gallocatechin-3-gallate and epicatechin-gallate

High performance liquid chromatography (HPLC) for catechins and related compounds in

Topics: Caffeine; Camellia sinensis; Catechin; Chemistry Techniques, Analytical; Chromatography, High Pressure Liquid; Gallic Acid; Plant Extracts; Plant Leaves; Reference Standards; Thailand; Validation Studies as Topic

Xinyang Maojian tea is a kind of famous roasted green tea produced in the middle of China. Ultra-high performance liquid chromatography-quadrupole time of flight-mass spectrometry (UHPLC-Q-TOF/MS)-based metabolomics coupled with multivariate statistical analyses, including principal component analysis (PCA) and hierarchical cluster analysis (HCA), were carried out in XMMJTs collected from Luoshan, Shangcheng, and Shihe Counties, respectively. Additionally, seven catechins, four flavonoids, two purine alkaloids, and gallic acid contents were determined by HPLC. Differential metabolites were selected by p-value <0.05, and fold change >1.50 or < 0.66 among 745 detected metabolites in metabolomics analysis. The results showed significant (p < 0.05) differences of three catechins including (-)-epicatechin, (-)-epicatechin gallate, and (-)-gallocatechin gallate, four flavonoids (i.e. quercetin, kaempferol, myricetin, and rutin), and theobromine among three various regions, and significant (p < 0.05) differences of (-)-epicatechin gallate, (-)-epigallocatechin, (+)-catechin, gallic acid, and kaempferol between Shuchazao and Group cultivar. The HCA showed that, except for two samples (i.e. LS 2 and SH 2) of Shuchazao cultivar clustered together, others could be clustered completely according to production place. The 63 relevant differential metabolites could achieve the purpose of region identification through PCA. Kyoto encyclopedia of genes and genomes (KEGG) metabolic pathway analysis elaborated the impact of geographical origin and tea cultivar on physiological metabolism in tea tree. PRACTICAL APPLICATION: Ultra-high performance liquid chromatography-quadrupole time of flight-mass spectrometry (UHPLC-Q-TOF/MS)-based liquid chromatography-tendem mass spectrometry (LC-MS/MS) metabolomics coupled with multivariate statistical analyses, such as principal component analysis (PCA) and hierarchical cluster analysis (HCA), revealed 63 differential metabolites related to production place, which contributed to the region identification of Xinyang Maojian teas.

Topics: Camellia sinensis; Catechin; China; Chromatography, High Pressure Liquid; Chromatography, Liquid; Flavonoids; Gallic Acid; Metabolomics; Multivariate Analysis; Plant Leaves; Tandem Mass Spectrometry; Tea

Structural investigations, based on density functional theory (DFT) calculations, are performed on tea catechins, including 4-aminobutyric acid (GABA), L-theanine (Thea), caffeine (CAF), theobromine (TB), theophylline (TP), catechin (C), epicatechin (EC), gallocatechin (GC), epigallocatechin (EGC), catechin gallate (CG), epicatechin gallate (ECG), gallocatechin gallate (GCG) and epigallocatechin gallate (EGCG). With an identified lowest energy conformer of investigated molecules, FTIR and FT-Raman spectra have been assigned according to DFT calculations in the way of B3LYP/6-31 + G (d, p). Normal spectra of these catechin powders are also measured by Raman spectrometers. There is a kind of everlasting correlation between experimental results and theoretical data. And our research has also obtained a clear evidence for reliable assignments of vibrational bands, bringing great feasibility to the rapid tea catechin detection.

Topics: Catechin; Spectroscopy, Fourier Transform Infrared; Spectrum Analysis, Raman; Tea

Tea catechins, the main bioactive polyphenols in green tea, are well known for their health promoting effects. Previous studies have shown that gallocatechin-3-gallate (GCG), epigallocatechin-3-gallate (EGCG) and epicatechin-3-gallate (ECG) exerted strong inhibitory effects on mushroom tyrosinase activity in vitro, whilst EGCG inhibited melanogenesis in vivo, yet the underlying mechanisms are not entirely clear. In this study, we (i) evaluated and compared the inhibitory effects of the main tea catechins (GCG, EGCG, and ECG) on melanogenesis in B16F10 melanoma cells, and (ii) explain the underlying mechanisms. The results showed that the tea catechins significantly suppressed tyrosinase activity and melanin synthesis in B16F10 cells, where the effects of ECG > EGCG > GCG. Interestingly, the inhibitory effects of the catechins were stronger than those of arbutin (AT), a well-known depigmenting agent. Moreover, GCG, EGCG, and ECG regulated the melanogenesis of B16F10 cells through the cAMP/CREB/MITF pathway. These results revealed catechins could be used as anti-melanogenic agents to protect cells from abnormal melanogenesis.

