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Antioxidative activity of green tea polyphenol in cholesterol-fed rats.

This study investigated the effects of green tea polyphenol on the serum antioxidative activity and cholesterol levels of cholesterol-fed rats and compared them with those of probucol, an antioxidant hypocholesterolemic agent. To evaluate the antioxidative activity, the susceptibility to oxidative modification of low-density lipoprotein (LDL) isolated from the serum of cholesterol-fed rats was measured, as was the serum antioxidative activity using the spontaneous autoxidation system of brain homogenate. Administration of green tea polyphenol effectively inhibited LDL oxidation and elevated serum antioxidative activity to the same degree as probucol. However, higher amounts of polyphenol than probucol needed to be administered to reduce the total, free, and LDL cholesterol levels. Furthermore, green tea polyphenol increased the levels of high-density lipoprotein (HDL) cholesterol, leading to dose-dependent improvement of the atherogenic index, an effect that was not seen with probucol. Thus, green tea polyphenol may exert an antiatherosclerotic action by virtue of its antioxidant properties and by increasing HDL cholesterol levels.

Animals↗

Inhibitory effects of green tea polyphenols on the production of a virulence factor of the periodontal-disease-causing anaerobic bacterium Porphyromonas gingivalis.

The effect of polyphenolic compounds isolated from green tea (Camellia sinensis) on the production of toxic end metabolites of Porphyromonas gingivalis was investigated. Green tea polyphenols completely inhibited the production of n-butyric acid and propionic acid at a concentration of 1.0-2.0 mg/mL in general anaerobic medium (GAM). (-)-Epigallocatechin gallate (EGCg), which is a major component of tea polyphenols also inhibited the production of phenylacetic acid at 0.5 mg/mL in GAM broth. In the experiment using resting cells of P. gingivalis, phenylacetic acid was produced from l-phenylalanine and phenylpyruvic acid, but this reaction was also inhibited by EGCg, (-)-epicatechin gallate, and (-)-gallocatechin gallate. However, (+)-catechin, (+)-gallocatechin, (-)-epicatechin, and (-)-epigallocatechin did not inhibit those reactions. These results indicate that the inhibitory effect on the production of toxic end metabolites of P. gingivalis can be attributed to the presence of the galloyl moiety, which is ester-linked with the 3-OH of the catechin moiety in the polyphenolic compounds. This study shows that continuous application of tea polyphenols on a daily basis can be considered as a useful and practical method for the prevention of periodontal diseases.

Camellia sinensis↗

Effect of cultivar and processing method on the contents of polyphenols in table olives.

Polyphenols were determined by HPLC in the juice and oil of packed table olives. The phenolic compositions of the two phases were very different, hydroxytyrosol and tyrosol being the main polyphenols in olive juice and tyrosol acetate, hydroxtyrosol acetate, hydroxytyrosol, tyrosol, and lignans (1-acetoxypinoresinol and pinoresinol) in oil. The type of processing had a marked influence on the concentration of polyphenols in olive juice and little on the content in oil. The analyses carried out on 48 samples showed that turning color olives in brine had the highest concentration in polyphenols ( approximately 1200 mg/kg), whereas oxidized olives had the lowest ( approximately 200 mg/kg). Among olive cultivars, Manzanilla had a higher concentration than Hojiblanca and Gordal. The type of olive presentation also influenced the concentration of polyphenols in olives, decreasing in the order plain > pitted > stuffed. The results obtained in this work indicate that table olives can be considered a good source of phenolic antioxidants, although their concentration depends on olive cultivar and processing method.

Chromatography, High Pressure Liquid↗

Use of DSC to detect the heterogeneity of hydrothermal stability in the polyphenol-treated collagen matrix.

