Cacao Polyphenol Research Articles

Dose-dependent Effects of Cacao Polyphenol Intake on Lipid Metabolism in Rats.

Cacao polyphenols (CPs) are known to suppress the oxidation of low-density lipoprotein and cholesterol absorption. Herein, we examined the impact of CP on the lipid metabolism in rats fed CP-rich chocolate, by analyzing liver weight and histology, via hematoxylin-eosin staining. The high-CP group had significantly lighter livers than the CP-free group. Histologically, the high-CP group showed significantly lower liver fat accumulation than the CP-free group. These results suggest that CPs prevent liver fat accumulation, being potentially useful against obesity and related diseases.

Clovamide, a Hydroxycinnamic Acid Amide, Is a Resistance Factor Against Phytophthora spp. in Theobroma cacao.

The hydroxycinnamic acid amides (HCAAs) are a diverse group of plant-specialized phenylpropanoid metabolites distributed widely in the plant kingdom and are known to be involved in tolerance to abiotic and biotic stress. The HCAA clovamide is reported in a small number of distantly related species. To explore the contribution of specialized metabolites to disease resistance in cacao (Theobroma cacao L., chocolate tree), we performed untargeted metabolomics using liquid chromatography – tandem mass spectrometry (LC-MS/MS) and compared the basal metabolite profiles in leaves of two cacao genotypes with contrasting levels of susceptibility to Phytophthora spp. Leaves of the tolerant genotype ‘Scavina 6’ (‘Sca6’) were found to accumulate dramatically higher levels of clovamide and several other HCAAs compared to the susceptible ‘Imperial College Selection 1’ (‘ICS1’). Clovamide was the most abundant metabolite in ‘Sca6’ leaf extracts based on MS signal, and was up to 58-fold higher in ‘Sca6’ than in ‘ICS1’. In vitro assays demonstrated that clovamide inhibits growth of three pathogens of cacao in the genus Phytophthora, is a substrate for cacao polyphenol oxidase, and is a contributor to enzymatic browning. Furthermore, clovamide inhibited proteinase and pectinase in vitro, activities associated with defense in plant-pathogen interactions. Fruit epidermal peels from both genotypes contained substantial amounts of clovamide, but two sulfated HCAAs were present at high abundance exclusively in ‘Sca6’ suggesting a potential functional role of these compounds. The potential to breed cacao with increased HCAAs for improved agricultural performance is discussed.

Abstract WP465: Effect of Chocolate Containing 86% Cacao and Much Polyphenol to Improve Level of Alertness in Acute Stroke Patients

Introduction: Level of alertness in acute stroke patients is often deteriorated during hospitalization, even though they can orally eat food on admission and their stroke is not severe. Then, it is difficult to improve their level of alertness. If their level of alertness is not improved, their nutritional status is probably deteriorated. Hypothesis: Dark chocolate 5g tablet containing 86% cacao and 147mg polyphenol can improve level of alertness and prevent deterioration of nutritional status. Methods: We included acute stroke patients for retrospective analysis who 1) were admitted between August 2017 and July 2018, 2) presented alert consciousness on admission and orally eat food then, 3) underwent blood examination on admission, the 3rd day and 7th day, 4) presented mild deterioration of level of alertness during hospitalization and 5) took dark chocolate 5g containing 86% cacao and 147mg polyphenol twice a day. We evaluated patients’ features, Glasgow Coma Scale (GCS) as level of consciousness, albumin (Alb), prealbumin (PreAlb), high-sensitivity C-reactive protein (hs-CRP) on admission, on the initial chocolate day and on the 7th day. Results: Eighteen patients met our inclusive criteria and were analyzed. The median age, GCS score, body mass index, blood glucose (BG), hs-CRP, Alb and PreAlb were 85 years, 14, 21.8 kg/m2, 119 mg/dl, 0.1165 mg/dl, 3.9 g/dl and 20.4 mg/dl, respectively. They started to eat 86% cacao chocolate 5g supplementation twice a day median on the 3rd day. On the 3rd day when chocolate was started, their median GCS score, Alb and PreAlb decreased to13 (p<0.0001), 3.45 mg/dl (p<0.0001) and 19.1 mg/dl (p<0.001), respectively, and their median hs-CRP increased to 0.4325 mg/dl (p<0.05). On the 7th day, their median GCS increased to 14 (0.0001), median Alb, PreAlb and hs-CRP levels were 3.45 mg/dl (ns), 16.5 g/dl (ns) and 0.506 mg/dl (ns). Conclusion: Dark chocolate containing 86% cacao and 147mg polyphenol probably improved level of alertness and prevented further deterioration of nutritional status in patients who could orally eat food.

