Serenella Rotondo

Meta-Analysis of Wine and Beer Consumption in Relation to Vascular Risk

Background—Many epidemiological studies have evaluated whether different alcoholic beverages protect against cardiovascular disease. We performed a meta-analysis of 26 studies on the relationship between wine or beer consumption and vascular risk. Methods and Results—General variance-based method and fitting models were applied to pooled data derived from 26 studies that gave a quantitative estimation of the vascular risk associated with either beverage consumption. From 13 studies involving 209 418 persons, the relative risk of vascular disease associated with wine intake was 0.68 (95% confidence interval, 0.59 to 0.77) relative to nondrinkers. There was strong evidence from 10 studies involving 176 042 persons to support a J-shaped relationship between different amounts of wine intake and vascular risk. A statistically significant inverse association was found up to a daily intake of 150 mL of wine. The overall relative risk of moderate beer consumption, which was measured in 15 studies involving 208 036 persons, was 0.78 (95% confidence interval, 0.70 to 0.86). However, no significant relationship between different amounts of beer intake and vascular risk was found after meta-analyzing 7 studies involving 136 382 persons. Conclusions—These findings show evidence of a significant inverse association between light-to-moderate wine consumption and vascular risk. A similar, although smaller association was also apparent in beer consumption studies. The latter finding, however, is difficult to interpret because no meaningful relationship could be found between different amounts of beer intake and vascular risk.

Effect of trans‐resveratrol, a natural polyphenolic compound, on human polymorphonuclear leukocyte function

Polymorphonuclear leukocytes (PMN) may contribute to the pathogenesis of acute coronary heart disease (CHD). Epidemiological and laboratory evidence suggests that red wine, by virtue of its polyphenolic constituents, may be more effective than other alcoholic beverages in reducing the risk of CHD mortality. The aim of the present study was to investigate the effects of trans‐resveratrol (3,4′,5‐trihydroxy‐trans‐stilbene), a polyphenol present in most red wines, on functional and biochemical responses of PMN, upon in vitro activation. trans‐Resveratrol exerted a strong inhibitory effect on reactive oxygen species produced by PMN stimulated with 1 μM formyl methionyl leucyl phenylalamine (fMLP) (IC50 1.3±0.13 μM, mean±s.e.mean), as evaluated by luminol‐amplified chemiluminescence. trans‐Resveratrol prevented the release of elastase and β‐glucuronidase by PMN stimulated with the receptor agonists fMLP (1 μM, IC50 18.4±1.8 and 31±1.8 μM), and C5a (0.1 μM, IC50 41.6±3.5 and 42±8.3 μM), and also inhibited elastase and β‐glucuronidase secretion (IC50 37.7±7 and 25.4±2.2 μM) and production of 5‐lipoxygenase metabolites leukotriene B4 (LTB4), 6‐trans‐LTB4 and 12‐trans‐epi‐LTB4 (IC50 48±7 μM) by PMN stimulated with the calcium ionophore A23187 (5 μM). trans‐Resveratrol significantly reduced the expression and activation of the β2 integrin MAC‐1 on PMN surface following stimulation, as revealed by FACS analysis of the binding of an anti‐MAC‐1 monoclonal antibody (MoAb) and of the CBRM1/5 MoAb, recognizing an activation‐dependent epitope on MAC‐1. Consistently, PMN homotypic aggregation and formation of mixed cell‐conjugates between PMN and thrombin‐stimulated fixed platelets in a dynamic system were also prevented by trans‐resveratrol. These results, indicating that trans‐resveratrol interferes with the release of inflammatory mediators by activated PMN and down‐regulates adhesion‐dependent thrombogenic PMN functions, may provide some biological plausibility to the protective effect of red wine consumption against CHD.

