data, dietary nutrients, lipoprotein profile, plasma leptin levels, and apo(a) isoforms were evaluated in two related homogenous African tribal populations of Tanzania, one on a primarily freshwater fish diet (n=278), and the other on a vegetarian diet (n=326).Results: We observed a strong negative association between leptin levels and size of each of the apo(a) isoforms in both fish and vegetable diet groups, and in both genders. However, leptin was not associated with levels of lipoprotein(a). In multivariate analysis, a strong and independent association between leptin and size of apo(a) isoforms was observed. The size of apo(a) isoforms was strongly associated with high and low leptin states. Subjects with low leptins had 30% larger sizes of apo(a) isoforms than their high leptin counterparts.Conclusions: High leptin subjects have smaller, potentially more atherogenic, apo(a) isoform sizes than low leptin ones. We suggest that omega-3 rich diet can influence the levels of apo(a) and/or Lp(a) even though they are mainly genetically determined. These findings may have implications for understanding the interaction between leptin and cardiovascular risk.

Systemic MCP1/CCR2 blockade and leukocyte specific MCP1/CCR2 inhibition affect aortic aneurysm formation differently - Corrected Proof

2010-03-02

Abstract: Objective: CCR2, the receptor for monocyte chemoattractant protein 1 (MCP1), is involved in atherosclerosis and abdominal aortic aneurysms (AAAs). Here, we explored the potential beneficial blockade of the MCP1/CCR2 pathway.Methods: We applied an AAA model in aging apolipoprotein E deficient mice with pre-existing atherosclerotic lesions. These mice were subjected to two therapeutic strategies. First, a dominant negative form of MCP1 was overexpressed in femoral muscles, resulting in circulating levels of MCP1-7ND (7ND), competing with native MCP1. In the second approach, bone marrow transplantation was performed using bone marrow cells that were infected with a lentiviral construct containing siRNA for CCR2, to specifically inhibit only leukocyte CCR2 expression.Results: Both strategies did not influence lesion size of the advanced atherosclerotic plaques. However, 7ND induced a more fibrous plaque phenotype. Yet, surprisingly a trend in increased number and severity of AAA was observed in the 7ND group. Smooth muscle cells in the aneurysm showed decreased phosphorylated signal transducer and activator of transcription five (STAT5, P<0.01) in the 7ND group, which is indicative for a decreased proliferative and migratory (wound healing) response. This presumably resulted in the increased AAA development. In contrast, siRNA-induced inhibition of CCR2 in leukocytes led to a significant inhibition in aneurysm formation. In conclusion, systemic inhibition of the MCP1/CCR2 pathway leads to a fibrous plaque phenotype in the advanced atherosclerotic lesions, but to potential adverse effects on AAA formation, implying that for a beneficial overall therapeutic approach, specific inhibitory targeting of leukocyte CCR2 will be essential.

Evaluation of the lipid lowering ability, anti-inflammatory effects and clinical safety of intensive therapy with Zhibitai, a Chinese traditional medicine - Corrected Proof

2010-03-02

Abstract: Objectives: To evaluate the lipid lowering ability, anti-inflammatory effects and clinical safety of intensive therapy of the Chinese traditional medicine Zhibitai in subjects with moderate to high cardiovascular risk.Methods: A total of 169 subjects (96 males and 73 females, aged 55–72) having moderate to high cardiovascular risk were recruited and randomly divided into Zhibitai group (n=85), which received 480mg of Zhibitai orally twice daily, and atorvastatin group (n=84), which received 10mg of atorvastatin orally once a day. Blood lipoproteins, myocardial enzymes, liver and renal functions were measured before treatment started, and after 4 and 8 weeks of the treatment. High sensitivity C-reactive protein (hs-CRP), P-selectin, matrix metalloproteinase-9 (MMP-9) and soluble intercellular adhesion molecule-1 (sICAM-1) were measured before and after the treatment.Results: Plasma total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) were significantly decreased, and high-density lipoprotein cholesterol (HDL-C) was increased in both groups, after 4 and 8 weeks of treatment (p<0.05 for all pairs). Interestingly, plasma triglycerides (TG) decreased in the Zhibitai group after 4 weeks of treatment but only decreased in the atorvastatin group after 8 weeks. Inflammatory factors such as hs-CRP, P-selectin, MMP-9 and sICAM-1 were significantly decreased in both groups after 8 weeks (p<0.01 for all pairs). Furthermore, there was no difference in myocardial enzymes, hepatic and renal function test parameters, incidence of myopathy or gastrointestinal tract symptoms in either group.Conclusion: Zhibitai therapy is a good alternative to statin therapy to reduce plasma cholesterol levels in subjects with moderate to high cardiovascular risk. Most importantly, Zhibitai is safe to use.

Paraoxonase 1 deficiency in mice is associated with reduced steroid biosynthesis: Effects on HDL binding, cholesteryl ester accumulation and scavenger receptor type BI expression - Corrected Proof

2010-03-02

Abstract: Objective: Selective uptake of high density lipoprotein (HDL) cholesteryl ester (CE) is considered as the major source of cholesterol for production of steroids in the adrenal gland in rodents. As paraoxonase 1 (PON1) is an HDL-associated lipo-lactonase that has been shown to increase binding of HDL to macrophages, we used PON1 knock-out (PON1KO) mice to test the possible role of PON1 in corticosterone (CS) biosynthesis.Methods and results: PON1 deficiency was associated with reduced serum CS concentration. Adrenal glands obtained from PON1KO mice had significantly lower CE content compared to adrenals from C57Bl6 control mice. Binding of HDL obtained from PON1KO mice to human adrenocortical carcinoma cell line was found to be significantly lower than that of control HDL, and was associated with decreased CS biosynthesis. Addition of purified PON1 to HDL from PON1KO mice increased HDL binding and CS synthesis. Furthermore, the expression of the HDL receptor, SR-BI, protein and mRNA, was reduced in adrenals from PON1KO mice compared to control mice. When challenged with low salt diet, PON1KO mice demonstrated an increase in adrenal SR-BI gene expression and in serum corticosterone which reached levels similar to those obtained in control mice.Conclusion: PON1 regulates adrenal CS biosynthesis at two levels: (a) via an accessory role in HDL binding properties, and (b) a supportive role in SR-BI expression and CE supply to the cells.

Characterization of culprit lesions in acute coronary syndromes using coronary dual-source CT angiography - Corrected Proof

2010-03-02

Abstract: Objective: We analyzed typical morphological features of coronary atherosclerotic plaques in acute coronary syndromes (ACS) using contrast-enhanced coronary Dual-Source CT angiography (CTA) in comparison to stable coronary lesions.Patients and methods: Fifty-five patients with ACS and 55 controls with stable angina pectoris (SAP) with similar atherosclerotic risk profile were studied. CT angiography was performed using a Dual-Source CT scanner (330ms rotation, 2×64×0.6mm collimation, 60–80mL contrast agent i.v. at 6mL/s) before invasive catheterization. We analyzed plaque volume (mm3), mean and minimal CT density (HU), remodeling index, plaque type (calcified/non-calcified/mixed) and presence of “spotty” calcifications as well as presence of contrast rims.Results: In patients with ACS and SAP, 28 and 10 lesions showed both calcified and non-calcified components, but in a greater proportion of non-calcified material, 6 and 23 lesions showed a greater proportion of calcified material and 21 and 8 lesions were completely non-calcified, respectively. None of the culprit plaques in ACS and 14 of the lesions in SAP were completely calcified. A “spotty” pattern of calcification within the plaque and a central filling defect surrounded by a rim of contrast were present in 11 and 14 of 55 ACS cases, but never in SAP lesions. For culprit lesions in ACS and for lesions in patients with SAP, mean plaque volumes were 192.8±114.9mm3 and 103.8±51.8mm3 (p=0.001), mean and minimal CT densities were 85.6±45.1HU and 47.2±33.7HU versus 143.8±104.1HU and 95.9±84.0HU (p<0.01) and mean remodeling indices were 1.6±0.4 and 0.97±0.17 (p<0.001), respectively.Conclusion: Plaques of culprit lesions in ACS show specific morphologic characteristics in non-invasive coronary CT angiography. As compared to stable lesions, culprit lesions in ACS display greater proportion of non-calcified material and presence of “spotty” calcifications/contrast rims as well as larger plaque volumes, lower CT attenuation and higher remodeling indices.

