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PubMed2006

Improvement of leukocyte functions in prematurely aging mice after five weeks of diet supplementation with polyphenol-rich cereals.

OBJECTIVE: We investigated the beneficial effects of diet supplementation with two types of cereals naturally rich in polyphenolic compounds on several functions of peritoneal leukocytes from prematurely aging mice (PAM). METHODS: Two-hundred sixty healthy mice, 8 wk of age, were recruited and their behavioral responses were tested in a simple T-maze to identify PAM. Then the mice were fed a diet supplemented with 20% (wt/wt) of two different cereal fractions, named B (wheat germ) and C (buckwheat flour), rich in polyphenols (gallic acid, catechin, p-hydroxybenzoic acid, vanillic acid, p-coumaric acid, sinapic acid, ferulic acid, quercetin, and rutin), or a standard diet (controls) for 5 wk. Several parameters of innate (adherence to tissues, chemotaxis, phagocytosis, microbicidal capacity, and natural killer activity) and acquired immune (lymphoproliferation and interleukin-2 release) responses were measured. RESULTS: The PAM control group showed worse immune functions (P < 0.001 to 0.05) compared with the non-PAM control group. The PAM group that received cereal B showed increases in phagocytosis (P < 0.01), microbicidal activity (P < 0.001 to 0.01), natural killer activity (P < 0.001) and lymphoproliferation in response to lipopolysaccharide (P < 0.01) and interleukin-2 release (P < 0.001), and the PAM group that received cereal C showed a similar pattern, with increases in macrophage chemotaxis (P < 0.01), phagocytosis (P < 0.01), microbicidal activity (P < 0.001 to 0.01), natural killer activity (P < 0.01), lymphoproliferative response to concanavalin A and lipopolysaccharide (P < 0.001), and interleukin-2 release (P < 0.001). CONCLUSIONS: Dietary supplementation with polyphenol-rich cereals appears to have a protective effect on immune cell functions in mice with premature senescence. Thus, regular intake of these compounds could delay normal aging and improve quality of life.

Aging↗
PubMed2005

Potent suppressive effect of green tea polyphenols on tobacco-induced mutagenicity.

Antimutagenic activity of green tea (Camellia sinensis) was studied using Salmonella typhimurium strains (TA 102) (Ames test). Aqueous tobacco extract was found to be mutagenic to S. typhimurium TA 102 at concentration of 50 mg/plate. Green tea polyphenols was found to inhibit the mutagenicity of tobacco in a concentration-dependent manner. Concentrations needed for 50% inhibition of mutagen-induced revertant formation was found to be 5 mg/plate. Green tea polyphenols was also found to inhibit the urinary mutagenicity in rats induced by tobacco extract. Moreover green tea polyphenols were found to inhibit in vitro nitrosation reaction produced by reaction sodium nitrite and methyl urea and further inhibition of mutagenicity indicating that green tea has dual action to bring out a reduction in the mutagenic and carcinogenic potential of tobacco.

Animals↗
PubMed2004

Leishmanicidal activity of polyphenolic-rich extract from husk fiber of Cocos nucifera Linn. (Palmae).

The available therapy for leishmaniasis, which affects 2 million people per annum, still causes serious side effects. The polyphenolic-rich extract from the husk fiber of Cocos nucifera Linn. (Palmae) presents antibacterial and antiviral activities, also inhibiting lymphocyte proliferation, as shown by our group in previous works. In the present study, the in vitro leishmanicidal effects of C. nucifera on Leishmania amazonensis were evaluated. The minimal inhibitory concentration of the polyphenolic-rich extract from C. nucifera to completely abrogate parasite growth was 10 microg/ml. Pretreatment of peritoneal mouse macrophages with 10 microg/ml of C. nucifera polyphenolic-rich extract reduced approximately 44% the association index between these macrophages and L. amazonensis promastigotes, with a concomitant increase of 182% in nitric oxide production by the infected macrophage in comparison to nontreated macrophages. These results provide new perspectives on drug development against leishmaniasis, since the extract of C. nucifera at 10 microg/ml is a strikingly potent leishmanicidal substance which inhibited the growth of both promastigote and amastigote developmental stages of L. amazonensis after 60 min, presenting no in vivo allergenic reactions or in vitro cytotoxic effects in mammalian systems.

Animals↗
PubMed1999

Cellular thiols status and cell death in the effect of green tea polyphenols in Ehrlich ascites tumor cells.

