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Quinones M.,Complutense University of Madrid | Sanchez D.,Complutense University of Madrid | Muguerza B.,Natraceutical Group | Miguel M.,Institute Investigacion En Ciencias Of La Alimentacion Cial | Aleixandre A.,Complutense University of Madrid
Food Research International | Year: 2011

We investigate the mechanisms involved in the long-term antihypertensive effect of a polyphenol-rich cocoa powder, named CocoanOX® (CCX), in spontaneously hypertensive rats (SHR). We have carried out two different batches of experiments. For the first batch of experiments, forty 3 week-old male SHR were randomly divided with ad libitum intake into four groups of 10 animals, that respectively received the following drinking fluids up to the 20th week of life (treatment period): tap water (control), CCX 100 mg/kg/day, CCX 200 mg/kg/day and CCX 400 mg/kg/day. Five 20 week-old rats of each group were sacrificed by decapitation. From the 20th to 24th week of life all the remaining animals were given tap water (follow-up period), and all of them were sacrificed at the end of the follow-up period. Plasma malondialdehyde (MDA), reduced glutathione in the liver, plasma and aorta angiotensin converting enzyme (ACE) activity and plasma angiotensin II were determined in all the sacrificed SHR that were included in this batch of experiments. Plasma MDA decreased and liver reduced glutathione increased in the 20 week-old CCX treated SHR. These effects were not observed in the rats that were sacrificed after the follow-up period. CCX treatment did not modify aorta ACE activity, but the activity of ACE and the levels of angiotensin II increased in the plasma of the SHR treated with the highest dose of CCX. ACE activity returned to basal values in the SHR that were sacrificed after the follow-up period. However, angiotensin II levels were slightly higher after withdrawal of CCX.For the second batch of experiments we used aorta rings obtained from untreated SHR, and we evaluated the relaxation caused by CCX in different aorta preparations. CCX relaxed the intact aorta preparations but this cocoa did not relax the endothelium-denuded aorta rings from the untreated SHR. l-NAME, but not indomethacin, inhibited the relaxation caused by CCX in the SHR aorta rings. We postulate that the antihypertensive effect of CCX might be mediated by an improvement of endothelial release of nitric oxide and by a reduction of oxidative stress. The inhibition of ACE could be implicated in the antihypertensive effect of CCX. © 2010 Elsevier Ltd.


Sanchez D.,Complutense University of Madrid | Quinones M.,Complutense University of Madrid | Moulay L.,Natraceutical Group | Muguerza B.,Natraceutical Group | And 3 more authors.
Journal of Agricultural and Food Chemistry | Year: 2010

The effect produced by long-term intake of a soluble cocoa fiber product (SCFP) on the development of hypertension of spontaneously hypertensive rats (SHR) was evaluated. Twenty male 3-week-old SHR were divided into two groups of 10 animals that drank either tap water (control) or a solution of SCFP (0.75 g/day SCFP) until the 20th week of life. Five 20-week-old rats of each group were sacrificed. Tap water as drinking fluid was given to all the animals from the 20th to 24th week of life. The 24-week-old rats were also sacrificed. Body weight, liquid and dry food intake, and arterial blood pressure (tail cuff) were recorded weekly. Malondialdehyde (MDA), glucose and angiotensin converting enzyme (ACE) activity in the plasma from the sacrificed rats were also obtained, and we evaluated the relaxation caused by acetylcholine in the aorta from these animals. SCFP attenuated the development of hypertension in SHR; however, the withdrawal of SCFP caused an increase in blood pressure in the rats. Body weight gain was slower in the group treated with SCFP. SCFP increased liquid intake but decreased dry food intake in the rats. SCFP decreased plasma MDA concentrations and slightly decreased plasma ACE activity, but no differences were observed in plasma glucose and in the aorta responses to acetylcholine in both groups of 20-week-old SHR. We have demonstrated the antihypertensive and antioxidant properties of SCFP. The control of body weight and the control of increased angiotensin Il may be involved in the antihypertensive effect of this product. © 2010 American Chemical Society.