Effects of regular high-cocoa chocolate intake on arterial stiffness and metabolic characteristics during exercise.

The aim of this study was to examine the effects of regular high-cocoa chocolate consumption on arterial stiffness and fat oxidation during light- to moderate-intensity exercise. This randomized, controlled, parallel-group intervention study included 32 Japanese college students (mean age, 20.7 ± 0.3 y; men, n = 24; women, n = 8) who were assigned to either control or intervention groups (n = 16 each). The control group did not alter their habitual diet or physical activity throughout the study period. The intervention group consumed 20 g/d (508 mg of cacao polyphenol) of high-cocoa chocolate for 4 wk. Blood pressure, heart-ankle pulse wave velocity, cardio-ankle vascular index, body composition, and metabolic characteristics during exercise at 50% maximal oxygen uptake level were assessed before and after the intervention. Four weeks of high-cocoa chocolate ingestion significantly reduced heart-ankle pulse wave velocity and cardio-ankle vascular index (%change, intervention versus control: -2.3 ± 0.9% versus 0.9 ± 0.9%, and -4.8 ± 1.8% versus 0.7 ± 1.3%, respectively; both P < 0.05). However, blood pressure, weight, body mass index, body fat, waist circumference, and metabolic characteristics during exercise such as respiratory exchange ratio did not significantly change in either group. Four weeks of regular high-cocoa chocolate consumption reduced arterial stiffness after considering blood pressure in healthy young men and women. However, the habitual consumption of high-cocoa chocolate for 4 wk did not affect metabolic characteristics during light- to moderate-intensity exercise and body composition.

Comparison of Polyphenol Concentration and Composition between Genetically Diverse Cacao (Theobroma cacao L.) Accessions Selected for High Yield

There is abundant evidence that consumption of cacao and dark chocolate promotes human health and that the main cacao components contributing positive health effects are polyphenols. The polyphenols in cacao bean constitute 12–18% dry weight of the whole bean and are predominantly catechins (37% w/w), procyanidins (58% w/w) and anthocyanins (4% w/w). Variation in cacao polyphenol composition and concentration can result from differences in genetic backgroud, agro‐environment and cultural practices. It is therefore important to consider several factors when selecting cacao seed (beans) for research investigating the mechanisms underlying its health benefits. In this study, dried unfermented seeds of five different cacao clones (TARS‐9, TARS‐14, TARS‐27, TARS‐30, TARS‐31) were obtained from USDA‐ARS Tropical Agriculture Research Station (TARS) located in Mayaguez, Puerto Rico. These clones were developed through controlled‐pollinated hybridization for improved agronomic performance, specifically high yield. Prior sensory evaluation of 65% chocolate made from these clones revealed that each conferred distinct profiles in bitterness. We thus hypothesized that differences in polyphenol profiles would exist between these clones. To test this hypothesis, total polyphenols, total flavan‐3ols, polyphenol extraction yield, and qualitative flavanol profile were compared between the five cacao clones. TARS‐27 had the lowest total polyphenol concentration (36 ± 1.2 mg gallic acid equivalents/g cacao bean) (mean ± STDEV), whereas TARS‐14 showed highest concentration (66 ± 4.7 mg gallic acid equivalents/g cacao bean). For total flavan‐3‐ols, a similar trend was observed; TARS‐27 showed the lowest concentration (42.3 ± 1.1 mg procyanidin B2 equivalents/g cacao bean) with TARS‐9 having the highest concentration (76.7 ± 4 mg procyanidin B2 equivalents/g cacao bean). Polyphenol extraction yield (cacao extract weight/cocoa bean weight) differed among clones ranging from 0.1–0.16. Qualitative flavanol profiles obtained using normal‐phase high‐performance liquid chromatography (HPLC) indicated that mean degree of polymerization (mDP) of polyphenols is likely similar between these clones. Further research is required to quantitatively determine mDP, and to identify and quantify individual polyphenols. In conclusion, these insights into the variability of polyphenol composition among genetically diverse cacao will help to inform targeted selection of cacao clones for future studies investigating the impact of cacao consumption on human health.Support or Funding InformationFunding was provided, in part, by the Virginia Agricultural Experiment Station and the Hatch Program of the National Institute of Food and Agriculture (NIFA), U.S. Department of Agriculture