Topics: Animals; Catechin; Cell Line, Tumor; Cyclic AMP; Cyclic AMP Response Element-Binding Protein; Down-Regulation; Melanins; Melanoma, Experimental; Mice; Microphthalmia-Associated Transcription Factor; Molecular Structure; Monophenol Monooxygenase; Signal Transduction; Tea

Many studies have shown that flavan-3-ols inhibit mammalian alpha-amylases but the published IC

Topics: alpha-Amylases; Amylose; Animals; Catechin; Digestion; Flavonoids; Hydrolysis; Inhibitory Concentration 50; Models, Molecular; Oligosaccharides; Starch; Swine; Tea

Oolong tea is an important member in tea family, which claims for various health benefits such as preventing obesity and improving lipid metabolism. In this work, using pancreatic lipase (PL) functionalised magnetic nanoparticles (PL-MNPs) as solid phase extraction absorbent in combination with ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS), we developed a method for rapid screening and identification of lipase inhibitors from oolong tea. Three PL ligands were selectively extracted and identified as (-)-epigallocatechin-3-O-gallate (EGCG), (-)-gallocatechin-3-O-gallate (GCG) and (-)-epicatechin-3-O-gallate (ECG). Their lipase inhibitory activities were significantly higher than those non-ligands. Structure-activity analysis revealed that the presence of a galloyl moiety in the structure was required for binding to PL-MNPs, and therefore, exhibiting a strong inhibition on the enzyme. Taking advantages of the specificity in enzyme binding and the convenience of magnetic separation, this method has great potential for fast screening of lipase inhibitors from natural resources.

Topics: Catechin; Chromatography, High Pressure Liquid; Enzyme Inhibitors; Lipase; Magnetite Nanoparticles; Plant Extracts; Tandem Mass Spectrometry; Tea

The effects of green tea catechins on glucose-stimulated insulin secretion (GSIS) were investigated in the β-cell line INS-1D. Epigallocatechin gallate (EGCG) at 10 µM or gallocatechin gallate (GCG) at 30 µM caused significant inhibitory effects on GSIS, and each of these at 100 µM almost abolished it. In contrast, epicatechin (EC) or catechin (CA) had no effect on GSIS at concentrations up to 100 µM. We thus investigated the structure-activity relationship by using epigallocatechin (EGC) and gallocatechin (GC) containing a trihydroxyl group in the B-ring, and epicatechin gallate (ECG) and catechin gallate (CG) containing the gallate moiety. EGC, GC, and ECG caused an inhibition of GSIS, although significant effects were obtained only at 100 µM. At this concentration, EGC almost abolished GSIS, whereas GC and ECG partially inhibited it. In contrast, CG did not affect GSIS at concentrations up to 100 µM. EGCG also abolished the insulin secretion induced by tolbutamide, an ATP-sensitive K(+) channel blocker, and partially inhibited that induced by 30 mM K(+). Moreover, EGCG, but not EC, inhibited the oscillation of intracellular Ca(2+) concentration induced by 11.1 mM glucose. These results suggest that some catechins at supraphysiological concentrations have inhibitory effects on GSIS, the potency of which depends on their structure; the order of potency was EGCG>GCG>EGC>GC≈ECG. The inhibitory effects seem to be mediated by the inhibition of voltage-dependent Ca(2+) channels, which is caused, at least in part, by membrane hyperpolarization resulting from the activation of K(+) channels.

Topics: Calcium; Calcium Channels; Catechin; Glucose; Insulin; Insulin Secretion; Insulin-Secreting Cells; Structure-Activity Relationship; Tea