Accumulation and expression of serum amyloid P component in human atherosclerotic lesions - Corrected Proof

Zhiqing Song, Lei Cai, Ling Guo, Yoshitane Tsukamoto, Chikao Yutani, Xiang-An Li — 2010-03-02

Abstract: Serum amyloid P component (SAP) is a member of pentraxins. Previous studies indicate that SAP exists in human atherosclerotic aortic intima and the plasma SAP levels are associated with cardiovascular disease. In this study, we characterized SAP in normal and atherosclerotic intima, investigated the source of SAP in atherosclerotic lesions, and assessed the effect of SAP on HDL function. Immunohistochemical staining and electroimmunoassay indicated that SAP is not present in normal aortic intima which excludes the possibility that SAP non-specifically deposits in aortic intima via its binding to microfibrils. Notably, SAP levels are correlated with the severity of atherosclerotic lesions. Fast protein liquid chromatography (FPLC) and Western blot analysis revealed that SAP exists in atherosclerotic lesions in multiple forms. Soluble SAP accumulates in the lesions as decamer in free or bound forms via ligand-binding to its ligand(s). Insoluble SAP accumulates in the lesions in covalent-bound forms conjugated to collagen/collagen-like substances via disulfide (–S–S–) bonds. In situ hybridization and RT-PCR analysis revealed that SAP is generated in atherosclerotic lesions, at least partly, by macrophages and smooth muscle cells in neointima. Functional analysis demonstrated that SAP associated with HDL promotes SR-BI-dependent cholesterol efflux and lipid-free SAP enhances ABCA1-dependent cholesterol efflux. In conclusion, our findings demonstrate that SAP is specifically accumulated and expressed in atherosclerotic lesions. SAP may be involved in cholesterol clearance through its role in promoting cholesterol efflux.

Can the extent of epicardial adipose tissue thickness or the presence of descending thoracic aortic calcification predict significant coronary artery stenosis in patients with a zero coronary calcium score on multi-detector CT? - Corrected Proof

2010-03-02

Abstract: Objective: The purpose of this study was to evaluate whether the extent of epicardial adipose tissue (EAT) thickness or the presence of descending thoracic aortic calcification on multi-detector CT (MDCT) can predict the presence of significant coronary artery stenosis in patients with negative coronary artery calcium (CAC).Methods: We enrolled 90 patients with negative CAC in whom both coronary CT angiography (CTA) and conventional angiography had been performed. Group 1 consisted of patients (n=27) with significant coronary artery stenosis (≥50%), whereas group 2 (n=63) had non-obstructive coronary artery stenosis (<50%) on conventional angiography. We analyzed whether or not there is a significant difference in EAT thickness or the incidence of calcification of descending thoracic aorta among the two groups.Results: There was no significant difference between EAT thickness on MDCT among the two groups. There was also no significant difference in the incidence of descending thoracic aortic calcification between group 1 (7/27, 25.9%) and group 2 (14/63, 22.2%) (p>0.05).Conclusions: Neither the presence of abundant EAT nor calcification of descending thoracic aorta is a marker of significant coronary artery stenosis in patients with negative CAC.

Impact of age, sex and exercise on brachial and popliteal artery remodelling in humans - Corrected Proof

2010-03-02

Abstract: Objective: To examine the impact of age, sex and exercise on wall thickness and remodelling in the popliteal and brachial arteries.Methods: We compared wall thickness, lumen diameter and wall:lumen ratios in the brachial and popliteal arteries of 15 young (Y, 25.4±0.8yr; 7M 8W) and 16 older sedentary (OS, 58.8±1.1yr; 8M 8W) subjects, with 12 of the OS group also studied following 12 and 24 weeks exercise training.Results: Wall thickness and lumen diameter were higher in the popliteal than the brachial artery for both groups (P<0.05); wall:lumen ratio was similar between arteries. Comparison of the Y and OS groups revealed no impact on wall thickness, whereas diameter values were higher in OS subjects (P<0.05). Whilst there were no significant differences in wall thickness between men and women in the Y or OS groups, diameter was larger in men than in women for both arteries (P<0.05). After 24 weeks of training the wall thickness of both arteries decreased (P<0.01) and the wall:lumen ratio of the brachial (P<0.01) and the popliteal (P<0.05) decreased.Conclusion: The cross-sectional results suggest that ageing was associated with increased lumen diameter, although wall:lumen ratio remained unchanged. Wall:lumen ratio was higher in women than men, irrespective of subject age or the artery studied. This related primarily to differences in lumen diameter between the sexes, as wall thickness did not significantly differ between men and women. Our longitudinal data strongly suggest that exercise training is associated with beneficial effects on conduit artery wall thickness and wall:lumen ratio in both upper and lower limbs in humans.

Lysophospholipids: Effectors mediating the contribution of dyslipidemia to calcification associated with atherosclerosis - Corrected Proof

Germán Camejo — 2010-03-02

Calcification nodules in the extracellular intima of atherosclerotic plaques are a common finding in human arteries and in vessels of animal models in which lesions are induced by lipoprotein abnormalities. In addition, medial diffuse calcification is also associated with atherosclerosis co-existing with chronic kidney dysfunction or diabetes. Minor diffuse calcification in aging arteries may be of no clinical relevance. However, intimal and medial prominent calcium deposits that can protrude into the arterial lumen induce abnormal hemodynamic patterns and changes of the vessel wall visco-elastic properties. These calcification-associated flow disturbances can contribute to plaque progress through responses of the subendothelial extracellular matrix (ECM) . Calcifications in arteries can also be accelerated by dyslipidemias and already in 1976 Hollander proposed a unified concept for the link of calcium accumulation and lipoprotein entrapment on the polysaccharides of the ECM . Intimal calcification takes place mostly within collagen fibers that are embedded in a tridimensional mesh of proteoglycans (PG). This is a milieu created mainly by smooth muscle cells and also the site of apoB-lipoproteins association with PG, a phenomenon that appears to be one of the initial steps in atherogenesis . Therefore, elucidation of the molecular events linking intimal calcification with ECM properties and lipoprotein retention is an important research area in atherosclerosis. In the current issue of Atherosclerosis Vickers, Castro-Chavez and Morriset (reference) have shown that lysolecithin, probably generated from LDL deposited in the intima, can be one of the external signals triggering the phenotypic change of arterial smooth muscle cells (ASMC) leading to calcification in a lipid rich environment.

Cigarette smoking and beneficial effect on cardiometabolic risk: A reply - Corrected Proof

Altan Onat — 2010-02-25

We were the first to show 3 years ago prospectively that cigarette smoking protected Turkish adults from metabolic syndrome (MetS) and type-2 diabetes . The mechanism appeared to be largely mediated by reducing obesity and/or by influence on inflammatory mediators. Since then we described that protection from incident hypertension , association with lower serum complement C3 and apo C-III concentrations and in women, particularly diminished or no effect on future C-reactive protein levels , reduced accumulation of fat in the visceral region , reduced serum pre-heparin LPL mass combined with a certain genotype and other mechanisms were also involved in the favorable effect of smoking on cardiometabolic risk among Turks. This population is characterized by a high prevalence of MetS, diabetes and coronary heart disease. All these cardiometabolic disorders are associated with enhanced pro-inflammatory state and/or oxidative stress. In such an environment, the effect of smoking on cardiometabolic risk in Turkish men is less hazardous than in Westerners and, as shown by our findings among women, may even be beneficial. It has been shown that, i.e. the process of nicotine acting as a prominent angiogenic agent , via using endothelial acetylcholinergic receptors, is enhanced in milieus of inflammation and ischemia .

