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Supplementation with a pine bark extract rich in polyphenols increases plasma antioxidant capacity and alters the plasma lipoprotein profile.

Pycnogenol (PYC), an extract of French maritime pine bark (Pinus pinaster), is a potent antioxidant with potential health benefits. Its bioavailabilty has previously been shown by urinary excretion studies of constituents and metabolites of PYC. The aim of this study was to test the effect of PYC supplementation on measures of oxidative stress and the lipid profile in humans. Twenty-five healthy subjects received PYC (150 mg/d) for 6 wk. Fasting blood was collected at baseline, after 3 and 6 wk of supplementation, and again after a 4-wk washout period. After 6 wk of supplementation with PYC, a significant increase in plasma polyphenol levels was detectable, which was reversed after the 4-wk washout phase. The antioxidant effect of PYC was demonstrated by a significant increase in oxygen radical absorbance capacity (ORAC) in plasma throughout the supplementation period (P < 0.05). The ORAC value returned to baseline after the 4-wk washout period. Moreover, in addition to its antioxidant effects, PYC significantly reduced LDL-cholesterol levels and increased HDL-cholesterol levels in plasma of two-thirds of the subjects. While the LDL changes reversed during washout, the HDL increase did not. There was no significant difference in LDL oxidizability or plasma lipid peroxides following PYC supplementation. Hence, following oral supplementation in humans, PYC significantly increases antioxidant capacity of plasma, as determined by ORAC, and exerts favorable effects on the lipid profile.

Administration, Oral↗

Inhibition of gastric H+,K(+)-ATPase and acid secretion by cassigarol A, a polyphenol from Cassia garrettiana Craib.

The effects of cassigarol A, a naturally occurring polyphenol, on gastric H+,K(+)-ATPase and gastric acid secretion were studied. Cassigarol A inhibited H+,K(+)-ATPase and K-stimulated p-nitrophenyl phosphatase from hog gastric mucosa with 50% inhibition of 1.2 x 10(-6) and 6.3 x 10(-6) M, respectively. The kinetic study showed that the inhibition of H+,K(+)-ATPase by cassigarol A was competitive with respect to ATP and non-competitive with respect to K+. Cassigarol A inhibited both H+,K(+)-ATPase-mediated proton transport and 2-deoxy-D-glucose-induced acid secretion. On the other hand, cassigarol A acetate, in which phenolic hydroxy groups are acetylated, was not effective in the inhibition of enzyme activity and acid secretion. These results indicate that cassigarol A is a potent inhibitor of gastric H+,K(+)-ATPase, that the anti-secretory activity of cassigarol A is related to the inhibition of H+,K(+)-ATPase and that an important moiety of cassigarol A in the interaction with the enzyme is the phenolic hydroxy groups.

Adenosine Triphosphatases↗

Antioxidative effect of polyphenol extract prepared from various Chinese teas.

METHODS. Twelve different types of Chinese teas, including green, semifermented, and black tea, were studied for their antioxidant activities and active components. Compositions of (-)-epicatechin, (-)-epigallocatechin, (-)-epicatechin gallate, (-)-epigallocatechin gallate, and gallic acid were identified by fast atom bombardment-mass spectrometry and high-performance liquid chromatography-mass spectrometry and quantified by high-performance liquid chromatography. Antioxidant activities in lard were measured by the Rancimat method. RESULTS. The results showed that both yields of polyphenol extract and antioxidant activities varied with different tea processing methods. It was found that (-)-epigallocatechin gallate, (-)-epigallocatechin, and (-)-epicatechin gallate inhibited soybean lipoxygenase at the IC50 values ranging from 10 to 20 microM.

Animals↗

Antimutagenic activity of green tea polyphenols.