Skin hyper-pigmentation is a condition initiated by the overproduction of melanin existing in the melanocytes. Melanin pigment is responsible for the colour of skin in humans. It is formed through a series of oxidative reactions involving the amino acid tyrosine in the presence of the key enzyme tyrosinase. In continuation with our efforts to identify tyrosinase inhibitors from plants sources, the methanol extract from leaf, bark and fruit of Ceratonia siliqua were screened for tyrosinase inhibition and diphenolase activity. The bark extract exhibited significant inhibition on mushroom tyrosinase using L-tyrosine as a substrate and showed diphenolase activity. The extract further significantly lowered tyrosinase mRNA levels in B16-F10 mouse melanocytes. Bioassay-guided fractionation led to the isolation of six compounds. Compounds (-)-epicatechin-3-O-gallate, 1,2,3,6-tetra-O-galloyl-ß-D-glucose and gallocatechin-3-O-gallate showed tyrosinase inhibitions with the IC50 values of 27.52, 83.30 and 28.30 µg/mL, respectively. These compounds also exhibited L-DOPA activities with IC50 values of >200, 150 and 200 µg/mL, respectively. A clinical study was conducted using 20 volunteers in a patch testing trial for irritancy potential and skin depigmentation. The clinical results showed the sample to be non-irritant with irritancy potential of -34.21 and depigmentation trial showed an improvement in the even skin tone of UV induced pigmentation at 3% after 28 days of application.

Topics: Agaricales; Animals; Catechin; Enzyme Inhibitors; Fabaceae; Glucose; Humans; Levodopa; Melanins; Melanocytes; Mice; Monophenol Monooxygenase; Plant Extracts; Skin

The aim of this study was to evaluate the effects of green tea Camellia sinensis extract on proinflammatory molecules and lipolytic protein levels in adipose tissue of diet-induced obese mice. Animals were randomized into four groups: CW (chow diet and water); CG (chow diet and water + green tea extract); HW (high-fat diet and water); HG (high-fat diet and water + green tea extract). The mice were fed ad libitum with chow or high-fat diet and concomitantly supplemented (oral gavage) with 400 mg/kg body weight/day of green tea extract (CG and HG, resp.). The treatments were performed for eight weeks. UPLC showed that in 10 mg/mL green tea extract, there were 15 μg/mg epigallocatechin, 95 μg/mg epigallocatechin gallate, 20.8 μg/mg epicatechin gallate, and 4.9 μg/mg gallocatechin gallate. Green tea administered concomitantly with a high-fat diet increased HSL, ABHD5, and perilipin in mesenteric adipose tissue, and this was associated with reduced body weight and adipose tissue gain. Further, we observed that green tea supplementation reduced inflammatory cytokine TNFα levels, as well as TLR4, MYD88, and TRAF6 proinflammatory signalling. Our results show that green tea increases the lipolytic pathway and reduces adipose tissue, and this may explain the attenuation of low-grade inflammation in obese mice.

Topics: Adiponectin; Animals; Catechin; Chromatography, High Pressure Liquid; Diet, High-Fat; Enzyme-Linked Immunosorbent Assay; Interleukin-10; Lipolysis; Mice; Myeloid Differentiation Factor 88; Obesity; Tea; TNF Receptor-Associated Factor 6; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

this study examined the effects of a dentifrice containing green tea catechins on gingival oxidative stress and periodontal inflammation using a rat model.. twenty-four male Wister rats were randomly divided into four groups. The first group (Control group) received no treatment for 8 weeks. Periodontal inflammation was induced in the second group for 8 weeks. Periodontal inflammation was induced in the last two groups for 8 weeks and dentifrices with or without green tea catechins were topically applied to the gingival sulcus daily for 4 weeks prior to the end of the experimental period.. rats that had experimental periodontal inflammation showed apical migration of the junctional epithelium, alveolar bone loss and inflammatory cell infiltration in the connective tissue subjacent to the junctional epithelium at 8 weeks, whilst the control group showed no pathologic changes. Topical application of a green tea catechin-containing dentifrice reduced inflammatory cell infiltration in the periodontal lesions to a greater degree than the control dentifrice at 8 weeks. The gingiva in which green tea catechin-containing dentifrice was applied also showed a lower level of expression of hexanoyl-lysine (a marker of lipid peroxidation), nitrotyrosine (a marker of oxidative protein damage), and tumour necrosis factor-α (an indicator of pro-inflammatory cytokines) at 8 weeks compared to gingiva in which the control dentifrice was applied.. adding green tea catechins to a dentifrice may contribute to prevention of periodontal inflammation by decreasing gingival oxidative stress and expression of pro-inflammatory cytokines.