The effects of silencing EDF-1 in human endothelial cells - Corrected Proof

Marzia Leidi, Massimo Mariotti, Jeanette A.M. Maier — 2010-02-25

Abstract: Objective: EDF-1, a 16kDa highly conserved intracellular protein, serves as a calmodulin binding protein and, upon nuclear translocation, functions as a coactivator of several transcription factors. To understand whether EDF-1 is implicated in regulating endothelial function, we silenced EDF-1 expression using small hairpin (sh) RNA.Methods: Human umbilical vein endothelial cells (HUVEC) were utilized and EDF-1 levels were detected by western blot. Cell proliferation, cell organization in fibrin gel and nitric oxide release were evaluated in cells silencing EDF-1 after transfection with shRNA.Results: EDF-1 was downregulated in quiescent and senescent HUVEC, whereas it was upregulated in proliferating cells. Knocking down EDF-1 promoted the acquisition of a spindle phenotype, inhibited cell proliferation, accelerated the organization into capillary-like networks on fibrin gels and induced the production of nitric oxide (NO). While the total amounts and the degree of phosphorylation of endothelial NO synthase are not altered in cells silencing EDF-1, we found an increased interaction between calmodulin and endothelial NO synthase. Accordingly, the calmodulin inhibitor calmidazolium significantly decreased NO release in cells silencing EDF-1. These results suggest that knocking down EDF-1 might increase free calmodulin which ultimately activates endothelial NO synthase.Conclusions: Since EDF-1 (i) is involved in the control of endothelial proliferation and organization, events which are crucial to repair damages to the vessel wall, and (ii) increases NO, which exerts anti-atherogenic and anti-thrombotic effects, we conclude that EDF-1 is implicated in molecular events that are pivotal to endothelial function and, therefore, to vascular integrity.

5′-Nitro-indirubinoxime inhibits inflammatory response in TNF-α stimulated human umbilical vein endothelial cells - Corrected Proof

2010-02-25

Abstract: Inflammation plays a critical role in the development of atherosclerosis and TNF-α, a major inflammatory cytokine, induces inflammatory responses by enhancing the expression of adhesion molecules and the secretion of inflammatory mediators. Indirubin is an active compound of Polygonum tinctorium Lour (P. tinctorium) that has the ability to suppress inflammation. Previously, we described the novel indirubin derivative, 5′-nitro-indirubinoxime (5′-NIO), and demonstrated that it has potent anti-proliferative activity against various human cancer cells. In this study, we examined the effect of 5′-NIO on the TNF-α induced inflammatory conditions of human umbilical vein endothelial cells (HUVECs). We found that 5′-NIO inhibited TNF-α induced MCP-1 and IL-8 expression at the RNA and protein levels in HUVECs. Specifically, 5′-NIO significantly inhibited the TNF-α stimulated release of MCP-1 and IL-8, with levels that were only 19.8% and 30.9% of those of untreated control cells, respectively. Furthermore, 5′-NIO largely inhibited the adhesion of U937 cells to HUVECs by decreasing the expression level of ICAM-1 and VCAM-1. Overall, these observations suggest that 5′-NIO has the potential for use as an anti-atherosclerotic agent.

Coronary flow reserve is impaired in patients with nonalcoholic fatty liver disease: Association with liver fibrosis - Corrected Proof

2010-02-24

Abstract: Background: Nonalcoholic fatty liver disease (NAFLD) is associated with an increased risk of cardiovascular disease. Coronary flow reserve (CFR) is widely used to examine the integrity of coronary microvascular circulation. We evaluated the prevalence of impaired CFR in patients with biopsy-proven NAFLD. We also investigated the independent clinical, biochemical, and liver histology predictors of CFR in the setting of NAFLD.Methods: Fifty-nine consecutive patients with NAFLD and 77 age- and gender-matched controls were evaluated. CFR recordings were performed by transthoracic Doppler harmonic echocardiography. CFR≥2.0 was considered normal.Results: CFR was significantly lower in patients with NAFLD than in controls (2.11±0.45 vs. 2.52±0.62, P<0.001). An impaired CFR (i.e. <2) was found in 25 NAFLD patients (42.4%) whereas all controls had normal CFR values (P<0.001). A stepwise linear regression analysis in NAFLD patients identified liver fibrosis scores as the only independent predictor of CFR values (β=−0.60; t=−2.44, P=0.021).Conclusion: Our findings indicate that in patients with biopsy-proven NAFLD: (a) an abnormal CFR is found in approximately 42.4% of cases, and (b) liver fibrosis scores are an independent predictor of depressed CFR.

Genetic variations at ABCG5/G8 genes modulate plasma lipids concentrations in patients with familial hypercholesterolemia - Corrected Proof

2010-02-22

Abstract: Objective: To investigate the association of four common single nucleotide polymorphisms (SNPs) at ABCG5 (i7892A>G, i18429C>T, Gln604GluC>G, i11836G>A) and five at ABCG8 (5U145T>G, Tyr54CysA>G, Asp19HisG>C, i14222T>C, and Thr400LysG>T) with plasma lipids concentrations and to explore the interaction between those SNPs and smoking in patients with FH.Methods and Results: ABCG5/G8 SNPs were genotyped in 500 subjects with genetic diagnosis of FH. Carriers of the minor A allele at the ABCG5_i11836G>A SNP displayed significantly higher HDL-C concentrations (P=0.023) than G/G subjects. In addition, carriers of the minor G allele at the ABCG5_Gln604GluC>G SNP had significantly lower VLDL-C (P=0.011) and lower TG (P=0.017) concentrations than homozygous C/C. Interestingly, a significant gene-smoking interaction was found, in which carriers of the minor alleles at ABCG5 (i7892A>G, i18429C>T, i11836G>A) SNPs displayed significantly lower HDL-C, higher TC and higher TG respectively, only in smokers. On the other hand, nonsmokers carriers of the minor alleles at ABCG5 (i18429C>T and Gln604GluC>G) SNPs had significantly lower TG concentrations (P=0.012 and P=0.035) compared with homozygous for the major allele.Conclusions: Our data support the notion that ABCG5/G8 genetic variants modulate plasma lipids concentrations in patients with FH and confirm that this effect could be influenced by smoking. Therefore, these results suggest that gene-environmental interactions can affect the clinical phenotype of FH.

Preclinical carotid atherosclerosis enhances the global cardiovascular risk and increases the rate of cerebro- and cardiovascular events in a five-year follow-up - Corrected Proof

2010-02-22

Abstract: Aim: To evaluate if the intima-media thickening (IMT) and asymptomatic carotid plaque (ACP), as expression of carotid preclinical atherosclerosis (pre-ATS), can provide further information on the global cardiovascular risk (GCVR).Methods: We studied 454 asymptomatic subjects, with a cluster of risk factors (RF), and evaluated the incidence of a first cardiovascular (CV) event in a five-year follow-up. The subjects at admission were subdivided in three groups of risk.Results: Events occurred in 38% of subjects at high risk, in 13% and 6% of subjects at intermediate and low risk (p<0.003). Among evaluated parameters, carotid pre-ATS was a predictive marker of CV events (OR 2.7, 95% IC 1.4–5.1, p<0.0024). In subjects with GCVR <20% the prevalence of events was 8% for normal carotid ultrasound findings, 13% for increased IMT and 15% for ACP.Conclusions: In primary prevention, the IMT measurement can give further information for a better stratification of GCVR. The pre-ATS of carotid arteries should be considered a strong predictor of future CV events and should suggest a more aggressive treatment of RF.