Prevention of cardiovascular disease in type-2 diabetes: How to improve the clinical efficacy of aspirin

Atherosclerotic cardiovascular disease and its thrombotic complications are the principal causes of morbidity and mortality in patients with type-2 diabetes. Aspirin reduces the risk of thrombotic events in a broad range of patients with vascular disease and, in selected individuals, is beneficial for primary prevention. Although recommended by existing guidelines, in secondary and in primary prevention trials the clinical efficacy of low-dose aspirin in patients with diabetes appears to be substantially lower than in individuals without diabetes. In this review, we discuss possible mechanisms that may contribute to reduce the antithrombotic activity of aspirin in diabetes. We also discuss adjuvant therapies used in diabetic patients that may potentially improve the antithrombotic efficacy of aspirin.

Antithrombotic Effect of Polyphenols in Experimental Models

Abstract: Epidemiological studies have suggested that cardiovascular disease can be decreased by moderate wine consumption, but an overall quantitative estimation of the relationship between wine intake and vascular risk is lacking. A meta‐analysis was therefore performed on 19 studies selected on the basis of the availability of specific information on the cardiovascular relative risk (RR) associated with wine consumption. A significant risk reduction (RR: 0.66, 95% CI 0.57‐0.75) was associated with moderate (1‐2 drinks or 150‐300 mL/d) versus no wine consumption. In five studies which excluded ex‐drinkers as reference group, the overall RR associated with wine consumption was 0.61 (95% CI 0.57‐0.75). A dose‐response relation between wine intake and vascular risk resulted in a J‐shaped curve, with a significant risk reduction at about 300 mL/d (trend analysis p= 0.032). Two studies were also performed to investigate the effects of wine polyphenols on experimental thrombosis in rats. Supplementation for 10 days with alcohol‐free red wine—but not white wine or alcohol—induced a significant reduction of stasis‐induced venous thrombosis, an effect blunted by NO synthase inhibitor L‐NAME. In rats with diet‐induced hyperlipidemia, alcohol‐free red wine supplementation significantly delayed the thrombotic occlusion of an artificial prosthesis inserted into the abdominal aorta, but did not affect the increased cholesterol and triglyceride levels. TRAP values were significantly higher in animals receiving alcohol‐free wine. Altogether these experimental data support an antithrombotic role of polyphenols in the reduced vascular risk associated with moderate wine consumption in man, as shown by our epidemiological studies.

Licofelone, a dual lipoxygenase–cyclooxygenase inhibitor, downregulates polymorphonuclear leukocyte and platelet function

Polymorphonuclear leukocytes are strongly implicated in the pathogenesis of inflammatory disease. Polymorphonuclear leukocyte recruitment at sites of inflammation, mainly sustained by the beta2-integrins, is followed by the synthesis and release of inflammatory mediators, such as leukotrienes, proteolytic enzymes and reactive oxygen species. Functional and metabolic interactions between polymorphonuclear leukocytes and platelets can contribute to and exacerbate the process. The effects of the dual 5-lipoxygenase and cyclooxygenase inhibitor licofelone ([2,2-dimethyl-6-(4-chlorophenyl)-7-phenyl-2,3-dihydro-1H-pyrrolizine-5-yl]-acetic acid) were studied on arachidonic acid transcellular metabolism occurring between polymorphonuclear leukocytes and platelets. The formation of leukotriene C(4), a leukotriene A(4)-derived metabolite, by mixed polymorphonuclear leukocyte/platelet suspensions stimulated with 10 microM A23187 was inhibited by licofelone with an IC(50) of 3.8 +/- 0.07 microM. The formation of 5,12-di-hydroxy-eicosatetraenoic acid (HETE) was abolished at concentrations > or = 10 microM. Licofelone also inhibited the generation of reactive oxygen species by polymorphonuclear leukocytes stimulated with 1 microM n-formyl-methionyl-leucyl-phenylalanine (fMLP), 10 nM complement fraction 5a (C5a) and 1 microM platelet activating factor (PAF) with IC(50)s of 24.4 +/- 0.6, 11.0 +/- 1.5 and 11.7 +/-1.2 microM; elastase release induced by the three agonists was inhibited with IC(50)s of 12.2 +/- 2.2, 23.5 +/- 8 and 2.6 +/- 1 microM, respectively. Homotypic polymorphonuclear leukocyte aggregation induced by fMLP, C5A and PAF was inhibited by licofelone with IC(50)s of 23.7 +/- 4.8, 15.6 +/- 3.4 and 15.4 +/- 4 microM, respectively. The present study extends the anti-lipoxygenase and anti-cyclooxygenase activities of licofelone to the production of arachidonic acid metabolites generated as a consequence of polymorphonuclear leukocyte-platelet transcellular metabolism and to polymorphonuclear leukocyte responses relevant to the pathogenesis of inflammation. The coexistence within the same molecule of a wide spectrum of anti-inflammatory properties is of interest.