Topics: Alveolar Bone Loss; Animals; Antioxidants; Camellia sinensis; Catechin; Connective Tissue; Dentifrices; Disease Models, Animal; Epithelial Attachment; Gingiva; Gingival Recession; Lipid Peroxidation; Lysine; Male; NF-kappa B; Oxidative Stress; Periodontitis; Random Allocation; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha; Tyrosine

Moderate red wine consumption has been correlated with lower incidences of cardiovascular diseases, inflammation, and metabolic diseases such as type 2 diabetes, obesity, and high blood pressure. We studied binding of ligands from different wines to the peroxisome proliferator-activated receptor γ (PPARγ), a key factor in glucose and lipid metabolism. Ellagic acid and epicatechin gallate (ECG) were identified by gas chromatography and mass spectroscopy in the most active wine fractions. They had an affinity to PPARγ similar to that of the standard pharmaceutical agent rosiglitazone, which is used for the treatment of type 2 diabetes. The IC(50) values of ellagic acid and ECG were 5.7 × 10(-7) M and 5.9 × 10(-7) M, respectively. All of the red wines had affinities for PPARγ equivalent to concentrations of rosiglitazone ranging from 52-521 μM. One hundred milliliters of the tested red wines was equivalent to approximately 1.8-18 mg of rosiglitazone. This volume contained an activity equivalent of at least a quarter of (and up to four times) the daily dose of this potent anti-diabetes drug. The ameliorating effects of red wine on metabolic diseases may be partially explained by the presence of PPARγ ligands.

Topics: Binding, Competitive; Cardiovascular Diseases; Catechin; Coumaric Acids; Diabetes Mellitus, Type 2; Ellagic Acid; Flavonoids; Gas Chromatography-Mass Spectrometry; Humans; Hypoglycemic Agents; Ligands; Models, Chemical; Phenols; Plant Extracts; Polyphenols; PPAR gamma; Rosiglitazone; Spectroscopy, Fourier Transform Infrared; Thiazolidinediones; Wine

Four catechins with the galloyl moiety, including catechin gallate (CG), epigallocatechin gallate (EGCG), gallocatechin gallate (GCG), and epicatechin gallate (ECG), were found to inhibit HIV-1 integrase effectively as determined by our ELISA method. In our docking study, it is proposed that when the HIV-1 integrase does not combine with virus DNA, the four catechins may bind to Tyr143 and Gln148, thus altering the flexibility of the loop (Gly140-Gly149), which could lead to an inhibition of HIV-1 integrase activity. In addition, after combining HIV-1 integrase with virus DNA, the four catechins may bind between the integrase and virus DNA, consequently, disrupt this interaction. Thus, the four catechins may reduce the activity of HIV-1 integrase by disrupting its interaction with virus DNA. The four catechins have a highly cooperative inhibitory effect (IC₅₀=0.1 μmol/L). Our study suggests that catechins with the galloyl moiety could be a novel and effective class of HIV-1 integrase inhibitors.

Topics: Acquired Immunodeficiency Syndrome; Anti-HIV Agents; Catechin; DNA, Viral; Enzyme-Linked Immunosorbent Assay; HIV Integrase; HIV Integrase Inhibitors; HIV-1; Humans; Pyrrolidinones; Raltegravir Potassium

A new on-line preconcentration technique called borate complexation-assisted field-enhanced sample injection (BCA-FESI) was proposed for preconcentrating cis-diol-containing compounds (CDCCs) in capillary electrophoresis (CE). The principle relies on amplification of the difference in the electrophoretic mobilities of CDCC in sample matrix and background electrolyte (BGE) through complexation of CDCC with borate in a sample matrix of basic pH and dissociation of the complex in a BGE of acidic pH. Meanwhile, CDCC and borate ions electro-injected into the capillary are finally in neutral state, which maintains the pre-filled low conductivity zone and thus allows for longer injection time. With catechol as a test compound, the principle and effectiveness of BCA-FESI was verified. As compared to conventional sample injection, BCA-FESI allowed for sensitivity enhancement of 1850-fold. The established method was further evaluated with three catechins, including (-)-epicatechin gallate (ECG), (-)-gallocatechin gallate (GCG), and (-)-epigallocatechin (EGC), in a standard mixture of trace content. The limit of detection (LOD) was found to be 1.4, 3.8, 17.5 nM (S/N=3) for ECG, GCG, EGC, respectively. Finally, the BCA-FESI method was applied to a real sample of diluted tea beverage, in which the three catechins were detected.