For centuries green tea has been a widely consumed beverage throughout the world. It is known to contain a number of pharmacologically active compounds. In this study water extracts of green tea (WEGT) and their major constituents, green tea polyphenols (GTP), were examined for antimutagenic activity. WEGT and GTP were found to significantly inhibit the reverse mutation induced by benzo[alpha]pyrene (BP), aflatoxin B1 (AFB1), 2-aminofluorene, and methanol extracts of coal tar pitch in Salmonella typhimurium TA100 and/or TA98 in the presence of a rat-liver microsomal activation system. GTP also inhibited gene forward mutation in V79 cells treated with AFB1 and BP, and also decreased the frequency of sister-chromatid exchanges and chromosomal aberrations in V79 cells treated with AFB1. The addition of GTP during and after nitrosation of methylurea resulted in a dose-dependent inhibition of mutagenicity. Studies to define the mechanism of the antimutagenic activity of GTP suggest that it may affect carcinogen metabolism, DNA adduct formation, the interaction of ultimate carcinogen or the scavenging of free radicals.

Aflatoxin B1↗

Two rapid microscale procedures for isolation of total RNA from leaves rich in polyphenols and polysaccharides: application for sensitive detection of grapevine viroids.

Two rapid microscale procedures for isolation of total RNA from grapevine leaves are described. Homogenized leaf tissue is phenol extracted and polyphenols and polysaccharides are removed by either an aqueous two-phase system followed by DEAE-cellulose chromatography or by differential precipitation with the solvent 2-butoxyethanol. The resulting RNA, about 5-15 micrograms from 0.25 g grapevine leaves, is of high quality and can be employed for reverse transcription and polymerase chain reaction (PCR) amplification or Northern blot analysis to detect grapevine viroids or other RNAs. The two procedures are optimized for small-scale examination of many samples in a short time. Thus RNA from 12-24 leaf samples can be analyzed per day, including reverse transcription and PCR amplification. The application of these procedures led to the first detection of grapevine yellow speckle viroid 1 in German grapevines.

Base Sequence↗

Protective effects of green tea polyphenols administered by oral intubation against chemical carcinogen-induced forestomach and pulmonary neoplasia in A/J mice.

Our studies and others have shown the cancer chemopreventive effects of chronic administration of green tea in several animal tumor models. In this study, the administration of a polyphenolic fraction isolated from green tea (GTP) by oral intubation at a dose of 5 mg in 0.2 ml water 30 min prior to challenge with carcinogen, afforded significant protection against both diethylnitrosamine (DEN)- and benzo(a)pyrene (BP)-induced forestomach and lung tumorigenesis in A/J mice. The protective effects were evident by a decrease in numbers of tumors/mouse in GTP-fed groups compared to non GTP-fed controls. In the forestomach tumorigenesis protocol, GTP afforded 71 and 66% protection against, respectively DEN- and BP-induced tumor multiplicity. In the case of lung tumorigenesis protocol, however, the protective effects of GTP were 41 and 39%, respectively. Histological examination of forestomach tumors showed significantly lesser number of squamous cell carcinoma formation in GTP-fed groups of mice compared to carcinogen alone-treated controls. When pulmonary tumors were examined histologically, no adenocarcinomas were observed in GTP-fed groups compared to 15% mice with adenocarcinomas in DEN and BP alone-treated controls. The results of this study suggest that limited doses of GTP administration by gavage 30 min prior to carcinogen challenge may afford protection against carcinogen-induced tumorigenesis in internal body organs.

Administration, Oral↗

Chromatographic analysis of polyphenols of some Iberian Thymus.

A comparative chromatographic analysis of polyphenols of some Iberian Thymus species of the sections Pseudothymbra, Thymus and Serpyllum was carried out by means of HPLC and TLC, in order to investigate their pharmacological interest. The purified leaf extracts were analysed and compared with authentic samples of 15 flavones, one flavonol, four flavanones, two dihydroflavonols, and caffeic and rosmarinic acids. The taxa belonging to the section Pseudothymbra and some of those in the section Thymus showed the highest content of methoxylated flavonoids, which are known to have antispasmodic properties.