Sanchez D.,Complutense University of Madrid | Moulay L.,Natraceutical Group | Muguerza B.,Natraceutical Group | Quinones M.,Complutense University of Madrid | And 3 more authors.
Journal of Medicinal Food | Year: 2010

The effects of a soluble cocoa fiber (SCF) were studied in Zucker fatty rats. Two groups of Zucker fatty rats were fed the following diets: standard diet and 5% SCF-enriched diet. A group of Zucker lean rats fed the standard diet was used for results comparison with obese Zucker animals. Solid and liquid intakes, body weight, plasma glucose, lipid profile, and systolic (SBP) and diastolic (DBP) blood pressure were recorded weekly. At the end of the experimental period insulin was determined, and fat apparent digestibility (FAD) and insulin resistance were calculated. The Zucker fatty rats fed 5% SCF-enriched diet showed less weight gain and food intake than those fed the standard diet. The group fed the fiber-enriched diet showed lower values of the total cholesterol/high-density lipoprotein cholesterol ratio and triglyceride levels than the standard group. FAD was also lower in the fiber group. Both SBP and DBP were decreased. In addition, SCF reduced plasma glucose and insulin, and as a consequence the insulin resistance was also decreased. Our data demonstrate that SCF resulted in an improvement of the studied risk factors associated with cardiometabolic disorders. © 2010, Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition.


Quinones M.,Complutense University of Madrid | Miguel M.,Institute Investigacion En Ciencias Of Alimentacion | Muguerza B.,Natraceutical Group | Muguerza B.,Rovira i Virgili University | Aleixandre A.,Complutense University of Madrid
Food and Function | Year: 2011

In this study, we evaluated the short-term effect of a cocoa polyphenol extract (CPE), in spontaneously hypertensive rats (SHR). Male 17-22-week-old SHR were administered by intragastric gavage water, 50 mg kg-1 Captopril or CPE at different doses (13, 26, 80 and 160 mg kg-1). The systolic blood pressure (SBP) and diastolic blood pressure (DBP) were recorded by the tail cuff method before the administration and also 2, 4, 6, 8, 24, 48 and 72 h post-administration. Highly significant decreases in the SBP and in the DBP were observed when captopril or CPE was administered to SHR. The cocoa extract produced a dose dependent effect in the SBP of the SHR up to the dose of 80 mg kg-1. Nevertheless this dose of CPE did not decrease the arterial blood pressure in the normotensive Wistar Kyoto rats. The decrease in the SBP caused by 80 mg kg-1 of CPE in the SHR (-39.1 ± 3.7 mm Hg) was maximum 6 h post-administration, and the initial values of SBP were recovered 72 h post-administration of this extract. Paradoxically, 160 mg kg-1 of the cocoa extract caused a decreased antihypertensive effect than lower doses of CPE. In addition, the decrease in DBP was always more accentuated when the dose of CPE administered was lower. Our results suggest that CPE may be used as a functional food ingredient with beneficial effects for controlling arterial blood pressure. This journal is © 2011 The Royal Society of Chemistry.


Quinones M.,Complutense University of Madrid | Sanchez D.,Complutense University of Madrid | Muguerza B.,Natraceutical Group | Moulay L.,Natraceutical Group | And 4 more authors.
Food Chemistry | Year: 2010

We have evaluated the effect of the long-term intake of a cocoa powder, with high concentration of polyphenols, named CocoanOX (CCX), on the development of hypertension of spontaneously hypertensive rats (SHR). Systolic blood pressure was measured weekly in the rats, from the 6th to 24th week of life, by the tail cuff method. The development of hypertension was attenuated in the groups treated with captopril or CCX. The antihypertensive effect was more accentuated in the group treated with captopril, and it was paradoxically more accentuated in the group treated with the lowest dose of CCX than in the other CCX groups. The arterial blood pressure increased in the treated SHR when the corresponding antihypertensive treatment was removed. Both, CCX and the standard cocoa, improved the aorta endothelial function in the SHR. In conclusion, CCX could be used as a functional food ingredient with antihypertensive activity. © 2010 Elsevier Ltd. All rights reserved.