Topics: Algorithms; Borates; Catechin; Catechols; Electrolytes; Electrophoresis, Capillary; Hydrogen-Ion Concentration; Kinetics; Molecular Structure

Inclusion complexes of (-)-epicatechin gallate (ECg) as well as (+)-gallocatechin gallate (GCg) and beta-cyclodextrin (beta-CD) in an aqueous solution were investigated using several NMR techniques and a computational method. ECg and EGCg formed a 1:1 complex with beta-CD, in which the A ring and a portion of the C ring were included from the wide secondary hydroxyl group side of the beta-CD cavity, and the B and B' rings were left outside the cavity. GCg formed a 1:2 complex with beta-CD, in which the A and B rings of GCg were included by two molecules of beta-CD. The difference between the two modes of inclusion of the 1:1 complex of ECg, EGCg.beta-CD and the 1:2 complex of GCg.beta-CD might have resulted from the size of the space between the B and B' rings in aqueous solution. As a result of nuclear Overhauser effect (NOE) experiments, GCg was considered to have a large enough space between the B and B' rings to include the B ring in the beta-CD cavity; on the other hand, ECg and EGCg have no such large space.

Topics: Catechin; Magnetic Resonance Spectroscopy; Molecular Conformation; Solvents; Stereoisomerism; Water

Here, we investigated the structure-activity relationship of major green tea catechins and their corresponding epimers on cell-surface binding and inhibitory effect on histamine release. Galloylated catechins; (-)-epigallocatechin-3-O-gallate (EGCG), (-)-gallocatechin-3-O-gallate (GCG), (-)-epicatechin-3-O-gallate (ECG), and (-)-catechin-3-O-gallate (CG) showed the cell-surface binding to the human basophilic KU812 cells by surface plasmon resonance analysis, but their non-galloylated forms did not. Binding activities of pyrogallol-type catechins (EGCG and GCG) were higher than those of catechol-type catechins (ECG and CG). These patterns were also observed in their inhibitory effects on histamine release. Previously, we have reported that biological activities of EGCG are mediated through the binding to the cell-surface 67kDa laminin receptor (67LR). Downregulation of 67LR expression caused a reduction of both activities of galloylated catechins. These results suggest that both the galloyl moiety and the B-ring hydroxylation pattern contribute to the exertion of biological activities of tea catechins and their 67LR-dependencies.

Topics: Anti-Allergic Agents; Basophils; Catechin; Cell Line, Tumor; Glycosylation; Histamine Release; Humans; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Molecular Structure; Receptors, Laminin; Structure-Activity Relationship; Tea

Uniformly-sized, molecularly imprinted polymers (MIPs) for (-)-epigallocatechin gallate (EGCg), -epicatechin gallate (ECg) and -gallocatechin gallate (GCg) were prepared by a multi-step swelling and polymerization method using 2-vinylpyridine as a functional monomer, ethylene glycol dimethacrylate as a cross-linker and cyclohexanol as a porogen. Molecular recognition abilities of the obtained MIPs were evaluated in liquid chromatography using a mixture of ethanol and water, or ethanol as the eluent. Each MIP gave the highest molecular recognition ability for the respective template molecule. In addition, (-)-EGCg and -ECg had the same configuration (2R,3R) at positions 2 and 3, and therefore resulting in high cross reactivity each other. However, (-)-GCg, which has different configuration at position 2 with (-)-EGCg and -ECg, showed low cross reactivity with them. On the other hand, those MIPs showed no molecular recognition against (-)-epigallocatechin and -epicatechin, which have no gallate group at position 3. These results indicate that the MIPs prepared can recognize configuration at position 2 and a gallate group at position 3. Furthermore, the MIP for (-)-GCg could be successfully used for isolating (-)-EGCg and -ECg from green tea extract.

Topics: Catechin; Chromatography, High Pressure Liquid; Tea

Attomole quantities of catechins were determined by a capillary liquid chromatography system with electrochemical detection (CLC-ECD) and the system is applied to the determination of catechins in human plasma. The eight catechins: catechin (C), epicatechin (EC), gallocatechin (GC), epigallocatechin (EGC), catechin gallate (Cg), epicatechin gallate (ECg), gallocatechin gallate (GCg), and epigallocatechin gallate (EGCg), were separated within 10 min using a capillary column (0.2 mm i.d.) and a mobile phase of phosphoric acid (85%)-methanol-water (0.5:27.5:72.5, v/v/v), and were detected at +0.85 V vs. Ag/AgCl. Peak heights were found to be linearly related to the amount of catechins injected, from 200 amol to 500 fmol (r > 0.998). The detection limits of the catechins were 61 amol for EGC, 75 amol for EC, 54 amol for GC, 61 amol for C, 67 amol for GCg, 75 amol for EGCg, 75 amol for ECg and 89 amol for Cg (S/N = 3). Because the present method is highly sensitive and allows facile pretreatment for plasma sample, the time courses of concentrations of catechins (GCg, EC, EGCg, ECg, and Cg) and their conjugates in human plasma obtained from a 10 microl plasma sample after ingestion of green tea could be determined.