Association of selected ABC gene family single nucleotide polymorphisms with postprandial lipoproteins: Results from the population-based Hortega study - Corrected Proof

2010-02-22

Abstract: The aim of the study was to determine the influence of twenty single nucleotide polymorphisms (SNPs) of the ABCA1, ABCG1, ABCG5 and ABCG8 genes on the plasmatic concentrations of total cholesterol (TC), HDL and LDL cholesterol (HDLc, LDLc) in the postprandial state with a representative Spanish Caucasian population (1473 individuals, 50.0% women, ages ranging 21–85 years). In men, subjects with the AA genotype of the ABCA1 rs2230806 (R219K) polymorphism were associated with increased plasma LDLc levels, while the ABCA1 haplotype, which included the rs2230806 A allele, was associated with higher TC and LDLc plasma concentrations. In women, significant relationships were found between rs1893590 polymorphisms (ABCG1 gene) and HDLc plasma concentrations (subjects with the AA genotype had lower HDLc levels). For the ABCG8 gene, the rs4148211 polymorphism was associated with higher plasma TC and LDLc concentrations in the total population. Moreover, an ABCG5-G8 haplotype, which included the rs6544718 T allele, was associated with higher HDLc plasma concentrations in women. In conclusion, different SNPs of the ABCA1, ABCG1 and ABCG5-ABCG8 genes were associated, some under gender-specific analysis, with variations in the plasma lipid levels under postprandial conditions in a representative Spanish population.

Sulfatides are associated with neointimal thickening after vascular injury - Corrected Proof

2010-02-22

Abstract: Background: Sulfatides are known to be a native ligand of P-selectin. Platelet-leukocyte interaction via the cross-talk between P-selectin and Mac-1 (CD11b/CD18) plays an important role in the mechanism of neointimal thickening after vascular injury such as that seen in post-stent restenosis. However, the roles of sulfatides on restenosis have not been elucidated.Methods: Serum sulfatide levels, P-selectin expression on the surface of platelets, and activated Mac-1 on the surface of neutrophils were serially measured using both coronary sinus and peripheral blood samples in 21 patients who underwent coronary stent implantation.Results: The trans-cardiac gradient (coronary sinus minus peripheral blood) of the sulfatide levels significantly increased at 15min (−1.47±2.87 to 0.59±1.44nmol/ml, p<0.001), compared to baseline levels. The maximum response of the trans-cardiac gradient of P-selectin expression on the surface of platelets at 15min after stent implantation (R=0.55, p<0.01), and that of activated Mac-1 on the surface of neutrophils at 48h (R=0.59, p<0.01), were both positively correlated with that of serum sulfatide levels at 15min. The angiographic late lumen loss was correlated with the trans-cardiac gradient of sulfatide levels at 15min (R=0.48, p<0.05), platelet P-selectin expression at 15min (R=0.42, p<0.05) and activated neutrophil Mac-1 expression at 48h (R=0.46, p<0.05), but not with values at other sampling points.Conclusions: Sulfatides may play a physiological role on inflammation in vascular injury and the development of neointimal thickening.

Protective effects of magnesium lithospermate B against diabetic atherosclerosis via Nrf2-ARE-NQO1 transcriptional pathway - Corrected Proof

2010-02-22

Abstract: Hyperglycemia-induced oxidative stress is known to play an important role in the development of several diabetic complications, including atherosclerosis. Although a number of antioxidants are available, none have been found to be suitable for regulating the oxidative stress response and enhancing antioxidative defense mechanisms. In this study, we evaluated the effects of magnesium lithospermate B (LAB) against oxidative stress. We also endeavored to identify the target molecule of LAB in vascular smooth muscle cells (VSMCs) and the underlying biochemical pathways related to diabetic atherosclerosis. Modified MTT and transwell assays showed that the increased proliferation and migration of rat aortic VSMCs in culture with high glucose was significantly inhibited by LAB. LAB also attenuated neointimal hyperplasia after balloon catheter injury in diabetic rat carotid arteries. To determine molecular targets of LAB, we studied the effects of LAB on aldose reductase (AR) activity, O-GlcNAcylation, and protein kinase C (PKC) activity in VSMCs under normoglycemic or hyperglycemic conditions and showed the improvement of major biochemical pathways by LAB. Potential involvement of the nuclear factor erythroid 2-related factor-2 (Nrf2) – antioxidant responsive element (ARE)-NAD(P)H: quinone oxidoreductase-1 (NQO1) pathway was assessed using siRNA methods. We found that LAB activates the NQO1 via the Nrf2-ARE pathway, which plays an important role in inhibition of the major molecular mechanisms that lead to vascular damage and the proliferation and migration of VSMCs. Together, these findings demonstrate that the induction of the Nrf2-ARE-NQO1 pathway by LAB could be a new therapeutic strategy to prevent diabetic atherosclerosis.

Decreased number and impaired functionality of endothelial progenitor cells in subjects with metabolic syndrome: Implications for increased cardiovascular risk - Corrected Proof

I. Jialal, S. Devaraj, U. Singh, B.A. Huet — 2010-02-22

Abstract: Objective: Metabolic syndrome (MetS) is characterized by low-grade inflammation and confers an increased risk for cardiovascular disease. Endothelial progenitor cells (EPCs) are a measure of vascular health and are decreased in patients with various risk factors for cardiovascular disease (CVD). There is a paucity of data examining the EPC status especially in terms of their functionality in MetS subjects without diabetes or cardiovascular disease. We aimed to enumerate and functionally characterize EPCs in subjects with MetS in comparison to healthy controls.Methods: The study was performed at the University of California Davis Medical Center. Healthy controls (n=31) and MetS (n=46) subjects were included in the study. EPCs were enumerated in fasting blood by KDR/CD34 dual positivity. Functionality was assessed by the colony forming units (CFU) assay, migration and tubule formation.Results: Subjects with MetS had significantly decreased number of EPCs compared to control subjects. Furthermore, EPCs from MetS subjects depicted significantly impaired clonogenic capacity, i.e., decreased colony forming units, and impaired capacity to incorporate into tubular structures suggesting functional impairment of EPCs from MetS subjects.Conclusions: We make the novel observation that MetS subjects without diabetes or CVD have decreased EPC number and impaired functionality as compared to control subjects. These findings could contribute to the increased CV risk in this population.

Epidemiology of TAO – Correction of an error - Corrected Proof

Raphael Adar — 2010-02-19

Quoting from unchecked references may be hazardous to your reputation. The review by Malecki et al. on thromboangiitis obliterans (TAO) is a superb up-to-date treatise covering all aspects of this enigmatic disease . However, I feel compelled to point out a gross error in the section on Epidemiology. Commenting on the incidence of TAO among patients with peripheral vascular disease (PVD) in different ethnic groups, the authors quote an 80% incidence among Ashkenazi Jews.

High-resolution ultrasound showing increased intima and media thickness of the radial artery in patients with end-stage renal disease - Corrected Proof

Mats Johansson, Anna Myredal, Peter Friberg, Li Ming Gan — 2010-02-18

Abstract: Objective: Although clinically relevant atherosclerosis of the upper limb arteries is rarely seen, intimal hyperplasia of the arteries may reflect global atherosclerosis and increased intima-media thickness of the brachial artery has been linked to an increased risk of cardiovascular events and to early failure of the radiocephalic arteriovenous fistula. We speculated that patients with ESRD have thickening of both the radial intimal and medial layers compared to healthy subjects.Methods: Ultrasound biomicroscopy is a novel very high frequency (55MHz) ultrasound technique that could accurately measure the intima and media thickness of the vessel wall. No previous study has measured intima and media thickness separately in patients with end-stage renal disease and hence, the aim of the current study was to investigate the radial arterial wall layers in patients with chronic renal failure.Results: Thirty-one patients with end-stage renal disease and 41 healthy subjects underwent ultrasound biomicroscopy of the radial arteries. Blood pressures did not differ except for pulse pressures which were elevated among patients with end-stage renal disease (p<0.01). Patients with end-stage renal disease showed 39% thicker intima and 18% greater media in the radial artery compared to healthy subjects (0.117±0.031mm versus 0.084±0.02mm for the IT, p<0.01 and 0.205±0.062mm versus 0.174±0.044mm for the MT; p<0.05).Conclusions: Both the intima and the media layers of the radial arteries are increased in patients with end-stage renal disease. Whether measurements of the radial arterial intima thickness may convey valuable information on the risk of future cardiovascular events and early arteriovenous fistula failure in end-stage renal disease remain to be elucidated in future studies.