Chromatography, High Pressure Liquid↗

Improvement in bovine embryo production in vitro by treatment with green tea polyphenols during in vitro maturation of oocytes.

The present study examined the effect of green tea polyphenols (GTP) during in vitro maturation (IVM) of bovine oocytes on in vitro fertilization (IVF) parameters, intracellular glutathione (GSH) concentration and subsequent embryo development. Cumulus-oocyte complexes were aspirated from the ovaries derived from slaughterhouse and cultured in modified synthetic oviduct fluid (m-SOF) supplemented with 0-25 microM GTP for 24h. After IVM, cumulus-free oocytes were coincubated with frozen-thawed spermatozoa for 15-18 h. Putative embryos were transferred to m-SOF and cultured for 8 days (Experiment 1). In comparison with the absence of GTP, treatment with GTP at a concentration of 15 microM showed a significant increase in the proportion of pronuclear (PN) formation after sperm penetration (65% versus 80%, P<0.05). No significant differences in the rates of sperm penetration and polyspermic fertilization were found among treatments. The cleavage rate at 48 h of in vitro insemination showed no difference in oocytes matured with or without GTP. However, compared to no addition (23.5%), the presence of 15 and 20 microM GTP during IVM significantly (P<0.05) increased the proportion of blastocysts (38.1% and 36.4%) on day 9 of in vitro insemination. A further increase from 20 to 25 microM GTP reduced (P<0.05) the proportion of blastocysts. In Experiment 2, after IVM, oocytes were fixed to analyze the GSH concentration. Compared to no addition, a higher (P<0.05) level of GSH was found in oocytes matured with 15 microM GTP and compared with 15 microM GTP, GSH was low (P<0.05) at 20 and 25 microM GTP. The results suggest that at certain concentrations of GTP (15 microM) in IVM medium has beneficial effects on subsequent embryo development, and is correlated with intracellular GSH level in bovine oocytes.

Animals↗

The effect of acute red wine polyphenol consumption on postprandial lipaemia in postmenopausal women.

Postprandial lipoproteins are potentially atherogenic. The aim of this study was to elucidate whether acute consumption of red wine (RW) and dealcoholised red wine (DRW) regulates postprandial lipid and lipoprotein metabolism in 17 dyslipidaemic postmenopausal women. A mixed meal accompanied by either water, RW or DRW was consumed on three separate visits, in random order, 2 weeks apart. One fasting and 6 hourly postprandial blood samples were taken for lipid analysis. Results showed no significant quantitative changes in postprandial apolipoprotein (apo) B48 levels following the consumption of DRW or RW compared to water. However, qualitatively, DRW may reduce arterial exposure to apoB48-containing lipoproteins over the 6-h postprandial period measured. DRW consumption did not significantly change postprandial TG or insulin levels. A 35% (p = 0.02) increase in postprandial triglyceride (TG) levels and a 54% (p = 0.02) increase in insulin levels were observed following RW consumption, compared to water. In conclusion, acute DRW consumption had no effect on postprandial lipid and lipoprotein metabolism in dyslipidaemic postmenopausal women. However, the consumption of full-compliment RW exacerbated the postprandial lipaemic and insulin response over the 6-h period. Collectively, our findings suggest that neither polyphenols nor red wine reduce atherosclerotic risk by acutely modulating postprandial lipaemia over a 6-h period.

Arteriosclerosis↗

Suppressive effects of cacao liquor polyphenols (CLP) on LDL oxidation and the development of atherosclerosis in Kurosawa and Kusanagi-hypercholesterolemic rabbits.

