Redifferentiation of human hepatoma cells induced by green tea polyphenols

A novel approach for the treatment of cancer is the differentiation therapy in which cancer cells are induced to attain a mature phenotype when exposed to differentiation inducers. To examine the effects of polyphenols extracted from green tea, i.e. ( – )-epicatechin (EC), ( – )-epigallocatechin (EGC), ( – )-epicatechin gallate (ECG) and ( – )-epigallocatechin gallate (EGCG), on the proliferation and redifferentiation of human hepatoma cell line SMMC-7721, we measured the changes of cell growth, cell surface charge and cell morphography after treament with green tea polyphenols. It was found that the growth curve of treated cells was decreased remarkably, cell surface charge of treated cells was decreased and the microvilli on the surface of treated cells were reduced obviously. It confirmed that green tea polyphenols could reverse malignant phenotypic characteristics and induced redifferentiation of SMMC-7721 cells. The ability of green tea polyphenols to inhibit reactive oxygen species (ROS)-mediated oxidative damage of DNA was also assessedin vitro by measuring the conversion of supercoiled pBR322 plasmid DNA to the open circular and linear forms. It was found that green tea polyphenols could significantly inhibit the oxidative damage of DNA induced by a water-soluble azo initiator 2,2-azobis(2-amidinopropane hydrochloride) (AAPH). However, they could promote the oxidative damage of DNA induced by H₂O₂ and Fe²⁺ at high concentrations. The relationship between the anti-cancer activity and antioxidation-prooxidation activity of green tea polyphenols is discussed.     

红茶、绿茶和乌龙茶多糖及多酚对DNA的保护作用研究

用溴化乙锭(EB)作为荧光探针研究了各种茶叶多糖、多酚类化合物对DNA的保护作用.测定了龙井、普耳茶、乌龙茶等8种茶叶中多糖和多酚类提取物存在下DNA与EB混合液的荧光积分强度,并把不同茶叶与DNA相互作用的常数定义为结合常数D,根据D的大小讨论了多糖及多酚类化合物的存在DNA保护作用的影响.结果表明,8种茶叶与DNA...     

Antioxidant activity determination in Sencha and Gun Powder green tea extracts with the application of voltammetry and UV-VIS spectrophotometry

This article presents the electrochemical analysis of the oxidation of rutin and green tea extract solutions and the antioxidative properties of these substances. Flavonoids and other polyphenols contained in green tea have antioxidative properties due to the presence of various numbers of hydroxyl groups in different arrangements. The investigation of the oxidation of green tea extracts was performed to identify the most effective antioxidant. The cyclic and pulse voltammograms show that the extract of Gun Powder (GP) green tea contained rutin and other polyphenols, while the extract of Sencha (S) tea contained other antioxidants that are oxidised at a more positive potential. The GP extract showed slightly better antioxidative properties than did the S extract. The UV-VIS spectra show that, in addition to flavonoids extracts contain chlorophyll. The results obtained demonstrate that the tested tea extracts show very good antioxidative properties and therefore may be considered as potential stabilising agents that are able to reduce the rate of undesirable oxidation processes.     

Simultaneous determination of total polyphenols and caffeine contents of green tea by near-infrared reflectance spectroscopy

This paper indicates the possibility to use near infrared (NIR) spectroscopy as a rapid method to predict quantitatively the content of caffeine and total polyphenols in green tea. A partial least squares (PLS) algorithm is used to perform the calibration. To decide upon the number of PLS factors included in the PLS model, the model is chosen according to the lowest root mean square error of cross-validation (RMSECV) in training. The correlation coefficient R between the NIR predicted and the reference results for the test set is used as an evaluation parameter for the models. The result showed that the correlation coefficients of the prediction models were R = 0.9688 for the caffeine and R = 0.9299 for total polyphenols. The study demonstrates that NIR spectroscopy technology with multivariate calibration analysis can be successfully applied as a rapid method to determine the valid ingredients of tea to control industrial processes.     