Schinella G.,National University of La Plata | Mosca S.,National University of La Plata | Cienfuegos-Jovellanos E.,Natraceutical Group | Pasamar M.A.,Natraceutical Group | And 3 more authors.
Food Research International | Year: 2010

Fermentation and roasting are the main causes of polyphenol degradation during the process for obtaining cocoa products. In the present study, a process for obtaining polyphenol-rich cocoa products on an industrial scale is described. The process avoids the fermentation and roasting steps and includes a step for the inactivation of the enzyme Polyphenol Oxidase (PPO), which helps preserve the polyphenol content present in the raw cocoa bean. In addition, our study evaluates the antioxidant capacity and characterizes the flavonoid profile of the polyphenol-rich cocoa products obtained from the natural polyphenol-rich cocoa cake. Using different protocols, we have obtained three cocoa extracts with high polyphenol content, namely extracts A (167mg/g), B (374mg/g) and C (787mg/g). The scavenging capacity of the extracts was measured as their ability to bleach the stable radicals DPPH and ABTS+ while their antioxidant effect was evaluated with the FRAP assay. The results for A, B and C in the DPPH test expressed as Trolox equivalent (μmol)/mg dry weight of extract were 0.2, 1.4 and 3.0, respectively; in the ABTS test the results were 1.0, 4.7 and 9.8. The antioxidant capacity expressed as ascorbic acid equivalent (μmol)/mg dry weight of each product were 17.2, 76.1 and 207.7, respectively. The scavenging properties of cocoa powder against the superoxide anion, H2O2, HClO, and peroxynitrite were also determined. The IC50 (μg/mL) values in the hypoxanthine/xanthine oxidase test were 77.5, 12.3 and 10.3, for A, B and C, respectively, while as an HOCl scavenger the IC50 (μg/mL) values were 225.4, 73.2 and 21.5. As a peroxynitrite anion scavenger, only extract C had a relevant effect, with IC50 (μg/mL) values of 76.1 or 110.0 in the absence or presence of bicarbonate. None of the extracts tested showed activity in the hydrogen peroxide test, but B and C significantly increased the deoxyribose degradation in the absence of ascorbate. Likewise, none of the extracts inhibited the ferrous or copper chelating activity at 100μg/mL, but they inhibited the lipid peroxidation in brain homogenates and human plasma through non-enzymatic generation systems, with extract C giving the best IC50 (μg/mL) values: 17.4 and 8.1 against lipid peroxidation in brain homogenates and human plasma, respectively. In conclusion, if the extractive protocol is well characterized, defined and optimized, cocoa could constitute a source of polyphenols for enriching foods, nutraceuticals and alimentary supplements. © 2010 Elsevier Ltd.


Andujar I.,University of Valencia | Recio M.C.,University of Valencia | Giner R.M.,University of Valencia | Cienfuegos-Jovellanos E.,Natraceutical Group | And 3 more authors.
Journal of Agricultural and Food Chemistry | Year: 2011

We studied a polyphenol-enriched cocoa extract (PCE) with epicatechin, procyanidin B2, catechin, and procyanidin B1 as the major phenolics for its anti-inflammatory properties against dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice. PCE reduced colon damage, with significant reductions in both the extent and the severity of the inflammation as well as in crypt damage and leukocyte infiltration in the mucosa. Analysis ex vivo showed clear decreases in the production of nitric oxide, cyclooxygenase-2, pSTAT-3, and pSTAT1α, with NF-B p65 production being slightly reduced. Moreover, NF-B activation was reduced in RAW 264.7 cells in vitro. In conclusion, the inhibitory effect of PCE on acute UC induced by DSS in mice was attenuated by oral administration of PCE obtained from cocoa. This effect is principally due to the inhibition of transcription factors STAT1 and STAT3 in intestinal cells, with NF-B inhibition also being implicated. © 2011 American Chemical Society.

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