Corrigendum: Cacao polyphenols ameliorate autoimmune myocarditis in mice.

Correction to: Hypertension Research (2016) 39, 203–209; doi:10.1038/hr.2015.136 After publication of this article, the authors noticed an error in ‘Cacao polyphenol administration’ in the Methods section. ‘The CPE group (n=12) received CPE in distilled water (1.2 mg kg−1 per day)’ was incorrect. The correct concentration of CPE was 1.

Cacao polyphenols ameliorate autoimmune myocarditis in mice.

Myocarditis is a clinically severe disease; however, no effective treatment has been established. The aim of this study was to determine whether cacao bean (Theobroma cacao) polyphenols ameliorate autoimmune myocarditis. We used an experimental autoimmune myocarditis (EAM) model in Balb/c mice. Mice with induced EAM were treated with a cacao polyphenol extract (CPE, n=12) or vehicle (n=12). On day 21, hearts were harvested and analyzed. Elevated heart weight to body weight and fibrotic area ratios as well as high cardiac cell infiltration were observed in the vehicle-treated EAM mice. However, these increases were significantly suppressed in the CPE-treated mice. Reverse transcriptase-PCR revealed that mRNA expressions of interleukin (Il)-1β, Il-6, E-selectin, vascular cell adhesion molecule-1 and collagen type 1 were lower in the CPE group compared with the vehicle group. The mRNA expressions of nicotinamide adenine dinucleotide phosphate-oxidase (Nox)2 and Nox4 were increased in the vehicle-treated EAM hearts, although CPE treatment did not significantly suppress the transcription levels. However, compared with vehicle treatment of EAM hearts, CPE treatment significantly suppressed hydrogen peroxide concentrations. Cardiac myeloperoxidase activity, the intensity of dihydroethidium staining and the phosphorylation of nuclear factor-κB p65 were also lower in the CPE group compared with the vehicle group. Our data suggest that CPE ameliorates EAM in mice. CPE is a promising dietary supplement to suppress cardiovascular inflammation and oxidative stress.

Carbon tetrachloride-induced hepatic and renal damages in rat: inhibitory effects of cacao polyphenol.

Here, we investigated the protective effect of cacao polyphenol extract (CPE) on carbon tetrachloride (CCl4)-induced hepato-renal oxidative stress in rats. Rats were administered CPE for 7 days and then received intraperitoneal injection of CCl4. Two hours after injection, we found that CCl4 treatment significantly increased biochemical injury markers, lipid peroxides (phosphatidylcholine hydroperoxide (PCOOH) and malondialdehyde (MDA)) and decreased glutathione peroxidase activity in kidney rather than liver, suggesting that kidney is more vulnerable to oxidative stress under the present experimental conditions. CPE supplementation significantly reduced these changes, indicating that this compound has antioxidant properties against CCl4-induced oxidative stress. An inhibitory effect of CPE on CCl4-induced CYP2E1 mRNA degradation may provide an explanation for CPE antioxidant property. Together, these results provide quantitative evidence of the in vivo antioxidant properties of CPE, especially in terms of PCOOH and MDA levels in the kidneys of CCl4-treated rats.

Effect of Theobroma cacao Polyphenol on Isoproterenol-Induced Myocardial Infarction in Wistar Rats

Evaluation of cardioprotective effects of Parkia biglobosa (Jacq. Benth) Mimosaceae stem barkAdi K., Metowogo K., Mouzou A., Lawson-Evi P., Eklu-Gadegbeku K., Agbonon A., Lamboni C., Essien K., Aklikokou K., Gbeassor M.