Variability of residual platelet function despite clopidogrel treatment in patients with peripheral arterial occlusive disease - Corrected Proof

2010-02-15

Abstract: Residual platelet function despite treatment with clopidogrel may predict an unfavourable cardiovascular outcome. The majority of studies have investigated the effects of clopidogrel administration in conjunction with aspirin in patients undergoing percutaneous coronary intervention. The primary objective of the present study was to assess the platelet response to clopidogrel in the absence of aspirin in patients with peripheral arterial occlusive disease (PAOD) and to investigate whether non-responsiveness to clopidogrel is reproducible during long-term follow-up. Fifty-four clinically stable PAOD patients on a maintenance dose of 75mg/d clopidogrel were enrolled in this study. Platelet function was assessed at baseline and after a median follow-up of 18 months using light transmittance aggregometry (LTA) with 2μM ADP as an agonist. HPLC-coupled mass spectrometry was used to detect clopidogrel and clopidogrel carboxylic acid, the main metabolite of clopidogrel. Residual platelet function, as defined by late aggregation values within the reference range (i.e., >43%), was observed in 35.2% of patients at baseline and 17.6% during follow-up. During the observation period, 26.5% had switched from responder to non-responder status or vice versa. Among non-responders, either clopidogrel or its metabolite was detected in 89.5% and 83.3% of patients at baseline and at follow-up, respectively. We conclude that non-responsiveness to clopidogrel as determined by ADP-induced LTA is not stable over time. This phenomenon cannot be attributed to non-compliance alone.

Association of plaque in the carotid and coronary arteries, using MDCT angiography - Corrected Proof

2010-02-15

Abstract: Objective: Associations between various plaque types in coronary and carotid arteries using multidetector row computed tomography (MDCT) have not yet been reported. We evaluated MDCT correlation of total plaque and various plaque subtypes in carotid and coronary arteries.Methods: We studied 62 patients who had both carotid and cardiac computed tomography angiography (CTA). The plaque in each vascular segment was classified as non-calcified, calcified or mixed.Results: The average age of this population was 68±22 years, 63% males. Total plaque in the carotid artery correlated with total, calcified and mixed plaque in the coronary artery (all P<0.001). Calcified plaque in the carotid artery was associated with total and calcified plaque in the coronary bed (P<0.001). Non-calcified coronary plaque was not associated with carotid plaque.Conclusion: Total plaque in coronary and carotid arteries are highly correlated.

ESR1 genetic variants, haplotypes and the risk of coronary heart disease and ischemic stroke in the Finnish population: A prospective follow-up study - Corrected Proof

2010-02-15

Abstract: Association of estrogen receptor 1 (ESR1) gene variants and risk of coronary heart disease (CHD) and ischemic stroke was evaluated in the FINRISK-study. From 14,140 individuals, 2225 were selected for genotyping using a case-cohort design. Time-to-event analysis showed that the CC genotype of −397T/C ERS1 gene contributed to higher risk of CHD only in men (HR, 1.68, CI 1.03–2.74). The −351A/G polymorphism was not independently associated with CHD. Haplotype analysis of these two variants indicated that in men, haplotype TA conferred lower risk of CHD (HR=0.72, CI 0.55–0.95), whereas men with haplotype CA had 1.8 higher risk of CHD events (CI 1.21–2.77), compared to other haplotypes. No association was found with ischemic stroke. Our study suggests that the minor allele −397C of the ESR1 gene confers risk of CHD among Finnish men, both in homozygous state and as part of a haplotype with the −351A allele.

Persistent cognitive depressive symptoms are associated with coronary artery calcification - Corrected Proof

Mark Hamer, Mika Kivimaki, Avijit Lahiri, Michael G. Marmot, Andrew Steptoe — 2010-02-15

Abstract: Objectives: The association between depression and sub-clinical atherosclerosis remains unclear. By assessing depressive symptoms only at one point in time, most previous studies have failed to ascertain long-term exposure. We examined the association of long-term depressive symptoms assessed at three time points (over 10 yrs) with a marker of sub-clinical atherosclerosis.Methods: Participants included 454 healthy, non-medicated men and women from the Whitehall II epidemiological cohort without known cardiovascular disease (CVD). Depressive symptoms were assessed at three time points (over 10 yrs) and coronary atherosclerosis was assessed at follow-up in terms of coronary artery calcification (CAC).Results: 18.9% of the sample reported depressive symptoms at least once during follow-up. Participants that were persistently depressed had over a two-fold increased risk of detectable CAC (Agatston score>0) (odds ratio [OR]=2.56, 95% CI, 1.14–5.78) and high CAC (Agatston score≥100) (OR=2.36, 1.04–5.35) compared with never depressed after adjustment for age, sex, and a range of conventional cardiac risk factors. These associations were more robust in men. Participants who were depressed on only one occasion were not at elevated risk of CAC.Conclusions: Persistent cognitive symptoms of depression assessed over several time points, but not on a single occasion, are related to sub-clinical coronary atherosclerosis in men free of known CVD and diabetes.

Peri-aortic fat, cardiovascular disease risk factors, and aortic calcification: The Framingham Heart Study - Corrected Proof

2010-02-12

Abstract: Objective: Perivascular fat through the secretion of paracrine and pro-inflammatory mediators may play a role in obesity-mediated vascular disease. We sought to examine associations between adipose tissue depots immediately surrounding the thoracic aorta, metabolic risk factors, and vascular calcification.Methods: In participants free of cardiovascular disease (CVD) from the Framingham Heart Study Offspring cohort who underwent computed tomography (n=1067, mean age 59 years, 56.1% women), thoracic peri-aortic fat depots were quantified. Visceral abdominal tissue (VAT) and calcification of the thoracic and abdominal aorta were also measured.Results: Peri-aortic fat depots were correlated with body mass index, waist circumference (WC), VAT (all p<0.0001), hypertension (p=0.007), low HDL (p<0.0001), serum triglycerides (p<0.0001), impaired fasting glucose (p=0.005), and diabetes (p=0.02). These associations generally remained significant after adjustment for BMI and WC (all p-values<0.05), but not after VAT adjustment. Thoracic aortic fat was associated with thoracic calcification in models containing VAT (OR 1.31, 95% CI 1.01–1.71, p=0.04), but was not significant after adjustment for CVD risk factors (OR 1.16, 95% CI 0.88–1.51, p=0.30). Thoracic aortic fat, however, was associated with abdominal aortic calcification (OR 1.48, 95% CI 1.11–1.98, p=0.008) and coronary artery calcification (OR 1.47, 95% CI 1.09–1.98, p=0.001) even in models including CVD risk factors and VAT.Conclusions: Thoracic peri-aortic fat is associated with measures of adiposity, metabolic risk factors, and coronary and abdominal aortic calcification.