The hydrothermal stability of the collagen matrixes treated with plant polyphenols (tannins) depends on not only the strength of the polyphenol-collagen interactions but also the distribution uniformity of polyphenolic molecules within the collagen fibrils. Traditional methods of uniformity tests rely heavily on the expertise of workers and are thus subjective. This paper describes a differential scanning calorimetry (DSC) study of the sheepskin collagen samples treated with hydrolyzable tannins, including two commercial tannins' extracts (chestnut and valonea), two pure ellagitannins (vescalagin and castalagin), and six synthetic gallotannins (di-galloyl-ethylene glycol (DGE), tri-galloyl-glycerol, tetra-galloyl-meso-erythritol, penta-galloyl-adonitol, penta-galloyl-glucose, and hexa-galloyl-ducitol). The collagen sample without polyphenol treatment and the sample treated with DGE showed a single sharp peak in their DSC thermogram with a full peak width at half height (fwhh) of 3-4 degrees C. The samples treated with other tannins all showed multiple peak DSC profiles with the fwhh of each peak at about 3-4 degrees C. These multiple peak profiles imply that in these polyphenol-treated samples, there is a distribution of collagen molecules having different hydrothermal stability. The results have demonstrated that DSC offers an objective method to detect the stability heterogeneity of collagen matrixes in the solid state, providing a useful tool for the leather industry to evaluate the uniformity of leather tanning.

Animals↗

Polyphenolic compounds, antioxidant capacity, and quinone reductase activity of an aqueous extract of Ardisia compressa in comparison to mate (Ilex paraguariensis) and green (Camellia sinensis) teas.

Aqueous extracts of the leaves of Ardisia compressa (AC) have been used in folk medicine to treat various liver disorders including liver cancer. The objective of this study was to partially characterize and determine the total polyphenol content, antioxidant capacity, and quinone reductase activity of A. compressa tea in comparison to mate (Ilex paraguariensis, MT) and green (Camellia sinensis,GT) teas. Total polyphenol content, antioxidant capacity, and phase II enzyme induction capacity were measured by the modified Folin-Ciocalteu, ORAC, and quinone reductase (QR) assays, respectively. The major polyphenols in AC were not catechins. HPLC retention times and standard spikes of AC indicated the presence of gallic acid, epicatechin gallate, ardisin and kaempferol. Using catechin as standard, the total polyphenol value of AC (36.8 +/- 1.1 mg/mg DL) was significantly lower than GT (137.2 +/- 5.8 mg equivalent of (+)-catechin/mg dried leaves, DL) and MT (82.1 +/- 3.8 mg/mg DL) (P < 0.001). Antioxidant capacity (AC, 333; GT, 1346; MT, 1239 mmol Trolox equivalents/g DL) correlated with total polyphenol values (r(2) = 0.86, P < 0.01). AC (4.5-12.5 microg/mL) induced QR enzyme, in Hepa1c1c7 cells, up to 15%. MT and GT showed no induction at the concentrations tested (0.5-10.5 and 0.5-12.5 mg/mL, respectively). These results suggest that AC has a different mechanism of protection against cytotoxicity that is not related to its antioxidant capacity. Further studies are needed to determine such mechanisms and to explore its potential as a chemopreventive or therapeutic agent.

Antioxidants↗

Polyphenolic profiles of Basque cider apple cultivars and their technological properties.

The polyphenolic compositions of 31 Basque cider apple cultivars were determined in pulp, peel, and juice by high-performance liquid chromatography with diode array detection analysis of crude extracts and after thiolysis. Total polyphenols are distributed in a wide concentration range depending on the cultivar. Procyanidins are the class of polyphenols that present major concentrations in apple. Their average degrees of polymerization range from 4 to 8 depending on the cultivar. Apple cultivars were technologically classified into bitter and nonbitter categories using different classification systems obtained by applying several pattern recognition techniques, such as principal component analysis, K-nearest neighbors, soft independent modeling of class analogy, partial least-squares, and multilayer feed-forward-artificial neural networks, to apple pulp, peel, or juice data (individual polyphenol concentrations, total procyanidin content, and the average degree of polymerization of procyanidins). Bitter apple cultivars present higher contents of flavan-3-ols and/or dihydrochalcones than nonbitter cultivars. Detailed knowledge of the polyphenolic profile of each apple cultivar affords information about their susceptibility to oxidation, their sensory properties (bitterness, astringency), and their possible influence on the characteristics and quality of the final product (juice, cider) when apples are processed.

Beverages↗

Polyphenol-rich phloem enhances the resistance of total serum lipids to oxidation in men.

