Abstract
Background and design
Methods
Results
Conclusion
Keywords
Introduction
In contrast to moderate drinking, heavy and binge drinking (five or more drinks for men and four or more for women per occasion) are known to increase mortality from several causes, including cardiovascular diseases [1]. The mechanisms of increased cardiovascular risk associated with heavy drinking remain unclear, nor is it known whether all beverages are equally harmful in high doses.
Apart from traditional atherosclerosis risk factors, other risk factors — such as oxidative stress, endothelial cell activation, inflammation, factors regulating fibrinolysis and thrombosis, and activation of plaque proteolytic enzymes — are now recognized as important contributors in the disease's progression and complications. Nevertheless, the levels of these soluble markers of atherosclerosis have not been previously assessed before and after drinking high doses of red wine, dealcoholized red wine and cognac.
The purpose of this randomized controlled cross-over study was to determine the acute effects of heavy drinking of red wine, dealcoholized red wine and cognac on circulatory markers of atherosclerosis.
Methods
The study included 22 healthy, nonsmoking, normolipidemic, and nondiabetic Finnish men (mean age 23 ± 1.8 years, body mass index 24 ± 2.3 kg/m2) who were not taking any medication [2]. The study was carried out in accordance with the Declaration of Helsinki (2000) of the World Medical Association and approved by the Ethics Committee of the Southwest Finland Health Care District. All participants gave their written informed consent.
The participants were randomly assigned in a cross-over design to take doses of red wine, dealcoholized red wine or cognac after overnight fasting on separate days with a washout period of at least 1 week. The doses of red wine and cognac contained 1.0 g/kg of ethanol. Dealcoholized red wine was ingested in volumes equal to those of red wine. The full doses of red wine, dealcoholized red wine, and cognac were 8.1 ± 0.9, 8.1 ± 0.9, and 2.4 ± 0.4 dl, respectively.
Perlage Cabernet del Veneto 2003 (Italy), Ebony Vale Cabernet Sauvignon 2003 (grapes from Italy; dealcoholization in Germany), and Remy Martin VSOP (France) were chosen as the red wine, dealcoholized red wine, and cognac, respectively. Phenolic profiles were determined using an analytical high-performance liquid chromatography method as described earlier [3, 4]. Flavonoids (anthosyans, cathecins, oligomeric procyanidines, flavonols) and other phenolic substances (resveratrol) were analyzed in their natural form. Serum ethanol concentrations were determined 1-2 h after the ingestion of the high dose of red wine and cognac using a standard photometric test (Modular P800, Roche Diagnostics GmbH, Mannheim, Germany). Concentrations of plasma soluble E-selectin, soluble vascular cell adhesion molecule 1, soluble intercellular adhesion molecule 1, serum matrix metalloproteinase 9, serum myeloperoxidase, serum adiponectin, tissue plasminogen activator inhibitor-1 (tPAI-1), and high-sensitivity C-reactive protein concentrations were measured before and 1-2 h after each beverage consumption in duplicate using LINCO-plex Human Cardiovascular Disease Panel 1 Kit 96 Well Plate Assay (Millipore Cat. HCVD1-67AK, Missouri, USA) and the Luminex 200 and Luminex XYP platform (Luminex, Austin, Texas, USA).
The data are presented as means ± SD. A paired t-test and Bonferroni's method (altogether 24 addressed hypotheses) were used to compare the serum levels before and after interventions and between beverages. P values of less than 0.05 were considered significant. Statistical analyses were done using SPSS 15.0 (SPSS Inc., Chicago, Illinois, USA).
Markers of vascular endothelial cell activation (sE-selectin, sVCAM-1, sICAM-1), Inflammation (hsCRP), oxidative stress (MPO), fibrinolysis (tPAI-1), plaque destabilization (MMP-9), and adlponectln before and after the Ingestion of the beverages
CI, confidence interval; hsCRP, high-sensitivity C-reactive protein; MMP-9, matrix metalloproteinase 9; MPO, myeloperoxidase; sE-selectin, soluble E-selectin; sICAM-1, soluble intercellular adhesion molecule 1; sVCAM-1, soluble vascular cell adhesion molecule 1; tPAI-1, tissue plasminogen activator inhibitor-1.
Results
Red wine, unlike dealcoholized red wine and cognac, increased tPAI-1 levels significantly (Table 1). Moreover, a significant difference was found in the change of tPAI-1 levels between red wine and dealcoholized red wine (P < 0.001), and red wine and cognac (P < 0.01), but not between dealcoholized red wine and cognac. A decreasing trend in matrix metalloproteinase 9 levels after cognac ingestion was observed, but not after red wine or dealcoholized red wine ingestion. The markers high-sensitivity C-reactive protein, E-selectin, soluble vascular cell adhesion molecule 1, soluble intercellular adhesion molecule 1, myeloperoxidase, and adiponectin remained unchanged after the interventions.
Serum alcohol levels increased to 1.3 ± 0.2 and 1.2 ± 0.2% after the ingestion of high doses of red wine and cognac, respectively. Resveratrol was abundant only in red wine. Phenolic substances — anthosyans, flavonols, and cathecins — were detected in red wine and dealcoholized red wine but not in cognac.
Discussion
This randomized controlled cross-over study shows that drinking of high doses of red wine increased tPAI-1 levels, indicating enhanced inhibition of fibrinolysis, unlike similar doses of dealcoholized red wine and cognac. The increase after red wine ingestion is not explained only by the ethanol content of red wine, because the amount of ethanol was similar when cognac was ingested.
This was the first time that soluble markers related to atherosclerosis were tested in a prospective randomized clinical study after ingestion of red wine, its dealcoholized form and cognac. It seems that some of the phenolic substances present in red wine may cause excessive increase of tPAI-1 levels. Our results suggest that these may include resveratrol, cathecins, or anthosyans, which are all present in red wine, but not in reasonable concentrations in cognac or dealcoholized red wine. A previous observation that drinking more than four glasses of red wine a day induced an inhibition of fibrinolytic activity as measured by increased tPAI-1 levels is in line with our results [5]. Elevated tPAI-1 levels have been associated with increased risk of cardiovascular events not only in patients with coronary artery disease but also in healthy participants [6, 7]. Moreover, it is well documented that high doses of any kind of alcohol beverage increases the risk of thromboembolic events, such as myocardial infarction, stroke, and peripheral arterial disease [1, 8].
In conclusion, drinking a high dose of red wine leads to acute inhibition of fibrinolysis in healthy men. This may partly explain the increased risk of cardiovascular mortality among binge drinkers. It seems that although red wine has some beneficial properties when consumed in moderate doses, its cardioprotective range is narrow, and in heavy doses may be in fact more hazardous than cognac.
Footnotes
Acknowledgements
The study is part of the project ‘Molecular Imaging in Cardiovascular and Metabolic Research’, which belongs to the Centre of Excellence programme of the Academy of Finland. This study was also supported by the Turku University Hospital Research Foundation, the Tampere University Hospital Medical Fund, the Turku University Foundation, the Emil Aaltonen Foundation, the Orionpharma Research Foundation, the Instrumentarium Research Foundation, the Finnish Foundation for Cardiovascular Research, and the Paulo Foundation.
Conflicts of interest: none declared.