Epidemiological studies suggest that the consumption of green tea may help prevent cancers in humans, and also breast and prostate cancers in animal models are reduced by green tea, and several mechanisms have been proposed for these effects. In this study the relationship between cellular sulfhydryl (SH) groups and the cytotoxicity of green tea polyphenols in Ehrlich ascites tumor cells was examined. It was found that in the presence of green tea extract (GTE) (100 microg/ml) and one of its polyphenolic components, epigallocatechin (EGC; 100 microM), both cellular non-protein (GSH) and protein-sulfhydryl (PSH) levels were significantly decreased and this was associated with a decrease in cell viability. Replenishing the thiol levels by using N-acetylcysteine (NAC) caused a recovery in cell viability, but this recovery was dependent on the time of thiol replenishment in the presence of EGC (initial 15 min). These results identify SH groups as a novel target of green tea polyphenols cytotoxicity in tumor cells, and a regulatory role for green tea in terms of reducing sulfhydryls in tumor inhibition.

Acetylcysteine↗
PubMed2003

Plant polyphenols inhibit VacA, a toxin secreted by the gastric pathogen Helicobacter pylori.

VacA is a major virulence factor of the widespread stomach-dwelling bacterium Helicobacter pylori. It causes cell vacuolation and tissue damage by forming anion-selective, urea-permeable channels in plasma and endosomal membranes. We report that several flavone derivatives and other polyphenols present in vegetables and plants inhibit ion and urea conduction and cell vacuolation by VacA. Red wine and green tea, which contain many of the compounds in question, also potently inhibit the toxin. These observations suggest that polyphenols or polyphenol derivatives may be useful in the prevention or cure of H. pylori-associated gastric diseases.

Anti-Bacterial Agents↗
PubMed2003

Separation of polyphenolic compounds extracted from plant matrices using capillary electrophoresis.

Phenolic compounds constitute a large group of secondary plant products whose chemical structure may range from quite simple compounds to highly polymerized substances. The polyphenols content have been investigated in the alcoholic extract of the fruits of three different plants: sweet gale, sea buckthorn, hiprose. The trans-resveratrol content we have studied in roots, stems, leaves and flowers of Japanese knotweed grown in Estonia. Plant material was pre-treated in two different ways: by infusing with methanol and by supercritical fluid extraction with carbon dioxide modified with different alcohols. The relationship between variables (pressure, temperature, modifier amount) and yields are examined. The capillary zone electrophoresis methods were developed for the separation of polyphenolic anti-oxidative compounds. Using both water based borate buffer and acetonitrile based non-aqueous media it was possible to get reliable separation of several polyphenolic compounds. Based on that there has been identified such as flavone, trans-resveratrol, catechin, chlorogenic acid, quercetin and myricetin in plant extracts. Changes in the relative concentrations of trans-resveratrol in different parts of the knotweed have been established.

Electrophoresis, Capillary↗
PubMed2001

Anticlastogenic effects of black tea (World blend) and its two active polyphenols theaflavins and thearubigins in vivo in Swiss albino mice.

This study investigated the inhibition of cyclophosphamide (CP) and dimethylbenz(a)anthracene (DMBA) induced genetic damage by black tea (World blend) and its two active polyphenols theaflavins (TF) and thearubigins (TR) in Swiss albino mice as measured by chromosome aberrations (CA) and sister chromatid exchanges (SCE). Three different concentrations (5, 10 and 20%) of tea and a single dose of TF and TR were tested for their anticlastogenic effects against DMBA (50 mg/kg body weight) and CP (20 mg/kg for CA and 10 mg/kg for SCE). A significant decrease in CA was observed in all the three concentrations of tea extract plus DMBA treated groups when compared with the respective DMBA treated group alone. Similarly a significant decrease in CA was observed in all the three concentrations of tea extracts plus CP treated series when compared with the group treated with CP alone. In SCE assay, a significant decrease in SCE was observed in 5, 10 and 20% black tea extract plus CP and 10 and 20% tea extracts plus DMBA treated groups when compared with the CP or DMBA treated group alone. In the single dose of TF and TR treated groups a significant decrease in both CA and SCE was observed in both the TF and TR plus both the carcinogen treated groups when compared with their positive controls. The protective effects of black tea extracts were more significant than that of its two polyphenols. This study indicates that both black tea and its active polyphenols TF and TR have significant anticlastogenic effects in bone marrow cells of mice.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed2000

Changes in grape seed polyphenols during fruit ripening.

The quantity and characterization of extracted flavan-3-ol monomers and procyanidins was determined in seeds from Vitis vinifera cv. Cabernet Sauvignon berries, over the course of ripening and at different levels of vine water status. The per berry extractive yield of all polyphenols decreased with maturity, and followed second-order kinetics. The flavan-3-ol monomers decreased most rapidly, followed by the procyanidin extension units and finally, the terminal units. The relative proportion of procyanidin extension units did not vary with maturity. During fruit ripening, the mean degree of polymerization of extracted procyanidins is unchanged when analyzed intact by HPLC, but decreases by thiolytic degradation. The proportion of extracted procyanidins resistant to acid catalyzed thiolysis increased with maturity. Changes in vine water status affected polyphenol amounts, indicating that cultural practices can be used to influence composition. Oxidation of the seed polyphenols during fruit ripening, could explain these observations.