绿茶多酚对自由基诱导的红细胞膜过氧化的抑制作用

采用水溶性偶氮引发剂2,2'-偶氮二(2-脒基丙烷)二盐酸盐(AAPH)在37引发入血红细胞膜的过氧化,通过测定氧气吸收及维生素E的消耗研究了过氧化过程的动力学,并对从绿茶中提取的主要多酚类化合物的抗氧化活性做了定量研究。使用的绿茶多酚有:(-)-表儿茶素(EC),(-)-表儿茶素(EGC),(-)-表儿茶素酸酯(ECG)和(-)-表儿茶素培酸酯(EGCG)。结果表明,这些绿茶多酚能够显著缩短过氧化反应的动力学链长,有效地抑制红细胞膜的过氧化。抗氧化活性顺序为:EC〉GCG〉EGCG〉EGC。     

Antioxidant and antimicrobial activities of green tea extract loaded into nanostructured lipid carriers

In this paper, the preparation and characterization of some novel nanostructured lipid carriers for drug delivery are reported. They are obtained by mixing two solid lipids, cetyl palmitate and glyceryl stearate, with three types of vegetable oils: grape seed oil, St. John's wort oil (Hypericum perforatum oil) and sea buckthorn oil. In order to increase their antioxidant and antimicrobial properties, they are co-loaded with green tea extract by using a modified high shear homogenization technique. Size distribution and polydispersity index of the developed nanostructured lipid carriers determined by the dynamic light scattering, and corroborated with the results obtained by the transmission electron microscopy analysis, confirmed that the structures obtained are at nanoscales. The crystallinity behavior of the prepared nanostructured lipid carriers has been studied by differential scanning calorimetry; zeta potential measurements show that all loaded nanostructures present excellent physical stability. Their antioxidant and antimicrobial properties evaluated by an appropriate in vitro analysis using the chemiluminescence method, and the diffusion disc method, respectively, show that green tea extract could be utilized as a valuable natural source of antioxidant and antimicrobial agent. These new nano-formulations proved to have significant potential for nutritional and pharmaceutical applications.     

Toxicologic damage of gas phase cigarette smoke on cells and the protective effect of green tea polyphenols

A mouse fibroblast cell line, Chinese hamster lung V₇₉ cell, was used to assess the toxicological effect of gas phase cigarette smoke (GPCS) on cells and the protective effect of green tea polyphenols (GTP). Exposure of the cultured V₇₉ cells to GPCS decreased cell viability and caused lipid peroxidation of cell membrane as measured by thiobarbituric acid reaction. Electron spin resonance (ESR) spin trapping and spin labeling studies indicated that GPCS exposure could increase the fluidity in the polar surface of cell membrane, but the membrane fluidity in the hydrophobic region was not affected. The conformation of sulfhydryl binding sites on membrane was changed. GTP itself had no effect on the polar surface of membrane nor changed the conformation of sulfhydryl binding sites on membrane proteins in the concentration used in this experiment. Incubation of V₇₉ cells with GTP aqueous solution before GPCS treatment could decrease cell death and lipid peroxidation and inhibit the changes of the biophysical properties of cell membrane induced by GPCS in a dose-dependent manner. Also GTP could scavenge the free radicals generated by GPCS. These results indicate that GPCS can induce lipid peroxidation and affect the biophysical properties of cell membrane, which may account for the toxicity of GPCS, and that GTP can prevent these changes possibly by scavenging the free radicals in GPCS and inhibiting the membrane lipid peroxidation.     

Pressurized liquid extraction of mate tea leaves

The objective of this work is to investigate the influence of process parameters on the pressurized liquid extraction (PLE) of Ilex paraguariensis leaves. A factorial 2⁶⁻² experimental design was employed using responses as the extraction yield and the chromatographic profile of the extracts. The extraction time, polarity of solvent, amount of sample, numbers of PLE cycles, flushing volume and extraction temperature were selected as independent variables (factors). Results obtained indicated that the solvent polarity was the most significant variable in the study, while the amount of sample and extraction temperature also showed significant effect. The other variables did not present significant influence in the yield of extraction. GC/MS analysis of the extract enabled the identification of saturated hydrocarbons, fatty acids, fatty acid methyl esters, phytosterols and theobromine in the extracts. Quantitative analysis of four compounds presented in the extracts (caffeine, phytol, vitamin E and squalene) was performed by the GC/MS in the SIM mode.     