In humans, polyphenol supplementation studies have resulted in inconsistent findings in lipid peroxidation. Our aim was to investigate the effects of a 4-week consumption of polyphenol-rich phloem on serum lipids and lipid peroxidation in the hydrophilic fraction of serum and on isolated lipoproteins. We conducted a randomized double-blind supplementation study consisting of 75 nonsmoking hypercholesterolemic men. Participants consumed 70 g daily of either rye bread (placebo) or phloem-fortified rye bread containing 31 mg (low polyphenol, LP) or 62 mg (high polyphenol, HP) of catechins. The ex vivo susceptibility of total serum lipids and VLDL and LDL to oxidation after copper induction was measured as a lag time to the maximal oxidation rate at the baseline and after the supplementation. In the HP group, an increase in the oxidation resistance of total serum lipids was observed (11.4%), while no effect was seen in the LP group (-0.8%) or in the placebo group (-1.0%) (p = 0.007). No differences were observed in the oxidation resistance of VLDL and LDL between the study groups. The phloem also increased in vitro oxidation resistance of serum lipids and radical scavenging activity (DPPH.) in a dose-dependent manner. Our results suggest that polyphenols may inhibit lipid peroxidation in the hydrophilic fraction of serum.

Adult↗

Quantitation of polyphenols in different apple varieties.

Forty-one apple samples, representing eight of the most widely cultivated varieties in western Europe, were collected in Trentino, Italy. Samples were extracted from fresh fruit with a mixture of acetone/water to achieve a good extraction of polyphenols, including proanthocyanidin oligomers which were analyzed by normal-phase HPLC. Up to 20 compounds including catechin, epicatechin, B2 procyanidin, hydroxycinnamates, flavonols, anthocyanins, and dihydrochalcones were analyzed by reversed-phase HPLC and LC-MS. Total polyphenol content was independently measured with an optimized Folin-Ciocalteu assay. The mean content of total polyphenols lay between 66.2 and 211.9 mg/100 g of FW depending on the variety. With chromatographic analysis, it was possible to explain the whole amount of total polyphenols measured by the FC assay. Flavanols (catechin and proanthocyanidins) are the major class of apple polyphenols (71-90%), followed by hydroxycinnamates (4-18%), flavonols (1-11%), dihydrochalcones (2-6%), and in red apples anthocyanins (1-3%).

Chromatography, High Pressure Liquid↗

Effect of mash maceration on the polyphenolic content and visual quality attributes of cloudy apple juice.

The effects of enzymatic mash treatments on yield, turbidity, color, and polyphenolic content of cloudy apple juice were studied. Using HPLC-ESI-MS, cryptochlorogenic acid was identified in cv. Brettacher cloudy apple juice for the first time. Commercial pectolytic enzyme preparations with different levels of secondary protease activity were tested under both oxidative and nonoxidative conditions. Without the addition of ascorbic acid, oxidation substantially decreased chlorogenic acid, epicatechin, and procyanidin B2 contents due to enzymatic browning. The content of chlorogenic acid as the major polyphenolic compound was also influenced by the composition of pectolytic enzyme preparations because the presence of secondary protease activity resulted in a rise of chlorogenic acid. The latter effect was probably due to the inhibited protein-polyphenol interactions, which prevented binding of polyphenolic compounds to the matrix, thus increasing their antioxidative potential. The results obtained clearly demonstrate the advantage of the nonoxidative mash maceration for the production of cloud-stable apple juice with a high polyphenolic content, particularly in a premature processing campaign.

Beverages↗

Ethanol and polyphenolic free wine matrix stimulate the differentiation of human intestinal Caco-2 cells. Influence of their association with a procyanidin-rich grape seed extract.

The effect of daily contact with ethanol on Caco-2 cell differentiation was investigated. Pure ethanol (1%) and a polyphenolic free wine matrix (polyphenol-free wine containing 1% ethanol) associated or not with a procyanidin-rich grape seed extract (GSE) were added to Caco-2 cells from confluency for 2 h a day after successive incubation in salivary, gastric, and pancreatic media. Treatment with 1% ethanol did not appear to be cytotoxic to cells, but it also stimulated Caco-2 cell differentiation, particularly in the first days following confluency, and this effect was more marked when associated with polyphenolic free wine matrix constituents. This activation resulted in an increase in microvillar density, organization, and elongation (+70%) and was associated with strong stimulation of sucrase-isomaltase (+780%) and a concomitant regular increase in cell protein content (+50-88%). While the presence of GSE in alcoholic solutions did not modify the morphological pattern observed in cells subjected to ethanol and polyphenolic free wine matrix alone, it had a clear reducing effect on their microvillus elongation (-30%). However, these stimulating effects of ethanol on morphological differentiation were attenuated from day 10 postconfluency, which could suggest cell cytoprotection against ethanol. These are the first results in support of the notion that moderate concentration of ethanol may stimulate the differentiation of Caco-2 cells, particularly when integrated with a polyphenolic free wine matrix.