Topics: Catechin; Chromatography, Liquid; Electrochemistry; Humans; Methanol; Phosphoric Acids; Reproducibility of Results; Sensitivity and Specificity; Tea; Time Factors; Water

The purpose of this study was to examine the effects of catechins and their derivatives on the activities of Arg-gingipain (Rgp) and Lys-gingipain (Kgp) in Porphyromonas gingivalis. Catechin derivatives, which included (-)-epigallocatechin gallate, (-)-epicatechin gallate, (-)-gallocatechin gallate, and (-)-catechin gallate, significantly inhibited the Rgp activity. The 50% inhibitory concentrations (IC50s) of these catechin derivatives for Rgp ranged from 3 to 5 microm. While (-)-epigallocatechin and (-)-gallocatechin moderately inhibited Rgp activity (IC50s, 20 microm), (-) -epicatechin, (+)-catechin, and gallic acid were not effective, with IC50s greater than 300 microm. Further, some of the catechin derivatives tested also inhibited the Kgp activity, though to a lesser extent than inhibition of the Rgp activity. These findings suggest that green tea catechins may have the potential to reduce periodontal breakdown resulting from the potent proteinase activity of P. gingivalis.

Topics: Adhesins, Bacterial; Catechin; Cysteine Endopeptidases; Cysteine Proteinase Inhibitors; Gingipain Cysteine Endopeptidases; Hemagglutinins; Humans; Plant Extracts; Porphyromonas gingivalis; Tea

The anticancer properties of tea catechins are most frequently attributed to the principal catechin (-)-epigallocatechin-3-gallate (EGCg). Efficacy was evaluated using growth of cultured HeLa cells and inhibition of the enzymatic activity of a putative cell surface tea target enzyme, a cancer-associated cell surface-located NADH oxidase (ECTO-NOX) designated tNOX. The amounts of EGCg required to inhibit by both criteria was reduced 10 times by combination with inactive catechins such as (-)-epicatechin (EC), (-)-epigallocatechin (EGC) or (-)-epicatechin-3-gallate (ECG). Various synthetic mixtures based on purified catechins and decaffeinated tea extracts treated enzymatically to reduce the ester bond-containing catechins varying in EGCg content from 0.065 to 40% were of comparable efficacy to decaffeinated green tea extracts as long as EGCg was present and the ratio of total catechins to EGCg + EGC was about 1.5. Such mixtures appear to offer potential cancer protection and therapeutic advantages over those of EGCg alone through lowered toxicity of the mixture to normal cells and for more efficient blood delivery of orally-administered catechins to a tumour site.

Topics: Antineoplastic Agents; Antioxidants; Beverages; Catechin; Cell Division; Drug Synergism; HeLa Cells; Humans; Microscopy, Fluorescence; NADH, NADPH Oxidoreductases; Plant Extracts; Structure-Activity Relationship; Tea; Time Factors

Recent studies have shown that lead causes oxidative stress by inducing the generation of reactive oxygen species (ROS) and reducing the antioxidant defense system of cells, which suggests that antioxidants may play an important role in the treatment of lead poisoning. The present study was designed to elucidate whether tea catechins had any protective effects on altered oxidative stress parameter in PC12 cells exposed to lead. The experimental results showed that lead decreased PC12 cell viability and induced a rapid elevation of [Ca(2+)](i), which was followed by an accumulation of ROS and a decrease of mitochondrial membrane potential (MMP). Treatment by tea catechins significantly increased cell viability, decreased intracellular Ca(2+) levels and ROS formation, and improved MMP in PC12 cells exposed to lead. The galloylated catechins showed a greater effect on ROS formation and mitochondrial dysfunction than that of nongalloylated catechins, which was similar to the result of their scavenging ability on free radical. In view of the time course of ROS formation and mitochondrial dysfunction and their correlation, our results also suggested that the beneficial effects of tea catechins on MMP are related, at least in part, to its ability to scavenge ROS in PC12 cells exposed to 100 microM Pb(2+). The present results suggest that tea catechins supplementation may play a role for modulating oxidative stress in PC12 cells exposed to lead.