Theobromine enhances absorption of cacao polyphenol in rats.

Several concentrations of theobromine (TB) and (-)-epicatechin (EC) were coadministered to rats, and plasma EC and its metabolites were determined using ultra-high-performance liquid chromatography-tandem mass spectrometry. It has been demonstrated that TB increases the absorption of EC in a dose-dependent manner. Cocoa powder had a similar effect, and the mechanism involved is not thought to depend on tight junctions.

Oxidative Stress during Development of Alcoholic Fatty Liver: Therapeutic Potential of Cacao Polyphenol

The lipid and antioxidative/oxidative profiles of livers from rats fed an ethanol liquid diet for 8 weeks provided evidence for an involvement of oxidative stress (e.g., phospholipid peroxidation) in the development of alcoholic fatty liver (AFL), possibly in an early stage. Cacao polyphenol supplementation produced an ameliorating effect, and may help in AFL prevention.

Natural Ghana Cacao Powder의 Polyphenol 성분분석 및 피부개선효과 연구

가나는 카카오의 최대산지이며, 카카오는 식품으로 이용하는 초코렛 재료가 되는 원료이며 오래전서부터 항산화 효과등이 알려져서 다양한 식품에 사용되어 왔다. 또한 가나산 카카오는 다른지역 카카오에 비해 폴리페놀 함량이 높은 것으로 알려져 왔으며 알칼리 처리를 하지 않아 약산성의 자연그대로의 특징이 남아 있다. 이 천연 가나산 카카오의 폴리페놀 함량을 분석하고 이것을 이용한 화장품 팩제를 개발하여 피부자극실험과 피부개선효과를 연구하였다. 그 결과 가나산 카카오파우더에는 약 3.6%의 탄닌성분이 포함 되어 있는 것으로 나타났으며 pH는 5.6을 나타내었고 가나산 카카오 파우더를 15% 함유한 팩제의 실험결과 일차피부자극실험에서는 피부자극이 나타나지 않았으며 120분후의 피부개선효과를 본 실험에서는 피부 수분량은 20%증가, 피부수분증발량은 17% 감소 피부pH는 5.3수렴, 피부탄력도는 24% 증가하는 것으로 나타나 피부개선 효과가 나타난 것으로 연구되었다. Ghana is the country that produces world's biggest production of Cacao. Cacao is the main ingredient of chocolate, which has been widely used in a variety of food as its anti-oxidantal effect is well known to public. Moreover, Ghana-produced Cacao is known to have a bigger amount of polyphenol compared to the ones produced elsewhere, and as they are not processed with alkali Ghana-produced Cacoa is slightly acidic as it is. This project aimed at analysing this natural Ghana Cacao's polyphenol composition, developed cosmetic mask using it and don skin irritation tests in order to study skin improvement effect. As a result, it was found that Ghana-produced Cacao contains approximately 3.6% of tannin, showing 5.6 pH. From the result of the first skin irritation test, the result of experiment of the cometic masks which contain 15% of Ghana cacao powder showed that the irritation was not shown. After 120 minutes, in the experiment of skin improvement effect, it was proved to have skin improvement effect, appearing 20% increase in skin moisture, -17% decrease in skin moisture evaporation, convergence of 5.3pH, 24% increase in skin elasticity.

Cacao Polyphenol Extract Suppresses Transformation of an Aryl Hydrocarbon Receptor in C57BL/6 Mice

Dioxins enter the body through the diet and cause various toxicological effects through transformation of an aryl hydrocarbon receptor (AhR). Plant extracts and phytochemicals including flavonoids are reported to suppress this transformation. This paper investigates the suppression by a cacao polyphenol extract (CPE) of AhR transformation in vivo. The CPE was administered orally to C57BL/6 mice at 100 mg/kg of body weight, followed 1 h later by 3-methylcholanthrene (MC), an AhR agonist, injected intraperitoneally at 10 mg/kg of body weight. CPE suppressed the MC-induced transformation to the control level by inhibiting the formation of a heterodimer between AhR and an aryl hydrocarbon receptor nuclear translocator in the liver at 3 h postadministration. It also suppressed MC-induced cytochrome P4501A1 expression and NAD(P)H:quinone-oxidoreductase activity, whereas it increased glutathione S-transferase activity at 25 h. CPE constituents and their metabolites might contribute, at least in part, to the suppression of AhR transformation. The results indicate that the intake of CPE suppressed the toxicological effects of dioxins in the body.