We investigated the properties of cacao liquor polyphenols (CLP), which have an antioxidative effect on low-density lipoprotein (LDL) and an anti-atherosclerotic effect in the spontaneous familial hypercholesterolemic model, the Kurosawa and Kusanagi-hypercholesterolemic (KHC) rabbit. After 6 months of dietary administration of CLP at 1% (w/w) to the KHC rabbits, a higher total cholesterol concentration was observed in the treatment group compared to the control group. However, no other effects were noted in lipid profiles in plasma or lipoproteins. The plasma concentration of thiobarbituric acid reactive substances (TBARS), which is a lipid-peroxidation index, was significantly decreased 1 month after the start of CLP administration compared to that of the control group. The antioxidative effect of CLP on LDL was observed from 2 to 4 months of administration. The area of atherosclerotic lesions in the aorta in the CLP group (32.01+/-1.58%) was significantly smaller than that in the control group (47.05+/-3.29%), and the tissue cholesterol and TBARS concentrations were lower in the CLP group than in the control group. The anti-atherosclerotic effect of CLP was confirmed both rheologically and histopathologically. An in vitro study using KHC rabbit-derived LDL revealed that CLP significantly prolonged the lag time of LDL oxidation that was induced by a lipophilic azo-radical initiator, 2,2'-azobis(4-methoxy)-2,4-dimethylvaleronitrile (V-70), or Cu(2+) from a low concentration of 0.1 microg/mL. The antioxidative effect of CLP was superior to those of the well-known antioxidative substances, vitamin C, vitamin E and probucol. Therefore, CLP suppressed the generation of atherosclerosis, and its antioxidative effect appeared to have an important role in its anti-atherosclerotic activity.

Animals↗

Red wine polyphenols cause endothelium-dependent EDHF-mediated relaxations in porcine coronary arteries via a redox-sensitive mechanism.

Moderate consumption of wine is associated with cardiovascular protection most likely by increasing the endothelial formation of nitric oxide (NO). The present study investigated whether red wine polyphenolic compounds (RWPCs) increase the formation of endothelium-derived hyperpolarizing factor (EDHF) in arteries and, if so, to characterize the underlying mechanism. Porcine coronary artery rings were suspended in organ chambers for measurement of changes in isometric tension and membrane potential in the presence of indomethacin and N(omega)-nitro-L-arginine. RWPCs caused pronounced endothelium-dependent relaxations and hyperpolarizations, which were reduced by the combination of charybdotoxin plus apamin (two inhibitors of EDHF-mediated responses). Both responses to RWPCs were also reduced by antioxidants, membrane permeant analogues of superoxide dismutase, and diphenylene iodonium, an inhibitor of flavin-dependent enzymes. RWPCs induced the formation of superoxide in cultured endothelial cells. These findings demonstrate that RWPCs cause EDHF-mediated relaxations of coronary arteries, which are critically dependent on a redox-sensitive mechanism involving a flavin-dependent enzyme.

Animals↗

Resveratrol, a polyphenol found in grapes, suppresses oxidative damage and stimulates apoptosis during early colonic inflammation in rats.

Oxidative stress, neutrophil infiltration, proinflammatory cytokines and eicosanoid generation are clearly involved in the pathogenesis of intestinal bowel disease. Resveratrol, a polyphenolic compound found in grapes and wine, has been shown to have anti-inflammatory, antioxidant, antitumour and immunomodulatory activities, however, its effects on experimental colitis remain unknown. We have investigated the effects of resveratrol on the colon injury caused by intracolonic instillation of trinitrobenzenesulphonic acid (TNBS) in rats. We determined the production of prostaglandin (PG)E(2) and PGD(2) in colon mucosa and the expression of cyclo-oxygenases (COX)-1 and -2 immunohistochemically. The inflammatory response was assessed by histology and myeloperoxidase activity, as an index of neutrophil infiltration. Interleukin-1 beta production, histological and histochemical analysis of the lesions were also carried out. Finally, since resveratrol has been found to modulate apoptosis we intended to elucidate its effects on colonic mucosa under early acute inflammatory conditions. Resveratrol (5-10mg/kg/day) significantly reduced the degree of colonic injury, the index of neutrophil infiltration and the levels of the cytokine. Resveratrol did not revert the increased PGE(2) levels but produced a significant fall in the PGD(2) concentration. Compared with inflamed colon, no changes in staining for COX-1 were observed in colon of resveratrol and TNBS-treated rats. In contrast, COX-2 expression was decreased. Furthermore, resveratrol enhanced apoptosis compared with already high level induced by TNBS. In conclusion, resveratrol reduces the damage in experimentally induced colitis, alleviates the oxidative events and stimulates apoptosis.