Topics: Animals; Calcium; Catechin; Cell Death; Cell Survival; Dose-Response Relationship, Drug; Lead; Membrane Potentials; Mitochondria; Oxidative Stress; PC12 Cells; Rats; Reactive Oxygen Species; Tea

It has been known that tea catechins, (-)-epicatechin (1), (-)-epigallocatechin (2), (-)-epicatechin gallate (3), and (-)-epigallocatechin gallate (4) are epimerized to(-)-catechin (5), (-)-gallocatechin (6), (-)-catechin gallate (7), and (-)-gallocatechin gallate (8), respectively, during retort pasteurization. We previously reported that tea catechins, mainly composed of 3 and 4, effectively inhibit cholesterol absorption in rats. In this study, the effect of heat-epimerized catechins on cholesterol absorption was compared with tea catechins. Both tea catechins and heat-epimerized catechins lowered lymphatic recovery of cholesterol in rats cannulated in the thoracic duct and epimerized catechins were more effective than tea catechins. The effect of purified catechins on micellar solubility of cholesterol was examined in an in vitro study. The addition of gallate esters of catechins reduced micellar solubility of cholesterol by precipitating cholesterol from bile salt micelles. Compounds 7 and 8 were more effective to precipitate cholesterol than 3 and 4, respectively. These observations strongly suggest that heat-epimerized catechins may be more hypocholesterolemic than tea catechins.

Topics: Animals; Catechin; Cholesterol; Hot Temperature; Intestinal Absorption; Male; Micelles; Rats; Rats, Sprague-Dawley; Solubility; Tea

The recent development of electrospray ionization mass spectrometry (ESI-MS) has allowed its use to study molecular interactions driven by non-covalent forces. ESI-MS has been used to detect non-covalent complexes between proteins and metals, ligands and peptides and interactions involving DNA, RNA, oligonucleotides and drugs. Surprisingly, the study of the interaction between polyphenolic molecules and peptides/proteins is still an area where ESI-MS has not benefited. With regard to the important influence of these interactions in the biological and food domains, ESI-MS was applied to the detection and the characterization of soluble polyphenol-peptide complexes formed in model solution. The ability to observe and monitor the weak interactions involved in such macromolecular complexation phenomena was demonstrated for monomeric and dimeric flavonoid molecules (catechin-derived compounds) largely encountered in plants and plant derived products. Intact non-covalent polyphenol-peptide complexes were observed by ESI-MS using different experimental conditions. Utilizing mild ESI interface conditions allowed the detection of 1 : 1 polyphenol-peptide complexes in all tested solutions and 2 : 1 complexes for the dimers and galloylated polyphenols (flavanols). These results show that there is a preferential interaction between polymerized and/or galloylated polyphenols and peptide compared with that between monomeric polyphenols and peptides. Thus, ESI-MS shows potential for the study of small polyphenolic molecule-peptide interactions and determination of stoichiometry.

Topics: Amino Acid Sequence; Catechin; Dimerization; Food Analysis; Macromolecular Substances; Molecular Sequence Data; Peptide Fragments; Peptides; Proline-Rich Protein Domains; Solutions; Spectrometry, Mass, Electrospray Ionization

In the present study, we investigated the free radical scavenging effects of green tea extract and green tea tannin mixture and its components using a nitric oxide (NO) and superoxide (O(2)(-)) generating system in vitro. Green tea extract showed direct scavenging activity against NO and O(2)(-) and green tea tannin mixture, at the same concentration, showed high scavenging activity. Comparison of the activities of seven pure compounds isolated from green tea tannin mixture showed that (-)-epigallocatechin 3-O-gallate (EGCg), (-)-gallocatechin 3-O-gallate (GCg) and (-)-epicatechin 3-O-gallate (ECg) had higher scavenging activities than (-)-epigallocatechin (EGC), (+)-gallocatechin (GC), (-)-epicatechin (EC) and (+)-catechin (C), showing the importance of the structure of flavan-3-ol linked to gallic acid for this activity. Among the gallate-free tannins, EGC and GC were more effective O(2)(-) scavengers than EC and C, indicating the O-trihydroxy structure in the B ring is an important determinant of such activity. However, this structure did not affect the NO scavenging activity. These findings confirm that green tea tannin has excellent antioxidant properties, which may be involved in the beneficial effect of this compound.