Biflavonoids↗

Açai (Euterpe oleracea Mart.) polyphenolics in their glycoside and aglycone forms induce apoptosis of HL-60 leukemia cells.

The effects of açai polyphenolics on the antiproliferation and induction of apoptosis in HL-60 human leukemia cells were investigated. Interactions between anthocyanins and non-anthocyanin-polyphenolics in both their glycosidic and their aglycone forms were also investigated to determine additive or nonadditive responses. Polyphenolic fractions at 0.17-10.7 microM were found to reduce cell proliferation from 56 to 86% likely due to caspase-3 activation (apoptosis). Anthocyanin and polyphenolic fractions were nonadditive in their contribution to the cell antiproliferation activity. At equimolar concentrations, the glycosidic forms of phenolic acids and flavonoids induced a higher magnitude of change in cell parameters (proliferation and apoptosis) than their respective aglycone forms, while the opposite trend was observed for anthocyanin aglycones. This study demonstrated that açai offers a rich source of bioactive polyphenolics and confirmed the importance of investigating whole food systems when evaluating the potential health benefits of individual phytochemical compounds.

Anthocyanins↗

Modulation of key elements of the Wnt pathway by apple polyphenols.

Glycogen synthase kinase-3beta (GSK3beta) is one of the key elements of the Wnt pathway involved in the regulation of beta-catenin homeostasis. The inhibition of GSK3beta kinase activity might lead to the onset of beta-catenin/TCF/LEF-mediated gene transcription, representing a potentially mitogenic stimulus. Apple polyphenols have been shown to mediate several biological effects that might be of interest with respect to chemoprevention. The objective of the study was to elucidate whether apple polyphenols also modulate key elements of the Wnt pathway, an effect that might limit the usefulness of these compounds for the prevention of carcinogenesis. A polyphenol-rich apple juice extract (AE02) was found to effectively inhibit the kinase activity of GSK3beta, immunoprecipitated from HT29 cells. Treatment of HT29 cells with AE02 for 24 h resulted in a concentration-dependent decrease of the cellular GSK3beta kinase activity. The inhibition of the kinase activity in HT29 cells was observed at polyphenol concentrations corresponding to the concentration of the constituents in the original apple juice. The apple characteristic dihydrochalcone glycoside phloridzin was found to be inactive up to 500 muM. The free aglycon phloretin as well as the flavonol quercetin effectively inhibited isolated GSK3beta, but did not affect the respective kinase activity within HT29 cells. In accordance with the inhibition of GSK3beta kinase activity by AE02, treatment of HT29 cells resulted in a significant decrease of phosphorylated beta-catenin. However, the total intracellular beta-catenin level was also found to be diminished, indicating that the interference of the apple constituents with GSK3beta was not associated with a stabilization of beta-catenin in HT29 cells. In line with these results, TCF/LEF-mediated gene transcription remained unaffected by treatment with AE02 as shown in a reporter gene approach. We can assume from the results that at consumer-relevant concentrations apple polyphenols do not mediate growth-stimulating effects in HT29 cells via the Wnt pathway.

Blotting, Western↗

Treatment of mcf-7 breast cancer cells with a red grape wine polyphenol fraction results in disruption of calcium homeostasis and cell cycle arrest causing selective cytotoxicity.