A meta-analysis of studies on wine and beer and cardiovascular disease

Many epidemiologic studies have evaluated whether different alcoholic beverages protect against cardiovascular disease. We performed a meta-analysis of 26 studies on relationship between wine or beer consumption and vascular risk. General variance-based method and fitting models were applied to pooled data derived from 26 studies that gave quantitative estimation of the vascular risk associated with either beverage consumption. From 13 studies involving 209,418 persons, the relative risk of vascular disease associated with wine intake was 0.68 (95% CI: 0.59-0.77) relative to nondrinkers. There was strong evidence from 10 studies involving 176,042 persons to support a J-shaped relationship between different amounts of wine intake and vascular risk. A statistically significant inverse association was found up to a daily intake of 150 ml of wine. The overall relative risk of moderate beer consumption, which was measured in 15 studies involving 208,036 persons, was 0.78 (95% CI: 0.70-0.86). However, no significant relationship between different amounts of beer intake and vascular risk was found after meta-analyzing 7 studies involving 136,382 persons. These findings show evidence of a significant inverse association between light to moderate wine consumption and vascular risk. A similar, although smaller association was also apparent in beer consumption studies. The latter finding, however, is difficult to interpret because no meaningful relationship could be found between different amounts of beer intake and vascular risk.

Inhibition by soya isoflavones of human polymorphonuclear leukocyte function: possible relevance for the beneficial effects of soya intake.

Lower CVD incidence is reported in Asian populations consuming soya-containing food. As polymorphonuclear leukocytes (PMN) are involved in the risk of CVD, we investigated the modulatory effect of soya isoflavones on several PMN functions and their molecular mechanisms in vitro. PMN, isolated from blood from healthy subjects, were tested upon activation with 1 microm- n-formyl-methyl-leucyl-phenylalanine (fMLP) for superoxide anion production (ferric cytochrome c reduction) and released elastase (chromogenic test). PMN homotypic aggregates stimulated by fMLP or P-selectin in dynamic conditions were detected by optical microscopy. PMN, mixed with thrombin-activated, washed platelets, formed cell aggregates, measured by flow cytometry. Phosphorylation of Pyk2, a focal adhesion kinase, was studied by immunoprecipitation and immunoblotting with specific antibodies. Genistein, daidzein and equol inhibited superoxide anion production (IC50 0.25 (sem 0.1), 21.0 (sem 4.2) and 13.0 (sem 2.8) microm, respectively); the release of elastase was prevented by genistein (IC50 63 (sem 17) microm). PMN homotypic aggregates, stimulated by fMLP, were significantly reduced (24 (sem 12) and 51 (sem 14) % of control) by 100 microm genistein and equol. P-selectin-induced aggregates were reduced to 19 (sem 6), 44 (sem 10) and 28 (sem 9) % of control by 100 microm genistein, daidzein and equol, respectively. Genistein, daidzein and equol also significantly reduced mixed platelet-PMN aggregates (IC50 4.0 (sem 0.9), 57 (sem 6) and 66 (sem 23) microm, respectively). In PMN challenged by fMLP or P-selectin, activation of Pyk2 was prevented by isoflavones. The cardioprotective effect of soya-containing food might be linked to reduction of PMN activation and PMN-platelet interaction, novel targets for the biological effects of soya isoflavones.