Antioxidant potential of polyphenols and tannins from burs of Castanea mollissima Blume

Spiny burs of Castanea mollissima Blume (Chinese chestnut) are usually discarded as industrial waste during post-harvesting processing. The objective of this study was to establish an extraction and isolation procedure for tannins from chestnut burs, and to assess their potential antioxidant activity. Aqueous ethanol solution was used as extraction solvent, and HPD 100 macroporous resin column was applied for isolation. The influence of solvent concentration in the extraction and elution process on extraction yield, tannins and polyphenols content, as well as antioxidant potential, including DPPH and ABTS radical scavenging ability, reducing power ability and cellular antioxidant ability were assessed. In both the extraction and isolation process, 50% aqueous ethanol led to superior total tannins and polyphenols content as well as significantly higher antioxidant activity. In addition, the antioxidant activity and the total tannins content in extracts and fractions had a positive linear correlation, and the predominant components responsible for antioxidant activities were characterized as hydrolysable tannins. To the best of our knowledge, this is the first report on the enrichment of tannins from burs of C. mollissim using macroporous resin chromatography, and to assess the cellular antioxidant activity of them.     

Effect of tea polyphenols on glucan synthesis by glucosyltransferase from Streptococcus mutans

In the course of our studies on the development of anti-plaque agents for prevention of dental caries, we investigated effects of some of tea preparations and their individual components on the glucan synthesis catalyzed by glucosyltransferase (GTF) from Streptococcus mutans. Extracts of green tea and black tea, and polyphenol mixtures showed appreciable inhibition in the synthesis of insoluble glucan. Among the components isolated from tea infusions, theaflavin and its mono- and digallates had potent inhibitory activities at concentrations of 1-10 mM against GTF. (+)-Catechin, (-)-epicatechin and their enantiomers had moderate inhibitory activities at these concentrations, while galloyl esters of (-)-epicatechin, (-)-epigallocatechin and (-)-gallocatechin had increased inhibitory activities.     

Green tea polyphenols: DNA photodamage and photoimmunology.

Green tea is a popular beverage consumed worldwide. The epicatechin derivatives, which are commonly called 'polyphenols', are the active ingredients in green tea and possess antioxidant, anti-inflammatory and anti-carcinogenic properties. Studies conducted by our group on human skin have demonstrated that green tea polyphenols (GTP) prevent ultraviolet (UV)-B-induced cyclobutane pyrimidine dimers (CPD), which are considered to be mediators of UVB-induced immune suppression and skin cancer induction. GTP treated human skin prevented penetration of UV radiation, which was demonstrated by the absence of immunostaining for CPD in the reticular dermis. The topical application of GTP or its most potent chemopreventive constituent (-)-epigallocatechin-3-gallate (EGCG) prior to exposure to UVB protects against UVB-induced local as well as systemic immune suppression in laboratory animals. Additionally, studies have shown that EGCG treatment of mouse skin inhibits UVB-induced infiltration of CD11b+ cells. CD11b is a cell surface marker for activated macrophages and neutrophils, which are associated with induction of UVB-induced suppression of contact hypersensitivity responses. EGCG treatment also results in reduction of the UVB-induced immunoregulatory cytokine interleukin (IL)-10 in skin as well as in draining lymph nodes, and an elevated amount of IL-12 in draining lymph nodes. These in vivo observations suggest that GTPs are photoprotective, and can be used as pharmacological agents for the prevention of solar UVB light-induced skin disorders associated with immune suppression and DNA damage.     