Food components influence the physiology by modulating gene expression and biochemical pathways within the human body. The disease-preventive roles of several fruit and vegetable components have been related to such properties. Polyphenolic components such as flavonoids are strong antioxidants and induce the expression of several xenobiotic-detoxifying enzymes. The mechanism of selective cytotoxicity induced by red grape wine polyphenols against MCF-7 breast cancer cells was investigated in relation to their interference with calcium homeostasis. MCF-7 cells showed an increase in cytosolic calcium levels within 10 min of treatment with the polyphenols. Immunohistochemical localization of calmodulin with secondary gold-labeled antibodies showed similar levels of gold labeling in both MCF-7 cells and the spontaneously immortalized, normal MCF-10A cell line. MCF-7 cells treated with the red wine polyphenol fraction (RWPF) showed swelling of endoplasmic reticulum, dissolution of the nucleus, and loss of plasma membrane integrity as well as reduced mitochondrial membrane potential. These cells were arrested at the G2/M interphase. By contrast, MCF-10A cells did not show such changes after RWPF treatment. The results suggest that polyphenol-induced calcium release may disrupt mitochondrial function and cause membrane damage, resulting in selective cytotoxicity toward MCF-7 cells. This property could further be developed toward breast cancer prevention strategies either independently or in conjunction with conventional prevention therapies where a positive drug-nutrient interaction can be demonstrated.

Breast Neoplasms↗

Enhancement of polyphenol recovery from rosemary (Rosmarinus officinalis) and sage (Salvia officinalis) by enzyme-assisted ensiling (ENLAC).

The efficacy of enzyme-assisted ensiling (ENLAC) in the recovery of polyphenols from rosemary and sage was tested. Fresh rosemary and sage were chopped and ensiled in 0.5-L anaerobic jars. Treatments comprised control (no additives), 0.5% glucose and lactic acid bacteria, and 1% cellulase plus 1% hemicellulase plus pectinase. Following storage at room temperature for 45 days (experiment 1) and 26 days (experiment 2), polyphenols were extracted from the silages in ethanol either by direct blending or by cold extraction. The enzyme treatment resulted in silages with the lowest pH values, lowest fiber content, highest water-soluble sugar content, and highest polyphenol recovery; this treatment resulted in increased polyphenol recovery from rosemary and sage, by 100 and 20%, respectively. Comparison between direct blending and cold extraction revealed similar efficiency of polyphenol recovery.

Enzymes↗

Comparison of natural polyphenol antioxidants from extra virgin olive oil with synthetic antioxidants in tuna lipids during thermal oxidation.

Polyphenols extracted from extra virgin olive oil (EVOO) were tested for their ability to inhibit lipid oxidation of canned tuna. Hydroperoxide formation during oxidation was monitored by measurement of peroxide value and decomposition of hydroperoxides by static headspace gas chromatographic analysis of volatiles. In tuna oxidized at 40 and 100 degrees C, 400 ppm of the EVOO polyphenols was an effective antioxidant as compared with 100 ppm of a 1:1 mixture of the synthetic antioxidants butylated hydroxytoluene and butylated hydroxyanisole. However, at concentrations <100 ppm, the EVOO phenolic compounds promoted hydroperoxide formation and decomposition. The EVOO polyphenols were effective antioxidants when added to heated tuna muscle in the presence of either brine or refined olive oil. The oxidation rate in tuna muscle packed in brine was higher than that of tuna packed in refined olive oil. The EVOO polyphenols had higher antioxidant activity in the brine samples than in the refined olive oil. The higher antioxidant activity of EVOO polyphenols in tuna packed in brine may be explained by their greater affinity toward the more polar interface between water and the fish oil system.

Animals↗

Red wine polyphenols increase calcium in bovine aortic endothelial cells: a basis to elucidate signalling pathways leading to nitric oxide production.

1. The present study investigates the mechanisms by which polyphenolic compounds from red wine elicit Ca(2+) mobilization in bovine aortic endothelial cells (BAECs). Two polyphenol-containing red wine extracts, red wine polyphenolic compounds (RWPC) and Provinols, and delphinidin, an anthocyanin were used. 2. RWPC stimulated a Ca(2+)-dependent release of nitric oxide (NO) from BAECs accounting for the relaxation of endothelium-denuded rat aortic rings as shown by cascade bioassay. 3. RWPC, Provinols and delphinidin increased cytosolic free calcium ([Ca(2+)](i)), by releasing Ca(2+) from intracellular stores and by increasing Ca(2+) entry. 4. The RWPC-induced increase in [Ca(2+)](i) was decreased by exposure to ryanodine (30 microM), whereas Provinols and delphinidin-induced increases in [Ca(2+)](i) were decreased by bradykinin (0.1 microM) and thapsigargin (1 microM) pre-treatment. 5. RWPC, Provinols and delphinidin-induced increases in [Ca(2+)](i) were sensitive to inhibitors of phospholipase C (neomycin, 3 mM; U73122, 3 microM) and tyrosine kinase (herbimycin A, 1 microM). 6. RWPC, Provinols and delphinidin induced herbimycin A (1 microM)-sensitive tyrosine phosphorylation of several intracellular proteins. 7. Provinols released Ca(2+) via both a cholera (CTX) and pertussis toxins (PTX)-sensitive pathway, whereas delphinidin released Ca(2+) only via a PTX-sensitive mechanism. 8. Our data contribute in defining the mechanisms of endothelial NO production caused by wine polyphenols including the increase in [Ca(2+)](i) and the activation of tyrosine kinases. Furthermore, RWPC, Provinols and delphinidin display differences in the process leading to [Ca(2+)](i) increases in endothelial cells illustrating multiple cellular targets of natural dietary polyphenolic compounds.