Topics: Catechin; Dose-Response Relationship, Drug; Flavonoids; Free Radical Scavengers; In Vitro Techniques; Nitric Oxide; Oxidants; Oxidation-Reduction; Structure-Activity Relationship; Superoxides; Tannins; Tea

Protein oxidation and glycation are posttranslational modifications that are implicated in the pathological development of many age-related disease processes. This study investigated the effects of green tea extract, and a green tea tannin mixture and its components, on protein damage induced by 2,2'-azobis(2-amidinopropane) dihydrochloride (a free radical generator) and glucose in in vitro assay systems. We found that green tea extract can effectively protect against protein damage, and showed that its action is mainly due to tannin. In addition, it was shown that the chemical structures of tannin components are also involved in this activity, suggesting that the presence of the gallate group at the 3 position plays the most important role in the protective activity against protein oxidation and glycation, and that there is also a contribution by the hydroxyl group at the 5' position in the B ring and the sterical structure. These findings demonstrate the mechanisms of the usefulness of green tea in protein oxidation- and glycation-associated diseases.

Topics: Amidines; Antioxidants; Catechin; Free Radicals; Glucose; Glycation End Products, Advanced; Glycosylation; Hydrolyzable Tannins; Oxidants; Oxidation-Reduction; Plant Extracts; Proteins; Tea

Three prolyl endopeptidase (PEP) inhibitors were isolated from the methanolic extract of green tea leaves. They were identified as (-)-epigallocatechin gallate, (-)-epicatechin gallate, and (+)-gallocatechin gallate with the IC50 values of 1.42 x 10(-4) mM, 1.02 x 10(-2) mM, and 1.09 x 10(-4) mM, respectively. They were non-competitive with a substrate in Dixon plots and did not show any significant effects against other serine proteases such as elastase, trypsin, and chymotrypsin, suggesting that they were relatively specific inhibitors against PEP. The isolated compounds are expected to be useful for preventing and curing of Alzheimer's disease.

Topics: Camellia sinensis; Catechin; Kinetics; Magnetic Resonance Spectroscopy; Plant Extracts; Prolyl Oligopeptidases; Protease Inhibitors; Serine Endopeptidases; Spectrometry, Mass, Fast Atom Bombardment; Spectrophotometry, Ultraviolet

Investigators have shown that green tea may decrease the risk of cancer. It is widely accepted that the main active component of green tea is epigallocatechin-3-gallate (EGCG). In this study, we examined the effect of green tea on breast cancer growth and endothelial cells in in vitro assays and in animal models. Furthermore, we compared the potency of the different catechin components of green tea extract (GTE), including EGCG. Our data showed that mixed GTE and its individual catechin components were effective in inhibiting breast cancer and endothelial cell proliferation. In mouse experiments, GTE suppressed xenograft size and decreased the tumor vessel density. Our results demonstrated the value of all catechins and argued for the use of a mixed GTE as a botanical dietary supplement, rather than purified EGCG, in future clinical trials.

Topics: Animals; Breast Neoplasms; Catechin; Cell Division; Cells, Cultured; Endothelium, Vascular; Humans; Mice; Mice, SCID; Plant Extracts; Tea; Transplantation, Heterologous; Umbilical Veins

The title determination was conducted by HPLC with electrochemical detection using an ODS column and a mobile phase of acetonitrile: 0.1 M phosphate buffer (pH 2.5) (15:85, v/v). The eight catechins, gallocatechin (GC), epigallocatechin (EGC), catechin (C), epicatechin (EC), epigallocatechin gallate (EGCg), gallocatechin gallate (GCg), epicatechin gallate (ECg), and catechin gallate (Cg), were detected at 0.6 V vs Ag/AgCl. Good linear relationships between current and amount were noted for 0.5-250 pmol of each catechin, with a correlation coefficient of 0.999 in each case. The detection limit for any one was 0.5 pmol (signal to noise ratio, S/N = 3). After the ingestion of 340 ml canned green tea, GC, EGC, C, and EC, mostly in conjugated form, were determined in urine samples. Conjugated catechins were hydrolyzed by enzymes using sulfatase and beta-glucuronidase. The time courses of the above four catechins showed a maxima at 1-3 h after tea ingestion. (+), (-)-EC and (+), (-)-C were present in canned tea.

Topics: Acetonitriles; Catechin; Chromatography, High Pressure Liquid; Electrochemistry; Flavonoids; Humans; Hydrogen-Ion Concentration; Kinetics; Models, Chemical; Phosphates; Tea; Time Factors