Elevated heart rate and cardiovascular outcomes in patients with coronary artery disease: Clinical evidence and pathophysiological mechanisms - Corrected Proof

2010-02-12

Abstract: There is an established body of evidence from epidemiological studies which indicates that an elevated resting heart rate is independently associated with atherosclerosis and increased cardiovascular morbidity and mortality, in both the general population and in patients with established cardiovascular disease. Clinical trial data suggest that in patients with coronary artery disease, an elevated heart rate identifies those at increased risk of adverse cardiovascular outcomes, and that lowering of heart rate may reduce major cardiovascular events in patients with an elevated heart rate and symptom-limiting angina. These results suggest that an increased heart rate may have an adverse impact on the atherosclerotic process and increase the risk of a cardiovascular event in patients with coronary artery disease. The precise pathophysiological mechanisms that link heart rate and cardiovascular outcomes have yet to be defined. Possibilities may include indirect mechanisms related to autonomic dysregulation and those due to an increase in heart rate per se, which can increase the ischaemic burden and exert local haemodynamic forces that can adversely impact on the endothelium and arterial wall. For these reasons, heart rate should be considered as a therapeutic target in the treatment of patients with coronary artery disease.

Peak radial and circumferential strain measured by velocity vector imaging is a novel index for detecting vulnerable plaques in a rabbit model of atherosclerosis - Corrected Proof

2010-02-10

Abstract: Aims: To evaluate the reliability of velocity vector imaging (VVI) for detecting vulnerable plaques.Methods and Results: After aortic balloon injury, 60 rabbits were fed a 1% cholesterol diet for 10 weeks and normal chow for another 6 weeks. Adenovirus containing p53 or lac Z was then injected into the aortic plaques and rabbits were divided into p53-treated group (n=20), lac Z-treated group (n=20) and blank control group (n=20). Peak longitudinal (LSp), radial (RSp) and circumferential (CSp) strain of plaques was measured using VVI at the end of week 18 before pharmacological triggering. Higher RSp and CSp and lower LSp were found in ruptured than those in non-ruptured plaques, and RSp, CSp and LSp correlated well with the fibrous cap thickness and plaque content of macrophages, smooth muscle cells and collagen (all p<0.01). A logistic regression model showed that both RSp (RR: 8.96, 95% CI: 5.3575–10.4857, p<0.001) and CSp (RR: 8.45, 95% CI: 5.9043–9.1043, p<0.001) were significant predictors of plaque rupture. RSp and CSp had a sensitivity of 88.0% and 88.6% and a specificity of 88.6% and 92.0% to predict plaque disruption, respectively.Conclusion: VVI offers a new and noninvasive technique for measuring the peak strain of atherosclerotic plaques and RSp and CSp are a novel index with a high sensitivity and specificity for detecting plaques vulnerable to rupture.

The renal artery ostium flow diverter: Structure and potential role in atherosclerosis - Corrected Proof

Edward B. Neufeld, Zu-Xi Yu, Danielle Springer, Qing Yu, Robert S. Balaban — 2010-02-10

Abstract: Initiation of renal atherosclerosis occurs primarily at the caudal region of the renal artery ostium. To date, no mechanism for initiation of atherosclerosis at this site has been substantiated. Herein, we identify a renal artery flow diverter on the caudal wall of the renal artery ostium that directs flow into the renal artery and selectively retains LDL, an initial step in atherosclerosis. High-resolution ultrasound revealed the generation of flow eddies by the caudal diverter in vivo, consistent with a role in directing aortic flow to the renal artery. Two-photon excitation en face microscopy of the diverter revealed a substantial reduction in the elastic lamina exposing potential retention sites for LDL. Fluorescent LDL was selectively retained by the renal artery diverter, consistent with its molecular structure. We propose that the rigid macromolecular structure of the renal artery ostium diverter is required for its vascular function and contributes to the initiation of renal atherosclerosis by the retention of LDL.

Paraoxonase 1 (PON1) deficiency in mice is associated with reduced expression of macrophage SR-BI and consequently the loss of HDL cytoprotection against apoptosis - Corrected Proof

Bianca Fuhrman, Anna Gantman, Michael Aviram — 2010-02-10

Abstract: Background: Paraoxonase 1 (PON1) was shown to stimulate HDL binding and HDL-mediated cholesterol efflux from macrophages. This study examined the role of PON1 in the expression of proteins that enhance macrophage HDL binding, i.e. ABCA1 and SR-BI.Methods and results: ABCA1 expression was similar, whereas SR-BI expression (mRNA and protein determined by FACS, Western blot, or immunocytochemistry) was significantly decreased in peritoneal macrophages from PON1 deficient (MPM-PON10) in comparison to C57Bl/6 (MPM-Control) mice. PON1 deficiency correction with HDL-control, recombinant PON1 (rePON1), or by transfection with a plasmid containing the rePON1 gene, increased SR-BI expression in MPM-PON10, whereas rePON1/H115Gln mutant, or the H115Q/H134Q double mutant, which lack catalytic activity, did not stimulate SR-BI expression. Lysophosphatidyl choline (LPC) resulting from PON1 action on macrophage PC, upregulated SR-BI expression in MPM-PON10 via activation of ERK1/2 and PI3K. Functionally, HDL bound to MPM-PON10 significantly less than to MPM-Control, and failed to inhibit tunicamycin-induced apoptosis, but had no significant effect on HDL-mediated cholesterol efflux from macrophages.Conclusions: PON1 deficiency in mice is associated with decreased macrophage SR-BI expression, decreased cellular HDL binding, and consequently the loss of HDL-mediated cytoprotection against apoptosis, which may contribute to the accelerated atherosclerosis observed in PON10 mice. These findings add new insights into the function of SR-BI in macrophages, and define the potential role of PON1 in regulating SR-BI-mediated HDL protection against macrophages apoptosis.

Relationship between Mediterranean Diet Score and atherothrombotic risk: Findings from the Third National Health and Nutrition Examination Survey (NHANES III), 1988–1994 - Corrected Proof

Stephanie J. Carter, Mary B. Roberts, Jason Salter, Charles B. Eaton — 2010-02-08

Abstract: Background: Mediterranean diet has been promoted as the preferred dietary model for cardiovascular disease prevention in the United States.Objective: We sought to evaluate the degree to which the Mediterranean diet is associated with reduced levels of atherothrombotic biomarkers in a population-based sample in the U.S.Design: Data from 13,197 adults between the ages of 18 and 90 were collected and atherothrombotic risk factors assessed as part of the NHANES III, 1988–1994. Adherence to the Mediterranean diet was evaluated using food frequency questionnaires, supplemented by the 24-h dietary recall data, to develop Mediterranean Diet Scores (MedDietScore) that were analyzed in tertiles. The cross-sectional relationship of MedDietScore to atherothrombotic factors were analyzed using multiple variable regression analysis adjusted for complex sampling design using SUDAAN.Results: The components of the Mediterranean diet and the dietary pattern's associations with atherothrombotic risk factors differed by age and gender. For men <45 years of age as MedDietScore increased: total cholesterol/HDL cholesterol (TC/HDL) ratio (p=0.0390), serum insulin (p=0.0414), and white blood cell (WBC) (p=0.0246) decreased. For men ≥45 years as MedDietScore increased: TC/HDL ratio (p=0.0008), Hemoglobin A1c (HgbA1c) (p=0.0001), HOMA index (p=0.0486), C-reactive protein (p=0.0034), fibrinogen (p=0.0028) decreased and HDL cholesterol (HDL-c) levels (p<0.0001) increased. For pre-menopausal women, as MedDietScore increased: TC/HDL ratio (p<0.0001), non-HDL cholesterol (p=0.0012), apolipoprotein B (p=0.0112), HgbA1c (p=0.0001), decreased and HDL-c levels (p<0.0001) increased. For post-menopausal women, as MedDietScore increased: TC/HDL ratio (p=0.0005), Triglycerides (p<0.0001), serum insulin (p=0.0062), HOMA index (p=0.0063) and Homocysteine (Hcy) (0.0046) levels decreased and HDL-c levels (p=0.0005) increased.Conclusions: Mediterranean diet appears to be associated with selective measures of cardioprotective lipid profiles, glucose metabolism, and inflammation and coagulation levels.

