Radical scavenging activities of niacin-related compounds

We investigated whether niacin-related compounds had radical-scavenging activity by electron spin resonance methods. Many compounds, but not trigonelline, had radical-scavenging activity against hydroxyl radicals. However, for the nitric oxide radical and 1,1-diphenyl-2-picrylhydrazyl radical, only nicotinic acid hydrazide and isonicotinic acid hydrazide had scavenging activities. These results suggest that the moiety of hydrazide might have an important role in scavenging abilities of various radicals.     

ESR study on the structure-antioxidant activity relationship of tea catechins and their epimers

The purpose of this study is to examine the relationship between the free radical scavenging activities and the chemical structures of tea catechins ((-)-epigallocatechin gallate (EGCG), (-)-epigallocatechin (EGC) and (-)-epicatechin (EC)) and their corresponding epimers ((-)-gallocatechin gallate (GCG), (-)-gallocatechin (GC) and (+)-catechin ((+)-C)). With electron spin resonance (ESR) we investigated their scavenging effects on superoxide anions (O-.2) generated in the irradiated riboflavin system, singlet oxygen(1O2) generated in the photoradiation-hemoporphyrin system, the free radicals generated from 2,2'-azobis(2-amidinopropane)hydrochloride (AAPH) and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical. The results showed that the scavenging effects of galloylated catechins (EGCG and GCG) on the four free radicals were stronger than those of nongalloylated catechins (EGC, GC, EC, (+)-C), and the scavenging effects of EGC and GC were stronger than those of EC and (+)-C. Thus, it is suggested that the presence of the gallate group at the 3 position plays the most important role in their free radical-scavenging abilities and an additional insertion of the hydroxyl group at the 5' position in the B ring also contributes to their scavenging activities. Moreover, the corresponding phenoxyl radicals formed after the reaction with O-.2 were trapped by DMPO and the ESR spectra of DMPO/phenoxyl radical adducts were observed (aN=15.6 G and aHbeta=21.5 G). No significant differences were found between the scavenging effects of the catechins and their epimers when their concentrations were high. However, significant differences were observed at relatively low concentrations, and the lower their concentrations, the higher the differences. The scavenging abilities of GCG, GC and (+)-C were stronger than those of their corresponding epimers (EGCG, EGC and EC). The differences between their sterical structures played a more important role in their abilities to scavenge large free radicals, such as the free radicals generated from AAPH and the DPPH radical, than to scavenge small free radicals, such as O-.2 and 1O2, especially in the case with EGCG and GCG with more bulky steric hindrance.     

Heat-epimerized tea catechins have the same cholesterol-lowering activity as green tea catechins in cholesterol-fed rats

Tea catechins are known to be epimerized by heat treatment. The effect of heat-epimerized tea catechins on serum cholesterol concentration was compared with that of green tea catechins. Our observations strongly suggest that both tea catechins and heat-epimerized tea catechins lower serum cholesterol concentration by inhibiting cholesterol absorption in the intestine. There was no differential effect between the two catechin preparations.     

Apoptosis-inducing activity of polyphenol compounds derived from tea catechins in human histiolytic lymphoma U937 cells

Polyphenolic compounds derived from tea catechins were examined for apoptosis-inducing activity in human histiolytic lymphoma U937 cells. (-)-Epigallocatechin gallate, theasinensin D, compound OH-5, theaflavin, and theaflavin digallate induced apoptosis as evidenced by DNA ladder formation, its inhibition by a caspase inhibitor, and chromatin condensation. Theasinensin D was the most potent inducer and the data suggest the importance of the number and three dimensional localization of their phenolic groups in this activity. These apoptosis-inducible compounds may be useful as a cancer chemopreventive and chemotherapeutic agent.     