Animals↗

Dietary polyphenols protect dopamine neurons from oxidative insults and apoptosis: investigations in primary rat mesencephalic cultures.

Naturally occurring polyphenols have the potential to prevent oxidative damage in various pathophysiological conditions. Various members of the flavonoid family were investigated to determine if they could protect mesencephalic dopamine (DA) neurones from injury and reduce apoptosis produced by oxidative stressors. Primary mesencephalic cultures were sensitive to oxidative insults (hydrogen peroxide, 4-hydroxynonenal, rotenone, 6-hydroxydopamine and N-methyl-4-phenyl-1,2,3,6-tetrahydropyridinium hydrochloride (MPP+)) which produced concentration-dependent decreases in cellular viability across an apoptotic-necrotic continuum of injury. Flavonoids (catechin, quercetin, chrysin, puerarin, naringenin, genestein) protected mesencephalic cultures from injury by MPP+, which was shown by DNA fragmentation studies and tyrosine hydroxylase (TH) immunocytochemistry of DA neurones to occur by apoptosis. Catechin also reduced injury produced by hydrogen peroxide, 4-hydroxynonenal, rotenone and 6-hydroxydopamine as shown by increases in cellular viability and [3H]DA uptake. When the neuroprotection of catechin against MPP+-induced injury was compared to that produced by the caspase-3 inhibitor, Z-DVED-FMK, both reduced DNA fragmentation and the injury patterns of TH-positive neurones. These data demonstrate the neuroprotective abilities of flavonoids which are able to attenuate the apoptotic injury of mesencephalic DA neurones. Since these DA neurones are under oxidative stress in Parkinsonism, our findings suggest that flavonoids could provide benefits along with other anti-oxidant therapies in Parkinson's disease.

Animals↗

The grape-derived polyphenol resveratrol differentially affects epidermal and platelet-derived growth factor signaling in human liver myofibroblasts.

The grape-derived polyphenol resveratrol is anti-proliferative for human liver myofibroblasts, which may be beneficial for the treatment of liver fibrosis. However, its mechanism of action is ill understood. Here, we have studied how resveratrol interfered with signaling pathways used by epidermal or platelet-derived growth factors to induce the proliferation of these cells. We found that resveratrol inhibited epidermal growth factor or platelet-derived growth factor-induced DNA synthesis. Resveratrol did not, however, decrease epidermal growth factor receptor autophosphorylation or activation of extracellular regulated kinases, but strongly inhibited the phosphorylation of Akt and of its substrate forkhead related transcription factor. This suggested that resveratrol inhibited epidermal growth factor-induced mitogenic signaling through inhibition of the phosphatidylinositol 3-kinase /Akt pathway. The phosphatidylinositol 3-kinase inhibitor LY 294002, also, inhibited epidermal growth factor-dependent DNA synthesis and Akt phosphorylation but did not decrease extracellular regulated kinases phosphorylation. In contrast, resveratrol inhibited platelet-derived growth factor-stimulated receptor autophosphorylation and every subsequent signaling step. Resveratrol did not directly inhibit phosphatidylinositol 3-kinase activity measured on immunoprecipitates from epidermal growth factor-stimulated myofibroblasts, but it strongly reduced the autophosphorylation of the phosphatidylinositol 3-kinase downstream target phospho-inositide-dependent kinase-1 that phosphorylates Akt. We, thus, show that resveratrol has growth factor-specific effects: it inhibits platelet-derived growth factor signaling via reduced receptor activation, whereas it reduces epidermal growth factor-dependent DNA synthesis via inhibition of the phosphatidylinositol 3-kinase/Akt pathway, possibly through inhibition of phospho-inositide-dependent kinase-1 activity.