Animals↗

Inhibition of cardiac voltage-gated sodium channels by grape polyphenols.

BACKGROUND AND PURPOSE: The cardiovascular benefits of red wine consumption are often attributed to the antioxidant effects of its polyphenolic constituents, including quercetin, catechin and resveratrol. Inhibition of cardiac voltage-gated sodium channels (VGSCs) is antiarrhythmic and cardioprotective. As polyphenols may also modulate ion channels, and possess structural similarities to several antiarrhythmic VGSC inhibitors, we hypothesised that VGSC inhibition may contribute to cardioprotection by these polyphenols. EXPERIMENTAL APPROACH: The whole-cell voltage-clamp technique was used to record peak and late VGSC currents (INa) from recombinant human heart NaV1.5 channels expressed in tsA201 cells. Right ventricular myocytes from rat heart were isolated and single myocytes were field-stimulated. Either calcium transients or contractility were measured using the calcium-sensitive dye Calcium-Green 1AM or video edge detection, respectively. KEY RESULTS: The red grape polyphenols quercetin, catechin and resveratrol blocked peak INa with IC50s of 19.4 microM, 76.8 microM and 77.3 microM, respectively. In contrast to lidocaine, resveratrol did not exhibit any frequency-dependence of peak INa block. Late INa induced by the VGSC long QT mutant R1623Q was reduced by resveratrol and quercetin. Resveratrol and quercetin also blocked late INa induced by the toxin, ATX II, with IC50s of 26.1 microM and 24.9 microM, respectively. In field-stimulated myocytes, ATXII-induced increases in diastolic calcium were prevented and reversed by resveratrol. ATXII-induced contractile dysfunction was delayed and reduced by resveratrol. CONCLUSIONS AND IMPLICATIONS: Our results indicate that several red grape polyphenols inhibit cardiac VGSCs and that this effect may contribute to the documented cardioprotective efficacy of red grape products.

Animals↗

Polyphenol-enriched oolong tea increases fecal lipid excretion.

OBJECTIVE: To assess possibility of polyphenol-enriched oolong tea to reduce dietary lipid absorption in humans. DESIGN: Twelve healthy adult subjects, three males and nine females, aged (mean+/-s.d.) 22.0+/-1.8 years, respectively, were randomly divided into two groups. The participants were followed a double-blind placebo-controlled crossover design, including 7-day washout periods and 10-day treatment periods. During the treatment periods, subjects were given about 38 g of lipids from potato chips (19 g each within 30 min after lunch and dinner) and total 750 ml beverages (placebo- or polyphenol-enriched oolong tea) at three meals. Blood samples were collected for biochemical examination at days 8, 18, 25 and 35 of the study period. On the last 3 days of each treatment period, feces were collected to measure the excretion of lipids. RESULTS: Lipid excretion into feces was significantly higher in the polyphenol-enriched oolong tea period (19.3+/-12.9 g/3 day) than in the placebo period (9.4+/-7.3 g/3 day) (P < 0.01). Cholesterol excretion tended to increase in polyphenol-enriched oolong tea period (1.8+/-1.2 g/3 day) compared with that of placebo (1.2+/-0.6 g/3 day) (P = 0.056). CONCLUSIONS: The results of this study indicated that polyphenol-enriched oolong tea could increase lipid excretion into feces when subjects took high-lipid diet.

Adult↗