Efficient green extraction of polyphenols from post-harvested agro-industry vegetal sources in Piedmont

A study of the polyphenols content and antioxidant capacity of grapevine waste and hazelnut skins (roasted material) from post-harvest products that originate from Piedmont (Italy) has been carried out and the results herein presented. Ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) were used to achieve process intensification in shorter extraction times, with lower environmental impact and higher selectivity compared to classic maceration. Besides classic solvents, the aqueous β-cyclodextrin solution (1.5%) showed to be an excellent extraction medium for grapevine waste. Total phenolic content (TP) from grapevine waste ranged from 18.23 ± 2.4 to 198 ± 3 mg gallic acid equivalents (GAE)/g dry weight, while total antioxidant capacity (TAC) expressed as EC₅₀ ranged from 0.0902 ± 0.08 mg/mL to 0.0041 ± 0.02 mg/mL. For hazelnut skins, TP ranged from 61.68 ± 0.8 to 200.79 ± 3.0 mg GAE/g dry weight, while TAC ranged from 0.0021 ± 0.0004 to 0.0002 ± 0.0001 mg/mL extract. We have shown that, compared to maceration, the use of UAE and MAE methods can enhance polyphenols recovery and antioxidant capacity.     

Antioxidant activity,lipophilicity and extractability of polyphenols from pig skin – development of analytical methods for skin permeation studies

Permeation of polyphenols through the stratum corneum barrier is a precondition for the protective action of polyphenols against oxidative skin damage. Prior to in vitro skin permeation experiments, we developed a method for the quantification of polyphenols in pig skin, including organic solvent extraction and HPLC analysis. Catechine hydrate, epigallocatechin gallate, trans‐resveratrol, quercetin, rutin and protocatechuic acid were chosen for this study as representatives of phenolics with different lipophilicity and molecular weight. The antioxidative activities of polyphenols as well as their octanol–water partition coefficients at different pH values were determined. Extraction of polyphenols from pig skin was optimized by variation of solvent composition, homogenization intensity and time, as well as partial exclusion of oxygen during extraction. The highest recovery rates could be reached by extraction with the methanol–water mixture (90:10, v/v), containing 0.2 g/L l ‐ascorbic acid, after the cryo‐milling for 4 min. Recoveries of 72% for total phenolics, 96% for quercetin and protocatechuic acid, 90% for rutin and 74% for trans‐resveratrol, were achieved. These extraction parameters will be selected for the polyphenol extraction from pig skin for further in vitro drug permeation studies. Copyright © 2013 John Wiley & Sons, Ltd.     

Investigation of the radioprotective properties of some tea polyphenols

The influence of tea and coffee flavonoids (−)-epicatechin (EC) and (+)-catechin (C) on the effect of γ-irradiation of DNA molecules in solutions was investigated. The concentrations of these and other components in different samples of black and green tea were analyzed by the capillary electrophoresis. It was determined from the experimental data obtained by different methods (circular dichroism and UV spectra, viscosimetry and optical investigations) that C and EC do not form complexes with DNA in solutions. It was shown that C and EC have radioprotective properties, being scavengers of the water radiolysis products.     

茶叶中的茶多酚荧光检测方法

以石墨烯量子点为荧光探针,基于二氧化锰纳米片与石墨烯量子点之间发生有效的荧光共振能量转移,构建了一种茶叶中茶多酚含量的荧光检测新方法。MnO2纳米片使石墨烯量子点的荧光淬灭,而茶多酚能与MnO2发生氧化还原反应,将其还原成Mn^2+,使体系荧光恢复。茶多酚含量与荧光强度增量在20~750μg/mL范围内成线性相关,线性方程为F=1.0574c-114.9,线性相关系数为R=0.9936。将该方法用于茶叶中茶多酚含量的检测,结果与国标法接近。方法有望应用于食品中其它抗氧化物质的实时检测。     