Are serum cholesterol levels associated with silent brain infarcts? The Seiryo Clinic Study - Corrected Proof

2010-02-08

Abstract: Objective: High levels of serum cholesterol are associated with the risk of stroke. However, the association of serum cholesterol with silent brain infarcts (SBIs) is unclear. We investigated the association between SBI and various clinical factors.Methods: We conducted a cross-sectional study that included 324 apparently healthy Japanese men (mean age 53.8±9.2 years). Combinations of three types of scan (T1-weighted, T2-weighted and FLAIR images) were used to detect and discriminate SBI.Results: Serum cholesterol was significantly associated with SBI [total cholesterol, odds ratio (OR) 3.75 (95% confidence interval (CI) 1.45–9.68); LDL-cholesterol, OR 2.54 (95% CI 1.03–6.27), and non-HDL-cholesterol, OR 2.54 (95% CI 1.03–6.27)] after adjustment for age, smoking status, serum triglycerides, maximal-intima-media thickness, obesity, hypertension, diabetes mellitus, hyperuricemia, coronary heart disease and lipid-lowering agent use.Conclusion: Our cross-sectional data suggest that serum cholesterol levels are associated with SBI independently of known confounders.

Cholesterol diet and effect of long-term withdrawal on plaque development and composition in the thoracic aorta of New Zealand White rabbits - Corrected Proof

2010-02-08

Abstract: Aims: Experimental study on plaque progression, regression and composition in atherosclerotic thoracic aorta of hypercholesterolemic rabbits after long-term withdrawal of cholesterol-enriched diet (CED).Methods: Rabbits were fed 2% cholesterol for 6 weeks followed by withdrawal periods for 15, 23, 34, 68, or 78 weeks. Cholesterol, triglyceride, and phospholipids levels in blood and cholesterol concentrations in aorta were quantified. Plaque size and cellularity, phenotype of macrophages and smooth muscle cells were (immuno)histomorphometrically analyzed in segments of the thoracic aorta.Results: After 6 weeks of CED, blood cholesterol levels were about 80-fold higher, whereas atherosclerosis and cholesterol content in the thoracic aorta were only minimally increased. However, the latter significantly increased within 15 weeks after cholesterol withdrawal, while serum cholesterol level was still 10-fold increased. Thereafter plaque area and cholesterol content remained almost unchanged until the end of the study despite a long-term normalization of serum cholesterol level after withdrawal of CED. Directly after 6 weeks of CED the densities of macrophages and apoptotic cells within plaques were highest, decreasing after cholesterol withdrawal, whereas, vice versa the density of smooth muscle cells (SMCs) significantly increased.Conclusion: We suggest that atherosclerotic plaques respond to long-term withdrawal of CED by decrease in number and phenotype of macrophages and increase of SMCs without regression of the lesion size. The cellular changes are suggested to considerably contribute to higher plaque stability.

ABC transporters, atherosclerosis and inflammation - Corrected Proof

Michael L. Fitzgerald, Zahedi Mujawar, Norimasa Tamehiro — 2010-02-08

Abstract: Atherosclerosis, driven by inflamed lipid-laden lesions, can occlude the coronary arteries and lead to myocardial infarction. This chronic disease is a major and expensive health burden. However, the body is able to mobilize and excrete cholesterol and other lipids, thus preventing atherosclerosis by a process termed reverse cholesterol transport (RCT). Insight into the mechanism of RCT has been gained by the study of two rare syndromes caused by the mutation of ABC transporter loci. In Tangier disease, loss of ABCA1 prevents cells from exporting cholesterol and phospholipid, thus resulting in the build-up of cholesterol in the peripheral tissues and a loss of circulating HDL. Consistent with HDL being an athero-protective particle, Tangier patients are more prone to develop atherosclerosis. Likewise, sitosterolemia is another inherited syndrome associated with premature atherosclerosis. Here mutations in either the ABCG5 or G8 loci, prevents hepatocytes and enterocytes from excreting cholesterol and plant sterols, including sitosterol, into the bile and intestinal lumen. Thus, ABCG5 and G8, which from a heterodimer, constitute a transporter that excretes cholesterol and dietary sterols back into the gut, while ABCA1 functions to export excess cell cholesterol and phospholipid during the biogenesis of HDL. Interestingly, a third protein, ABCG1, that has been shown to have anti-atherosclerotic activity in mice, may also act to transfer cholesterol to mature HDL particles. Here we review the relationship between the lipid transport activities of these proteins and their anti-atherosclerotic effect, particularly how they may reduce inflammatory signaling pathways. Of particular interest are recent reports that indicate both ABCA1 and ABCG1 modulate cell surface cholesterol levels and inhibit its partitioning into lipid rafts. Given lipid rafts may provide platforms for innate immune receptors to respond to inflammatory signals, it follows that loss of ABCA1 and ABCG1 by increasing raft content will increase signaling through these receptors, as has been experimentally demonstrated. Moreover, additional reports indicate ABCA1, and possibly SR-BI, another HDL receptor, may directly act as anti-inflammatory receptors independent of their lipid transport activities. Finally, we give an update on the progress and pitfalls of therapeutic approaches that seek to stimulate the flux of lipids through the RCT pathway.

The epidemiology of subclavian stenosis and its association with markers of subclinical atherosclerosis: The Multi-Ethnic Study of Atherosclerosis (MESA) - Corrected Proof

2010-02-08

Abstract: Background: Recent studies indicate that subclavian stenosis (SS), diagnosed by a large systolic blood pressure difference (SBPD) between the right and left brachial arteries, is associated with cardiovascular disease (CVD) risk factors and outcomes. We sought to describe the epidemiology of SS and determine its association with markers of subclinical CVD in the baseline cohort of the Multi-Ethnic Study of Atherosclerosis.Methods: We defined SS by an absolute SBPD ≥15mmHg. Peripheral artery disease (PAD) was defined by an ankle-brachial index ≤0.90. The coronary artery calcium score (CAC) and the common carotid artery intima-media thickness (CCA-IMT) were measured by computed tomography and B-mode ultrasound, respectively. Odds ratios for the associations of SS with risk factors and subclinical disease were estimated using logistic regression.Results: Of 6743 subjects studied, 307 participants (4.6%) had SS, with a higher prevalence in women (5.1%) than men (3.9%), and in African Americans (7.4%) and non-Hispanic whites (5.1%) than Hispanic (1.9%) or Chinese (1.0%) participants (p<0.01). In a model including age, gender, ethnicity, traditional and novel CVD risk factors, significant associations with SS were observed for C-reactive protein (highest vs. three lower quartiles: OR=1.41; 95%CI: 1.06–1.87) and brachial artery pulse pressure (OR=1.12/10mmHg; 95%CI: 1.03–1.21). Adjusted for age, gender, ethnicity, traditional and novel CVD risk factors, SS was significantly associated with PAD (OR=2.35; 1.55–3.56), with CCA-IMT (highest vs. the lower three quartiles: OR=1.32; 1.00–1.75), and high CAC (score >100 vs. score=0; OR=1.43; 1.03–2.01).Conclusions: The subclavian stenosis is positively associated with other markers of subclinical atherosclerosis.