Cell Proliferation↗

Structural requirements for stabilization of vascular elastin by polyphenolic tannins.

Elastin-associated degeneration and calcification are potential causes of long-term failure of glutaraldehyde (Glut) fixed tissue bioprostheses used in cardiovascular surgery. This vulnerability may be attributed to the inability of Glut to cross-link and adequately protect vascular elastin from enzymatic attack. Tannic acid (TA), a poly galloyl glucose (Glc), is compatible with Glut fixation, binds to vascular elastin, improves resistance to degradation and reduces in vivo calcification. While these results provided evidence of a beneficial interaction between elastin and TA, the nature and mechanisms of these interactions are unclear; moreover, TA-elastin binding exhibits a partial instability after long-term interaction with vascular elastin which could contribute to issues of implant toxicity. In present studies, we used resistance to elastase, mechanical properties, and cell viability assays to evaluate the elastin-stabilizing potential and cytotoxicity of TA derivatives and individual TA components such as acetylated TA (AcTA), pentagalloylglucose (PGG), free gallic acid (Gall) and Glc. Our comparative study demonstrates that polyphenolic hydroxyl groups are the main structural groups essential to the interaction between TA and elastin. Furthermore, we show that PGG, the core structure of TA, possesses the same unique elastin-stabilizing qualities of TA, yet it is much less cytotoxic than TA and thus could be potentially useful as an elastin-stabilizing agent for cardiovascular bioprostheses.

Animals↗

Characterization of a fungal strain isolated from a polyphenol polluted site.

A group of fungal strains were isolated from a polyphenol polluted soil, taken from an olive oil processing plant in Attica, Greece. The fungi were tested for their ability to decolorize a polyaromatic dye Poly R-478, which was used as a model compound to test their ligninolytic activities. The strain K1.1 decolorized efficiently the dye on agar plates and was further studied. PCR amplification of the internal transcribed spacer (ITS) region of the ribosomal RNA genes from the genomic DNA isolated from mycelium grown in liquid culture resulted in amplified fragments. Via BLASTN search, the length of a 773 base pairs was identified as the basidiomycetes Coprinellus xanthothrix. The growth rates and the tolerance of the fungus were compared on solid media, containing four different concentrations of pentachlorophenol. Extracellular enzyme activities (lignin peroxidase, manganese peroxidase and laccase) were determined in defined liquid medium. The isolate expressed laccase and manganese peroxidase but not lignin peroxidase. The removal of the dye was also estimated in liquid medium. The fungus showed biosorption and biotransformation as removal mechanisms.

Adsorption↗

4-Alkyliden-beta-lactams conjugated to polyphenols: synthesis and inhibitory activity.

A series of compounds combining the beta-lactam and polyphenol scaffold have been prepared and evaluated for inhibition of human leukocyte elastase and matrix metallo-proteases MMP-2 and MMP-9. The design of these compounds has been based on the 'overlapping-type' strategy where two pharmacophores are linked in a single molecule. The most powerful compound against elastase was an N-galloyl-4-alkyliden beta-lactam, [3-[1-(tert-butyl-dimethyl-silanyloxy)-ethyl]-4-oxo-1-(3,4,5-tris-benzyloxy-benzoyl)-azetidin-2-ylidene]-acetic acid ethylester, with an IC50 of 0.5 microM; while the most powerful against MMP-2 was a 4-alkyliden beta-lactam arylated on the C-3 hydroxy side chain (3,5-bis-benzyloxy-4-hydroxy-benzoic acid 1-(2-benzyloxycarbonylmethylene-4-oxo-azetidin-3-yl)-ethyl ester) with an IC50 of 4 microM. Of the total 35 compounds tested, high levels of inhibition of elastase and of MMPs were separately exerted by distinct molecules.

Cell Line, Tumor↗