猫须草多酚的提取工艺优化及其抗氧化活性

研究猫须草多酚的提取工艺及其抗氧化活性。通过单因素试验探讨提取剂、乙醇体积分数、料液比、提取温度、提取时间、提取次数等因素对猫须草多酚提取率的影响,在单因素试验的基础上进行正交试验,结果表明,猫须草多酚的最佳提取工艺条件如下:10%乙醇,料液比1∶30,提取温度90℃,提取时间40min,在该条件下猫须草多酚的提取率为11.99mg·g-1。通过测定猫须草多酚清除DPPH自由基、超氧阴离子自由基、亚硝酸根离子的能力及其还原力,对猫须草多酚的抗氧化活性进行评价,结果表明,在一定质量浓度范围内,猫须草多酚清除DPPH自由基的能力及还原力优于Vc,清除超氧阴离子自由基的能力与Vc接近,但清除亚硝酸根离子的能力低于Vc。     

钼酸铵分光光度法测定绿茶中的茶多酚

采用钼酸铵分光光度法测定茶叶中的茶多酚含量。它是基于茶多酚在酸性条件下能与钼酸铵反应生成黄色钼酸酯,此物质在341nm处有最大吸收,用分光光度法测其吸光度,从而间接测定茶多酚含量。方法的线性范围为4×10-6～36×10-5g mL,ε=2.1×105L mol·cm,RSD=1 3%(茶多酚含量8×10-6g mL,n=11)。测定结果与标准法基本一致。     

Immobilised tyrosinase-based biosensor for the detection of tea polyphenols

An amperometric principle-based biosensor containing immobilized enzyme tyrosinase has been used for detection of polyphenols in tea. The immobilized tyrosinase-based biosensor could detect tea polyphenols in the concentration range 10–80 mmol L⁻¹. Immobilization of the enzyme by the crosslinking method gave good stable response to tea polyphenols. The biosensor response reached the steady state within 5 min. The voltage response was found to have a direct linear relationship with the concentration of polyphenols in black tea samples. Enzyme membrane fouling was observed with number of analyses with a single immobilised enzyme membrane. The tyrosinase-based biosensor gave maximum response to tea polyphenols at 30°C. The optimum pH was 7.0. This biosensor system can be applied for analysis of tea polyphenols. Variation in the biosensor response to black tea infusions gave an indication of the different amounts of theaflavins in the samples, which is an important parameter in evaluating tea quality. A comparative study of the quality attributes of a variety of commercially available brands of tea were performed using the biosensor and conventional analytical techniques such as spectrophotometry.     

Optimum conditions for the water extraction of L-theanine from green tea

Theanine is a unique non-protein amino acid found in tea (Camellia sinensis). It contributes to the favourable umami taste of tea and is linked to various beneficial effects in humans. There is an increasing interest in theanine as an important component of tea, as an ingredient for novel functional foods and as a dietary supplement. Therefore, optimal conditions for extracting theanine from tea are required for the accurate quantification of theanine in tea and as an efficient first step for its purification. This study examined the effects of four different extraction conditions on the yield of theanine from green tea using water and applied response surface methodology to further optimise the extraction conditions. The results showed that temperature, extraction time, ratio of water-to-tea and tea particle sizes had significant impacts on the extraction yield of theanine. The optimal conditions for extracting theanine from green tea using water were found to be extraction at 80 °C for 30 min with a water-to-tea ratio of 20:1 mL/g and a tea particle size of 0.5-1 mm.     

Synthesis, characterization and cytotoxicity studies of chitosan-coated tea polyphenols nanoparticles

Chitosan nanoparticles (CS-NPs) were prepared by ionic gelation method using carboxymethyl chitosan and chitosan hydrochloride as carriers of tea polyphenols. The characteristics of chitosan-coated tea polyphenols nanoparticles (CS-TP NPs) were determined by using transmission electron microscopy (TEM) and FT-IR spectroscopy. It was found that the synthesized CS-TP NPs were non-spherical in shape with an average size of 407±50nm. Meanwhile, the drug content and encapsulation rate of the nanoparticles was 8-16% and 44-83%, respectively. These CS-TP NPs also demonstrated sustained release of tea polyphenols in PBS. The antitumor of CS-TP NPs towards HepG2 cancer cells was investigated. The result showed that CS-TP NPs retained significant antitumor activities.