Antibacterial and anti-hemolysin activities of tea catechins and their structural relatives

Among catechins tested, (-)epigallocatechin (EGC), (-)epicatechin gallate (ECg), (-) epigallocatechin gallate (EGCg) inhibited the growth of Staphylococcus aureus, Vibrio cholerae O1 classical Inaba 569B and El Tor Inaba V86. S. aureus was more sensitive than V. cholerae O1 to these compounds. EGCg showed also a bactericidal activity against V. cholerae O1 569B. Pyrogallol showed a stronger antibacterial activity against S. aureus and V. cholerae O1 than tannic and gallic acid. Rutin or caffein had no effect on them. ECg and EGCg showed the most potent anti-hemolysin activity against S. aureus alpha-toxin, Vibrio parahaemolyticus thermostable direct hemolysin (Vp-TDH) and cholera hemolysin. Among catechin relatives, only tannic acid had a potent anti-hemolysin activity against alpha-toxin. These results suggest that the catechol and pyrogallol groups are responsible for the antibacterial and bactericidal activities, while the conformation of catechins might play an important role in the anti-hemolysin activity.     

Inhibition of pectin methyl esterase activity by green tea catechins

Pectin methyl esterases (PMEs) and their endogenous inhibitors are involved in the regulation of many processes in plant physiology, ranging from tissue growth and fruit ripening to parasitic plant haustorial formation and host invasion. Thus, control of PME activity is critical for enhancing our understanding of plant physiological processes and regulation. Here, we report on the identification of epigallocatechin gallate (EGCG), a green tea component, as a natural inhibitor for pectin methyl esterases. In a gel assay for PME activity, EGCG blocked esterase activity of pure PME as well as PME extracts from citrus and from parasitic plants. Fluorometric tests were used to determine the IC50 for a synthetic substrate. Molecular docking analysis of PME and EGCG suggests close interaction of EGCG with the catalytic cleft of PME. Inhibition of PME by the green tea compound, EGCG, provides the means to study the diverse roles of PMEs in cell wall metabolism and plant development. In addition, this study introduces the use of EGCG as natural product to be used in the food industry and agriculture.     

Radical scavenging activity and oxidative modification of citrus dehydrin.

Dehydrins are ubiquitous proteins produced by plants in response to water stress. Their functions, however, are not fully understood. The overexpression of Citrus unshiu Marcov. dehydrin (CuCOR19) enhanced cold tolerance in transgenic plants by reducing lipid peroxidation promoted by cold stress, suggesting that the CuCOR19 protein directly scavenges radicals. In this paper, we report the radical scavenging activity and oxidative modification of CuCOR19. The hydroxyl radical generated by the Fe2+/H2O2 system and peroxyl radical generated from 2, 2'-azobis (2-amidinopropane) (AAPH) were scavenged by CuCOR19, but hydrogen peroxide and superoxide were not. The scavenging activity for the hydroxyl radical and peroxyl radical of CuCOR19 was more potent than that of mannitol, and approximately equal to that of serum albumin, which is known as an antioxidative protein in mammals. CuCOR19 was degraded by the hydroxyl radical and peroxyl radical in a time- and dose-dependent manner. Mannitol and thiourea inhibited the degradation. Analysis of the amino acid composition of CuCOR19 indicated that glycine, histidine, and lysine, which are major residues in many dehydrins, were targeted by the hydroxyl radical. These results suggest that CuCOR19 is a radical scavenging protein, and may reduce oxidative damage induced by water stress in plants.     

Radical scavenging activity of ribonuclease inhibitor from cow placenta

Cow placenta ribonuclease inhibitor (CPRI) has been purified 5062-fold by affinity chromatography, the product being homogeneous by sodium dodecyl sulfate-gel electrophoresis. The chemiluminescence technique was used to determine the radical scavenging activities of CPRI toward different reactive oxygen species (ROS) including superoxide anion (O2-*), hydroxyl radical (OH*), lipid-derived radicals (R*), and singlet oxygen (1O2). CPRI could effectively scavenge O2-*, OH*, R*, and 1O2 at EC50 of 0.12, 0.008, 0.009, and 0.006 mg/ml, respectively. In addition, the radical scavenging activities of CPRI were higher than those of tea polyphenols, indicating that CPRI is a powerful antioxidant.     