Reduced LDL-cholesterol levels in patients with coronary artery disease are paralelled by improved endothelial function: An observational study in patients from 2003 and 2007 - Corrected Proof

2010-02-08

Abstract: Objective: Recent guidelines recommend more aggressive lipid-lowering in secondary prevention protocols. We examined whether this resulted in improved endothelial function.Methods: We studied saphenous vein specimens of patients undergoing surgical coronary revascularisation in 2007 and compared results with those of patients examined in 2003. Endothelium-dependent vasodilation was assessed by relaxation to calcium ionophore A23187, and vascular superoxide production by lucigenin enhanced chemiluminescence.Results: Statin dose increased from 26±16mg/d in 2003 to 37±17mg/d in 2007 (P<0.001), and total (4.0±0.9mmol/L vs 4.8±1.0mmol/L) and LDL-cholesterol levels (2.0±0.7mmol/L vs 3.0±0.9mmol/L) were lower in 2007 compared to 2003 (P<0.001; n=90 each). Endothelium-dependent vasodilation was greater in 2007 (44±15%) compared to 2003 (28±12%; n=36 each; P<0.001). Vascular superoxide derived from endothelial NO synthase (eNOS) was lower in 2007 than in 2003 (reduction by NG-nitro-l-arginine-methyl ester, 0.29±0.21nmol/(mgmin) vs 0.09±0.20nmol/(mgmin); P=0.002). In linear regression analysis, LDL-cholesterol levels have been shown to be the major determinant of endothelial function in the combined 2003 and 2007 cohort.Conclusion: Intensive lipid-lowering is associated with improved endothelial function and reduced superoxide production from eNOS. Further improvement in vascular function could be achieved by targeting other sources of superoxide including xanthine oxidase.

Tocotrienol enriched palm oil prevents atherosclerosis through modulating the activities of peroxisome proliferators-activated receptors - Corrected Proof

Fengjuan Li, Wenjuan Tan, Zhanfang Kang, Chi-Wai Wong — 2010-02-08

Abstract: Palm oil is enriched in vitamin E in the form of α-, γ-, and δ-tocotrienols. Dietary tocotrienol supplements have been shown to prevent atherosclerosis development in patients and preclinical animal models. However, the mechanistic basis for this health beneficial effect is not well established. Peroxisome proliferator-activated receptors α, γ, and δ (PPARα, PPARγ, and PPARδ) are ligand regulated transcription factors that play essential preventive roles in the development of atherosclerosis through regulating energy metabolism and inflammation. In this study, we presented data that the tocotrienol rich fraction (TRF) of palm oil activated PPARα, PPARγ, and PPARδ in reporter based assays. Importantly, TRF attenuated the development of atherosclerosis in ApoE−/− mice through inducing PPAR target gene liver X receptor alpha (LXRα) and its down-stream target genes apolipoproteins and cholesterol transporters, suggesting that modulating the activities of PPARs is a key aspect of the in vivo action of tocotrienols.

Estrogen attenuates vascular remodeling in Lp(a) transgenic mice - Corrected Proof

2010-02-08

Abstract: Objective: Although it is well known that Lipoprotein(a) (Lp(a)) is an atherogenic lipoprotein and an independent risk factor for cardiovascular disease, there is no confirmed therapy to decrease Lp(a) or prevent atherosclerosis induced by Lp(a). Thus, it is mandatory to develop novel therapy to prevent atherosclerosis in high Lp(a) concentration. Here, we focused on the effect of estrogen on Lp(a) level and Lp(a)-induced vascular remodeling.Methods: We employed Lp(a) transgenic mice (human apo(a) yeast artificial chromosome (YAC) and human apoB double transgenic mice). Vascular remodeling was induced by ligation of the common carotid artery and the effect of estrogen was evaluated in female mice after ovariectomy with or without estrogen replacement.Results: Estrogen deficiency caused by ovariectomy increased serum Lp(a), and continuous replacement of 17β-estradiol (20μg/kg/day) reversed the change. In the vascular remodeling model induced by carotid artery occlusion, neointima formation was significantly increased in ovariectomized female Lp(a) transgenic mice, but few in male Lp(a) transgenic mice, as compared to wild FVB mice. Importantly, continuous replacement of estrogen in ovariectomized mice significantly attenuated it. In cultured endothelial cells and macrophages, addition of Lp(a) increased mRNA of ICAM-1, VCAM-1, E-selectin and MCP-1 in endothelial cells and TNF-α, IL-1β and MCP-1 in macrophages in a dose-dependent manner. Importantly, pre-treatment with estrogen attenuated these changes in a dose-dependent manner.Conclusion: Estrogen negatively regulates both plasma Lp(a) level and Lp(a)-induced vascular remodeling, suggesting that estrogen might be a strong candidate to reduce serum Lp(a) concentration.

Decreased endothelin-1 levels after acute consumption of red wine and de-alcoholized red wine - Corrected Proof

2010-02-08

Abstract: Background: Red wine consumption may influence on vasoconstrictive peptide endothelin-1 levels, and this may be one mechanism leading to improved vasodilation after red wine consumption. Endothelin-1 levels and their association with coronary epicardial diameter and flow rate, however, have not been studied in vivo after consumption of red wine and de-alcoholized red wine. The purpose of this randomized trial was to determine the acute effects of these beverages on endothelin-1 levels and compare them to coronary artery epicardial diameter and flow rate.Methods: Twenty-two healthy men consumed a high dose (8.1±0.9dL) of alcohol-containing red wine and de-alcoholized red wine in a cross-over design at one sitting with a two-week washout period. Endothelin-1 levels were determined and coronary artery diameter and flow rate assessed using transthoracic echocardiography before and acutely after intervention.Results: Red wine and de-alcoholized red wine significantly decreased endothelin-1 levels (0.75±0.26pg/mL to 0.61±0.20pg/mL, p=0.002; 0.74±0.32pg/mL to 0.63±0.24pg/mL, p=0.04, respectively), but did not have a significant effect on epicardial diameter (1.1±0.3mm vs. 1.1±0.3mm, p=0.58; and 1.1±0.3mm vs. 1.1±0.2mm, p=0.10, respectively) or flow rate (7.8±4.0mL/min to 6.4±3.6mL/min, p=0.07; and 7.8±4.0mL/min to 7.4±3.2mL/min, p=0.53, respectively).Conclusions: Red wine and de-alcoholized red wine decreased plasma endothelin-1 levels after acute consumption, but this change was not reflected in coronary epicardial diameters or flow rate.

Mycophenolic acid attenuates tumor necrosis factor-α-induced endothelin-1 production in human aortic endothelial cells - Corrected Proof

2010-02-08

Abstract: Aims: Atherosclerotic cardiovascular disease is the major cause of morbidity and mortality in solid organ transplant recipients. Endothelin-1 (ET-1) is implicated in the pathogenesis of atherosclerosis and is one of the potential therapeutic targets. This study was conducted to evaluate the effect of mycophenolic acid (MPA), an immunosuppressant for the transplant recipients, on tumor necrosis factor-α (TNF-α)-induced ET-1 production in aortic endothelial cells.Methods and results: In cultured human aortic endothelial cells, TNF-α increased ET-1 through AP-1 and NF-κB, whereas MPA attenuated it by reducing both AP-1 and NF-κB DNA-binding activities. TNF-α increased ET-1 via c-Jun NH2-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK), but not extracellular signal-regulated kinase. N-acetylcysteine that downregulated TNF-α-induced reactive oxygen species (ROS) inhibited JNK activation, but not p38 MAPK. N-acetylcysteine, SP600125 (JNK inhibitor) and SB203580 (p38 MAPK inhibitor) attenuated TNF-α-induced DNA-binding activities of both AP-1 and NF-κB. MPA inhibited JNK and p38 MAPK activations as well as ROS generation. N-acetylcysteine, SP600125, SB203580 and MPA had no effect on either TNF-α-induced IκBα degradation or p65 nuclear translocation, but attenuated p65 Ser276 phosphorylation.Conclusion: MPA attenuated TNF-α-induced ET-1 production through inhibitions of ROS-dependent JNK and ROS-independent p38 MAPK that regulated NF-κB as well as AP-1. These findings suggest that MPA could have an effect of amelioration of atherosclerosis.