Ticlopidine Does Not Reduce In Vivo Platelet Thromboxane Biosynthesis and Metabolism in Diabetic Patients

Diabetic patients are at higher risk of development of cardiovascular complications than the general population. The role of platelets in the pathogenesis of these complications is still controversial, it being difficult to ascertain whether altered platelet function is a cause or consequence of vascular complications of diabetes. Measurement of urinary 11-dehydro-thromboxane has been proposed as a reliable index of in vivo platelet activation and has been reported to be significantly higher in non insulin-dependent diabetic patients with micro- or macrovascular complications. We therefore studied the effect of ticlopidine, an antiplatelet drug acting through mechanisms different from cyclo-oxygenase inhibition, on urinary 11-dehydro-TXB(2) excretion in diabetic patients with macrovascular complications. The results indicate that urinary excretion of 11-dehydro-TXB(2) after ticlopidine treatment is not different from pre-treatment values, suggesting that the chosen parameter might not be reliable for monitoring the antiplatelet activity of ticlopidine and possibly of other drugs which do not directly affect arachidonic acid metabolism.

The endoperoxides/TxA2 analogue, U46619, inhibits human polymorphonuclear leukocyte function.

The effects of the stable analogue of TxA2, U46619, on polymorphonuclear leukocyte (PMN) function were investigated. U46619, at micromolar concentrations, inhibited fMLP‐stimulated aggregation, β‐glucuronidase release, and superoxide production. fMLP‐induced LTB4 synthesis was also inhibited. U46619 did not modify intracellular Ca2+ increase induced by fMLP in Fura‐2‐loaded PMN, suggesting that early events of cell activation were not involved. In fact, U46619 also inhibited aggregation, β‐glucuronidase release, superoxide anion and LTB4 production induced by the calcium ionophore A23187. By comparison with the specific 5‐lipoxygenase inhibitor, L‐663,536, which prevented LTB4 synthesis without affecting degranulation, we excluded the impairment of PMN function by U46619 as a consequence of the reduction of this endogenous agonist. TLC separation of lipid extracts from [3H]‐AA‐loaded PMN, stimulated by A23187, showed significant reduction of the radioactivity associated with authentic free AA, suggesting that U46619 could interfere with mechanisms regulating AA release from membrane phospholipids. This suggestion is also supported by the observation that manoalide, a standard inhibitor of phospholipase A2, similarly to U46619, inhibits β‐glucuronidase release from stimulated PMN. Prostaglandin endoperoxides, produced by cells participating in inflammatory reactions, might therefore play a role in modulating PMN activities. J. Leukoc. Biol. 57: 72–79; 1995.

Antiplatelet activity of 2-(6-carboxyhexyl)-3-n-hexylcyclohexylamine (IBI P-05006), a thromboxane receptor antagonist

Abstract 2-(6-Carboxyhexyl)-3-n-hexylcyclohexylamine (IBI P-05006) is a molecule with cytoprotective and antisecretory actions. Since its chemical structure resembles that of the prostanoid nucleus of thromboxane (Tx) A2, the possibility of antiplatelet activity was investigated. The aims of our study were to evaluate a possible inhibitory effect of IBI P-05006 on in vitro platelet aggregation induced by different stimuli and to characterize its mechanism of action. For this purpose, selected mechanisms of platelet function were evaluated. IBI P-05006 inhibited platelet aggregation induced by arachidonic acid or U46619, at concentrations ranging from 0.4 to 4 μM, dose dependently, without interfering with arachidonic acid metabolism. The possibility that the compound acts through adenylate cyclase stimulation was ruled out by measurement of cAMP levels, which were not increased. Receptor binding studies indicated competitive inhibition by IBI P-05006 of the binding of the ligand [3H]SQ29548 to the TxA2/(prostaglandin) PG-cyclic endoperoxides receptor with IC50 in the range of concentrations active on platelet aggregation. In conclusion, IBI P-05006 inhibits in vitro human platelet aggregation through a specific mechanism of competition for the TxA2/PG-cyclic endoperoxides receptor.