中华补血草多酚提取物对自由基的清除能力

采用超声波辅助浸提—大孔树脂吸附法从中华补血草[ Limonium sinense (Girard) Kuntze]根、根茎、叶和花中获得多酚提取物,并对它们的得率和组成成分进行了初步分析;在此基础上,比较研究了不同部位多酚提取物对DPPH·、·OH和O2的清除能力.实验数据表明:中华补血草根、根茎、叶和花多酚提取物的得率分别为12.42％、5.98％、5.27％和3.98％,差异明显;多酚提取物中总酚、原花色素、黄烷醇和总黄酮含量变化幅度较大,分别为51.87％ ～61.60％、5.62％～39.47％、3.69％～12.46％和2.53％ ～35.97％;其中,总酚、原花色素和黄烷醇含量均以根多酚提取物最高,总黄酮含量以花多酚提取物最高,均与其他部位多酚提取物有显著差异.随质量浓度提高,4个部位多酚提取物对3种自由基的清除率总体上逐渐增大;其中根多酚提取物对DPPH·的清除作用最强,半数清除质量浓度(ρSC50)为38.52 μg·L-1,显著低于阳性对照芦丁和BHT(ρSC50分别为67.40和74.25 μg·L-1);花多酚提取物对·OH和O2的清除能力最强,ρSC50分别为53.51和74.00 μg·L-1,均低于芦丁;总体上,4个部位多酚提取物对DPPH·的清除能力由强至弱依次为根、根茎、叶、花,对·OH和O2的清除能力由强至弱依次为花、根、叶、根茎.结果显示:中华补血草不同部位多酚提取物均具有较强的自由基清除能力,其中,根和花多酚提取物中高含量的原花色素和黄酮类成分分别与其自由基清除能力密切相关.     

Radical scavenging activity of phenylpropanoid glycosides in Caryopteris incana.

In our screening program for antioxidants from traditional drugs and foodstuffs, one new phenylpropanoid glycoside, incanoside, was isolated together with four known phenylpropanoid glycosides, verbascoside, isoverbascoside, phlinoside A, and 6-O-caffeoyl-beta-D-glucose from the whole plant of Caryopteris incana (Thunb.) Miq. On the basis of chemical evidence and spectral analysis data, the structure of incanoside was determined to be 1-O-(3,4-dihydroxyphenyl)ethyl-O-beta-D-glucopyranosyl (1-->2)-alpha-L-rhamnopyranosyl(1-->3)-6-O-caffeoyl-beta-D- glucopyranoside. The four phenylpropanoid glycosides exhibited potent radical scavenging activity against DPPH, hydroxyl (.OH), and superoxide anion (O2-.) radicals.     

Plasma and lipoprotein levels of tea catechins following repeated tea consumption

Epidemiological studies suggest that antioxidant flavonoids in tea may reduce the risk of cardiovascular disease, possibly via protection of low-density lipoproteins (LDL) against oxidation. However, the extent of absorption of tea flavonoids and their accumulation in LDL during regular consumption of tea is not clear. Therefore we investigated plasma and lipoprotein levels of catechins during tea consumption and the impact on LDL oxidizability ex vivo. Eighteen healthy adults consumed, in an incomplete balanced cross-over design, green tea, black tea, black tea with milk or water, one cup every 2 hr (eight cups/day) for three days. Blood samples were obtained in the mornings and evenings of each day. Plasma total catechin concentration was determined in all blood samples, and the distribution of catechins among lipoproteins was determined at the end of the third day (t = 60 hr). The resistance of LDL to copper-induced oxidation ex vivo was assessed before tea consumption and at t = 60 hr. Repeated tea consumption during the day rapidly increased plasma total catechin levels whereas they declined overnight when no tea was consumed. There was a gradual increase in plasma levels in the mornings (respectively, 0.08 microM vs. 0.20 microM on first and last day of black tea consumption) and evenings (respectively, 0.29 microM vs. 0.34 microM on first and last day of black tea consumption). Green tea catechins were mainly found in the protein-rich fraction of plasma (60%) and in high-density lipoproteins (23%). Although present in LDL, the concentration of catechins in LDL was not sufficient to enhance the resistance of LDL to oxidation ex vivo. Addition of milk to black tea did not affect any of the parameters measured. In conclusion, the present study shows that catechin levels in blood rapidly increase upon repeated tea consumption. The accumulation of catechins in LDL particles is not sufficient to improve the intrinsic resistance of LDL to oxidation ex vivo.     