Flavonoids↗
PubMed2001

Determination of quantity and quality of polyphenol antioxidants in foods and beverages.

The methods described in this article are quick, simple, and inexpensive to perform. The Folin quantitation method can determine both free and total polyphenol antioxidants in foods and beverages as described, as well as botanical extracts. This assay may also be used to estimate the daily per capita consumption of polyphenols in foods. The dose-response in vitro lower density lipoprotein antioxidant activity measurement (IC50) can be employed to compare antioxidants as pure compounds, or in mixtures after quantitating the polyphenols. The ex vivo lipoprotein-binding antioxidant activity can be measured simply and rapidly to determine possible in vivo binding of pure compounds or extracts from foods. Supplementation and epidemiology studies can utilize the rapid and inexpensive affinity column isolation method of lower density lipoproteins for the determination of lipoprotein oxidative susceptibility.

Antioxidants↗
PubMed1996

Inhibition of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) mutagenicity by black and green tea extracts and polyphenols.

Solutions of lyophilized preparations of standard black and green tea extracts were made and tested over a range of six concentrations as inhibitors of the mutagenicity caused by the fool mutagen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in the Salmonella typhimurium TA98 assay containing S9 fraction from rats induced with alpha-naphthoflavone and phenobarbital. Extracts of both black and green tea were equally good inhibitors of mutagenicity. Purified polyphenols were prepared from tea extracts by solvent extraction. The polyphenols of black tea were more potent inhibitors of mutagenicity than the polyphenols of green tea. These findings suggest that black tea may have similar health-promoting properties to those reported previously for green tea.

Animals↗
PubMed2001

Anticlastogenic activity of cacao: inhibitory effect of cacao liquor polyphenols against mitomycin C-induced DNA damage.

Oxidative DNA damage has been implicated as a factor playing a role in mutagenesis and carcinogenesis. We investigated the anticlastogenic activity of cacao: the inhibitory effect of cacao liquor polyphenols on DNA strand cleavage induced by mitomycin C (MMC) in vitro and the anticlastogenic effect of cacao liquor extract against formation of micronuclei induced by MMC in bone marrow cells and peripheral blood cells of mice. In the DNA strand cleavage test, cacao liquor polyphenols inhibited cleavage of RFI DNA. In the micronuclei test, the frequency of occurrence of micronucleated cells among bone marrow cells and peripheral blood cells were reduced significantly when cacao liquor extract was administered orally to mice 6 h before intraperitoneal injection of MMC. These findings suggest that cacao liquor polyphenols are effective in preventing DNA damage, and one of the mechanisms of action might involve scavenging of active oxygen radicals generated in reactions initiated by MMC.

Animals↗
PubMed2002

Higher radical scavenging activities of polyphenolic antioxidants can be ascribed to chemical reactions following their oxidation.

Radical scavenging activities of 34 natural antioxidants were investigated from an electrochemical viewpoint. While the correlation of the oxidation potentials with their DPPH radical scavenging activities (represented by EC(50)) was not high (the correlation coefficient, r=0.73), the number of electrons n required for oxidation of an antioxidant, being obtained by continuous flow-column electrolysis with a slower flow rate (0.05 ml min(-1)), did show a good correlation with EC(50) (1/EC(50)=1.67n+0.50 with r=0.94). The n values of most polyphenols were increased with a decrease in the flow rate, while those of nonpolyphenols were invariant. This suggests that a slower subsequent chemical reaction(s) should be involved in the oxidation of polyphenols, whose higher radical scavenging activities seem to be ascribed to the chemical reactions. In this study, we have proposed a possible mechanism for the oxidation of polyphenols, in which the oxidizable -OH moieties are reproduced through an oxidative dimerization (or more highly polymerization).

Antioxidants↗
PubMed1998

Polyphenol-mediated inhibition of AP-1 transactivating activity may slow cancer growth by impeding angiogenesis and tumor invasiveness.

Certain polyphenols, capable of activating transcription via antioxidant response elements, potently induce Fra-1/Fra-2, thereby antagonizing the transactivating activity of AP-1 complexes. Since AP-1 is crucial to the induction of matrix proteases required for angiogenesis and tumor invasion, it is reasonable to suspect that adequate intakes of appropriate polyphenols would have utility in cancer therapy. Indeed, green tea polyphenols have reduced tumor growth rate and/or metastatic capacity in several animal studies. Additional measures for down-regulating AP-1 activity may include fish oil, heparin, and the drug LY290181. Down-regulation of AP-1 activity as a general therapeutic strategy merits further consideration.

Animals↗
PubMed1997

Application of a new high-performance liquid chromatographic method for measuring selected polyphenols in human plasma.