Acute effect of oral flavonoid-rich dark chocolate intake on coronary circulation, as compared with non-flavonoid white chocolate, by transthoracic Doppler echocardiography in healthy adults

Purpose To assess the effects of the oral intake of flavonoid-rich dark chocolate on coronary circulation, we measured coronary flow velocity reserve (CFVR) by noninvasive transthoracic Doppler echocardiography (TTDE) in healthy adult subjects. Materials and methods The study was a randomized, single-blind design conducted for 2 weeks in 39 healthy men (mean age 29.7 ±3.9 years, range 23–40 years). Subjects were randomly assigned a daily intake of either flavonoid-rich dark chocolate (Meiji Black Chocolate 45 g, Meiji Seika kaisya Ltd, including cacao polyphenol 550 mg/day, 200 kcal) or non-flavonoid white chocolate (Meiji White Chocolate 35 g, Meiji Seika kaisya Ltd, including cacao polyphenol 0 mg/day, 140 kcal) as a control. CFVR was recorded by TTDE, and assessed before and after 2 weeks of intake. At the same time, we also assessed serum asymmetric dimethylarginine, 8-isoprostanes, and malondialdehyde-modified low-density lipoprotein (MDA-LDL) as markers of oxidative stress. Results Flavonoid-rich dark chocolate consumption significantly improved CFVR (3.38 ± 0.49 before intake, 4.28 ± 0.85 after intake; p < 0.01), whereas non-flavonoid white chocolate consumption did not (3.28 ± 0.49 before intake, 3.16 ± 0.49 after intake; p = 0.44). All predictor variables were used as dependent variables in a multiple regression model of the incremental change in CFVR after 2 weeks of chocolate intake. Intake of dark (but not white) chocolate, MDA-LDL, triglyceride (TG) and heart rate (HR) significantly influenced the change of CFVR after 2 weeks of intake ( p < 0.01) according to the multiple regression formula: Y = 1.01 X 1 − 0.005 X 2 − 0.003 X 3 − 0.017 X 4 ( Y = change in CFVR after 2 weeks of chocolate intake, X 1 = intake of dark (but not white) chocolate, X 2 = MDA-LDL, X 3 = TG, X 4 = HR). Conclusion Flavonoid-rich dark chocolate intake significantly improved coronary circulation in healthy adults, independent of changes in oxidative stress parameters, blood pressure and lipid profile, whereas non-flavonoid white chocolate had no such effects.

Suppressive Effect of Cocoa Powder on Atherosclerosis in Kurosawa and Kusanagi-hypercholesterolemic Rabbits

We investigated the suppressive effect of cocoa powder (cacao polyphenol content: 7.8%) on atherosclerosis in a spontaneous familial hypercholesterolemic model, Kurosawa and Kusanagi-hypercholesterolemic (KHC) rabbits. Six-month dietary administration of cocoa powder had no effects on body weight, hematology or blood chemistry parameters or a lipid profile in KHC rabbits. Antioxidative activity of low-density lipoprotein (LDL) was observed in the 2nd month and 3rd month of administration. Thiobarbituric acid reactive substances (TBARS), the marker of lipid peroxidation, in plasma were decreased in the cocoa powder treated group from the 2nd month of administration during the study period compared to that in the control group. The area of atherosclerotic lesions in th aorta was significantly smaller in the cocoa powder group (30.87%) than in the control (52.39%). Tissue cholesterol content also tended to decrease. Distensibility of the aortic wall was improved significantly in the cocoa powder treated group due to decreases in fatty streaks and intimal thickening compared to that in the control group. These results suggest that cocoa powder has suppressive effect on development of atherosclerotic lesions. We consider that antioxidative activity of polyphenols rich in cocoa powder may be a key factor for the anti-atherosclerotic effect.