Green tea catechins can bind and modify ERp57/PDIA3 activity

Background

Green tea is a rich source of polyphenols, mainly catechins (flavanols), which significantly contribute to the beneficial health effects of green tea in the prevention and treatment of various diseases. In this study the effects of four green tea catechins on protein ERp57, also known as protein disulfide isomerase isoform A3 (PDIA3), have been investigated in an in vitro model.

Methods

The interaction of catechins with ERp57 was explored by fluorescence quenching and surface plasmon resonance techniques and their effect on ERp57 activities was investigated.

Results

A higher affinity was observed for galloylated cathechins, which bind close to the thioredoxin-like redox-sensitive active sites of the protein, with a preference for the oxidized form. The effects of these catechins on ERp57 properties were also investigated and a moderate inhibition of the reductase activity of ERp57 was observed as well as a strong inhibition of ERp57 DNA binding activity.

Conclusions

Considering the high affinity of galloylated catechins for ERp57 and their capability to inhibit ERp57 binding to other macromolecular ligands, some effects of catechins interaction with this protein on eukaryotic cells may be expected.

General significance

This study provides information to better understand the molecular mechanisms underlying the biological activities of catechins and to design new polyphenol-based ERp57-specific inhibitors.     

Radical scavenging activity of protein from tentacles of jellyfish Rhopilema esculentum

In this study, radical scavenging activity of protein from tentacles of jellyfish Rhopilema esculentum (R. esculentum) was assayed including superoxide anion radical and hydroxyl radical scavenging. The protein samples showed strong scavenging activity on superoxide anion radical and values EC50 of full protein (FP), first fraction (FF), second fraction (SF), and 30% (NH4)2 SO4 precipitate (Fr-1) were 2.65, 7.28, 1.10, and 22.51 microg/mL, respectively, while values EC50 of BHA, BHT, and alpha-tocopherol were 31, 61, and 88 microg/mL, respectively. Also, the protein samples had strong scavenging effect on hydroxyl radical and the values EC50 of FP, FF, SF, Fr-1, and Fr-2 were 48.91, 27.72, 1.82, 16.36, and 160.93 microg/mL, but values EC50 of Vc and mannitol were 1907 and 4536 microg/mL, respectively. Of the five protein samples, SF had the strongest radical scavenging activity and may have a use as a possible supplement in the food and pharmaceutical industries. The radical scavenging activity was stable at high temperature so that R. esculentum may be used as a kind of natural functional food.     

Radical scavenging ability of some compounds isolated from Piper cubeba towards free radicals

The purpose of this study was to identify the antioxidant activity of 16 compounds isolated from Piper cubeba (CNCs) through the extent of their capacities to scavenge free radicals, hydroxyl radical (HO^?), superoxide anion radical () and 2,2‐diphenyl‐1‐picrylhydrazyl radical (DPPH^?), in different systems. Electron paramagnetic resonance (EPR) and 5,5‐dimethyl‐1‐pyrroline‐N‐oxide, DMPO, as the spin trap, and chemiluminescence techniques were applied. Using the Fenton‐like reaction [Fe(II) + H₂O₂], CNCs were found to inhibit DMPO? OH radical formation ranging from 5 to 57% at 1.25 mmol L^?1 concentration. The examined CNCs also showed a high DPPH antiradical activity (ranging from 15 to 99% at 5 mmol L^?1 concentration). Furthermore, the results indicated that seven of the 16 tested compounds may catalyse the conversion of superoxide radicals generated in the potassium superoxide/18‐crown‐6 ether system, thus showing superoxide dismutase‐like activity. The data obtained suggest that radical scavenging properties of CNCs might have potential application in many plant medicines. Copyright © 2010 John Wiley & Sons, Ltd.     