We developed a method to measure plasma levels of selected polyphenols before and after ingestion of green tea. Blood samples were obtained from four healthy women before and 30 and 50 min after the ingestion of 300 ml of green tea infusion. A 1-ml volume of plasma was hydrolysed with 0.5 M HCl-methanol (1:1, v/v) for 30 min at room temperature, extracted with ethyl acetate and separated by reversed-phase chromatography. Polyphenols were identified on the basis of their retention times and by spectrum analysis. Green tea caffeine has the same retention times as caffeic acid. Consumption of green tea produces a notable increase in the plasma levels of caffeine plus caffeic acid and the appearance of measurable levels of epigallocatechingallate. In conclusion, the method was found to have the requisite features of specificity and sensitivity for monitoring plasma levels of selected tea polyphenols.

Chromatography, High Pressure Liquid↗
PubMed2003

Novel liquid chromatography-electrospray ionization mass spectrometry method for the quantification in human urine of microbial aromatic acid metabolites derived from dietary polyphenols.

An HPLC-ESI-MS-MS method was developed to quantify in human urine fourteen aromatic acids known as metabolites of dietary polyphenols. These metabolites were determined simultaneously in a single 20-min chromatographic analysis with multiple reaction monitoring detection. The inter- and intra-day precisions, calculated from quality control samples were 8.8 and 5.3%, respectively, and the mean accuracy was 2.3%. The method was tested on urine samples collected from one healthy volunteer who consumed a polyphenol-rich diet for 3 days. Increased levels of several aromatic acid metabolites were observed, demonstrating that the method can be used to detect changes in the excretion of microbial metabolites induced by the consumption of polyphenol-containing foods in humans.

Acids↗
PubMed2000

Preparative separation of polyphenols from tea by high-speed countercurrent chromatography.

High-speed countercurrent chromatography (HSCCC) was applied to the separation of polyphenols from tea leaves (Camellia sinensis L.). The capability of HSCCC to isolate pure tea polyphenols from complex mixtures on a preparative scale was demonstrated for catechins, flavonol glycosides, proanthocyanidins, and strictinin from green and black tea. The purity and identity of isolated compounds was confirmed by (1)H NMR and HPLC-ESI-MS/MS. Gram quantities of polyphenols from tea can be isolated with the procedure described.

Chromatography, High Pressure Liquid↗
PubMed2000

Induction of apoptosis by the oolong tea polyphenol theasinensin A through cytochrome c release and activation of caspase-9 and caspase-3 in human U937 cells.

This study examined the growth inhibitory effects of theasinensin A (from oolong tea) and black tea polyphenols, including theaflavin (TF-1), a mixture (TF-2) of theaflavin-3-gallate (TF-2a) and theaflavin-3'-gallate (TF-2b), and theaflavin-3,3'-digallate (TF-3) in human cancer cells. Theasinensin A, TF-1, and TF-2 displayed strong growth inhibitory effects against human histolytic lymphoma U937, with estimated IC50 values of 12 microM, but were less effective against human acute T cell leukemia Jurkat, whereas TF-3 and (-)-epigallocatechin-3-gallate (EGCG) had lower activities. The molecular mechanisms of tea polyphenol-induced apoptosis as determined by annexin V apoptosis assay, DNA fragmentation, and caspase activation were further investigated. Loss of membrane potential and reactive oxygen species (ROS) generation were also detected by flow cytometry. Treatment with tea polyphenols caused rapid induction of caspase-3, but not caspase-1, activity and stimulated proteolytic cleavage of poly(ADP-ribose) polymerase (PARP). Pretreatment with a potent caspase-3 inhibitor, Z-Asp-Glu-Val-Asp-fluoromethyl ketone, inhibited theasinensin A induced DNA fragmentation. Furthermore, it was found that theasinensin A induced loss of mitochondrial transmembrane potential, elevation of ROS production, release of mitochondrial cytochrome c into the cytosol, and subsequent induction of caspase-9 activity. These results indicate that theasinensin A allows caspase-activated deoxyribonuclease to enter the nucleus and degrade chromosomal DNA and induces DFF-45 (DNA fragmentation factor) degradation. The results suggest that induction of apoptosis by theasinensin A may provide a pivotal mechanism for their cancer chemopreventive function.

Apoptosis↗
PubMed2001

Determination of polyphenolic profiles of Basque cider apple varieties using accelerated solvent extraction.

Polyphenols in the peel and pulp of 15 Basque cider apple varieties were determined by accelerated solvent extraction followed by reversed phase high-performance liquid chromatography with diode array detection. It was observed that the polyphenolic composition in apple peel depended on varieties, whereas the main classes of phenolic compounds in apple pulp were flavan-3-ols and hydroxycinnamic acids in all cases, representing both together between 86 and 95% of total polyphenols assayed.

Chromatography, High Pressure Liquid↗