Radical scavenging properties of genistein

The reactivity of genistein toward reactive radical species has been investigated by means of pulse radiolysis. The values of rate constants, respectively 2.3 x 10(10) M(-1)s(-1) and 1.3 x 10(10) M(-1)s(-1) for the reaction with hydroxyl radical at pH 8.3 and 3.0, are close to diffusion limit indicating that genistein is a potent hydroxyl radical scavenger. The reactivity of genistein towards one-electron oxidants has also been investigated. The rate constants k = 4.6 x 10(9) M(-1)s(-1) (pH 8.3) and 6.7 x 10(8) M(-1)s(-1) (pH 7.6) have been determined for the reaction of genistein with *N3 and Br2*- radicals, respectively. For both oxidants the rate constants at pH 3 does not exceed 10(8) M(-1)s(-1). The differences in reactivity of genistein towards the oxidants at different acidity of the solution have been assumed to arise from the acid-base equilibria of genistein. The dissociation constants for genistein (pKa: 7.2, 10.0, and 13.1) have been evaluated spectroscopically. The influence of acid-base equilibria on bond dissociation energy and ionization potential for genistein has also been investigated by means of DFT calculations. It has been concluded on the basis of these calculations that monoanionic form of genistein existing at physiological pH is more powerful radical scavenger than the neutral molecule.     

Radical scavenging properties of polyamines

Di- and polyamines are effective scavengers of free radicals generated in a number of chemical and in vitro enzyme systems. Free radical production was quantified spectrophotometrically using nitroblue tetrazolium and cytochrome c or by electron spin resonance. Levels of superoxide radical formed either enzymatically with xanthine oxidase or chemically from riboflavin or pyrogallol were significantly inhibited by spermine, spermidine, putrescine and cadaverine at 10 and 50 mM. The more reactive hydroxyl radical generated by the Fenton reaction was also effectively scavenged by di- and polyamines. In addition, the production of superoxide radical by senescing microsomal membranes was inhibited by di- and polyamines, as was the superoxide-dependent conversion of 1- aminocyclopropane-1-carboxylic acid (ACC) to ethylene. The efficacy of polyamine-scavenging appears to be correlated with the extent of amination suggesting the involvement of amino groups. It is also apparent that some of the physiological effects of polyamines, in particular their propensity to inhibit lipid peroxidation and retard senescence, may be attributable to their radical-scavenging capability.     

Polyphenolic compounds from Geranium pratense and their free radical scavenging activities

Two new polyphenolic compounds, myricetin 3-O-(2"-O-galloyl)-beta-D-glucopyranoside and (-)-6-chloroepicatechin, were isolated from the aerial parts of Geranium pratense subsp. finitimum (Woronow) Knuth, along with three known polyphenolic compounds [quercetin 3-O-(2"-O-galloyl)-beta-D-glucopyranoside, quercetin 3-O-(2"-O-galloyl)-beta-D-galactopyranoside, methyl gallate] and tryptophan. Quercetin 3-O-beta-D-glucopyranoside, quercetin 3-O-beta-D-galactopyranoside, quercetin 3-O-(2"-O-galloyl)-beta-D-glucopyranoside and quercetin 3-O-(2"-O-galloyl)-beta-D-galactopyranoside were found to be effective against free radical induced impairment of endothelium-dependent relaxation in isolated rat aorta.