Xavier Vitrac

Comparative Study of Antioxidant Properties and Total Phenolic Content of 30 Plant Extracts of Industrial Interest Using DPPH, ABTS, FRAP, SOD, and ORAC Assays

Aqueous extracts of 30 plants were investigated for their antioxidant properties using DPPH and ABTS radical scavenging capacity assay, oxygen radical absorbance capacity (ORAC) assay, superoxide dismutase (SOD) assay, and ferric reducing antioxidant potential (FRAP) assay. Total phenolic content was also determined by the Folin-Ciocalteu method. Antioxidant properties and total phenolic content differed significantly among selected plants. It was found that oak (Quercus robur), pine (Pinus maritima), and cinnamon (Cinnamomum zeylanicum) aqueous extracts possessed the highest antioxidant capacities in most of the methods used, and thus could be potential rich sources of natural antioxidants. These extracts presented the highest phenolic content (300-400 mg GAE/g). Mate (Ilex paraguariensis) and clove (Eugenia caryophyllus clovis) aqueous extracts also showed strong antioxidant properties and a high phenolic content (about 200 mg GAE/g). A significant relationship between antioxidant capacity and total phenolic content was found, indicating that phenolic compounds are the major contributors to the antioxidant properties of these plants.

Distribution of [14C]-trans-resveratrol, a cancer chemopreventive polyphenol, in mouse tissues after oral administration

Trans-resveratrol, a phenolic compound present in wine, has been reported to be a potential cancer chemopreventive agent. However, although it has numerous biological activities in vitro, there are few data about its bioavailability and tissue distribution in vivo. The objectives of this study were to investigate the absorption and tissue distribution of 14C-trans-resveratrol following oral administration to mice. Male Balb/c mice were given a single oral dose of 14C-trans-resveratrol and were sacrificed at 1.5, 3 or 6 h postdose. The distribution of radioactivity in tissues was evaluated using whole-body autoradiography, quantitative organ-level determination and microautoradiography. In addition, identification of radioactive compounds in kidney and liver was done with high-performance liquid chromatography. Autoradiographic survey of mice sections as well as radioactivity quantification in various organs revealed a preferential fixation of 14C-trans-resveratrol in the organs and biological liquids of absorption and elimination (stomach, liver, kidney, intestine, bile, urine). Moreover, we show that 14C-trans-resveratrol derived radioactivity is able to penetrate the tissues of liver and kidney, a finding supported by microautoradiography. The presence of intact 14C-trans-resveratrol together with glucurono- and/or sulfoconjugates in these tissues was also shown. This study demonstrates that trans-resveratrol is bioavailable following oral administration and remains mostly in intact form. The results also suggest a wide range of target organs for cancer chemoprevention by wine polyphenols in humans.

Direct liquid chromatographic analysis of resveratrol derivatives and flavanonols in wines with absorbance and fluorescence detection

Abstract Wine contains a large number of polyphenols including stilbenes, flavanols and anthocyanins. Of these, stilbenes have been reported to have potential chemopreventive activities. We describe the simultaneous determination of six resveratrol derivatives in wines by liquid chromatography (LC) with fluorescence detection. The levels of pallidol, the symmetrical dimer of resveratrol, are reported for the first time. Quantifications were carried out at optimal wavelengths for each compound during separation. A total of 19 red and 30 white commercial wines from South-Western France were analysed, and the highest stilbene concentrations were found in red wines. In addition, the levels of catechins and two flavanonols recently isolated in red wine are reported.

Integrin αVβ3 contains a receptor site for resveratrol

Resveratrol is a naturally occurring polyphenol, which causes apoptosis in cultured cancer cells. We describe a cell surface resveratrol receptor on the extracellular domain of hetero‐dimeric αVβ3 integrin in MCF‐7 human breast cancer cells. This receptor is linked to induction by resveratrol of extracellular‐regulated kinases 1 and 2 (ERK1/2)‐ and serine‐15‐p53‐dependent phosphorylation leading to apoptosis. The integrin receptor is near the Arg‐Gly‐Asp (RGD) recognition site on the integrin; an integrin‐binding RGD peptide inhibits induction by resveratrol of ERK1/2‐and p53‐dependent apoptosis. Antibody (Ab) to integrin αVβ3, but not to αVβ5, inhibits activation by resveratrol of ERK1/2 and p53 and consequent apoptosis in estrogen receptor‐α (ERα) positive MCF‐7, and ERα‐negative MDA‐MB231 cells. Resveratrol is displaced from the purified integrin by an RGD, but not RGE, peptide, and by αVβ3 integrin‐specific Ab. Resveratrol action is blocked by siRNAβ3, but not by siRNAαV. [14C]‐Resveratrol binds to commercially purified integrin αVβ3 and to αVβ3 prepared from MCF‐7 cells;binding of [14C]‐resveratrol to the β3, but not to the αV monomer, is displaced by unlabeled resveratrol. In conclusion, binding of resveratrol to integrin αVβ3, principally to the β3 monomer, is essential for transduction of the stilbene signal into p53‐dependent apoptosis of breast cancer cells.—Lin, H.‐Y., Lansing, L., Merillon, J.‐M., Davis, F. B., Tang, H.‐Y., Shih, A., Vitrac, X., Krisa, S., Keating, T., Cao, H. J., Bergh, J., Quackenbush, S., Davis, P. J. Integrin αVβ3 contains a receptor site for resveratrol. FASEB J. 20, E1133–E1138 (2006)

Phenolic composition and antioxidant properties of poplar bud (Populus nigra) extract: individual antioxidant contribution of phenolics and transcriptional effect on skin aging.

The Populus species possess great potential for therapeutical applications, especially for their known anti-inflammatory properties. The antioxidant properties of propolis, a hive product collected by honey bees mainly from poplar bud exudates, suggest that poplar buds also possess antioxidant properties. Here is reported the characterization of the antioxidant properties of an aqueous poplar bud (Populus nigra) extract. It presented a high total phenolic content, and moderate antioxidant properties as determined by ORAC assay. The main phenolic compounds identified were phenolic acids and flavonoid aglycons. These phenolic compounds were analyzed by ORAC assay for their individual antioxidant activity, in order to determine the major contributors to the total antioxidant activity of the extract. Thanks to their high antioxidant activity, caffeic and p-coumaric acids were identified as the major antioxidant components. Representing only 3.5% of its dry weight, these compounds represented together about 50% of the total antioxidant activity of the extract. The antioxidant properties of poplar bud extract and the phenolic compounds identified were also analyzed by cellular antioxidant activity assay (CAA), which was weakly correlated with ORAC assay. The transcriptional effect of poplar bud extract on skin aging was evaluated in vitro on a replicative senescence model of normal human dermal fibroblasts, using a customized DNA macroarray specifically designed to investigate skin aging markers. Among the detected genes, poplar bud extract significantly regulated genes involved in antioxidant defenses, inflammatory response and cell renewal. The collective antioxidant properties and transcriptional effect of this extract suggest potential antiaging properties which could be utilized in cosmetic and nutraceutical formulations.

Contribution of Chlorogenic Acids to the Inhibition of Human Hepatic Glucose-6-phosphatase Activity in Vitro by Svetol, a Standardized Decaffeinated Green Coffee Extract

Glucose-6-phosphatase (Glc-6-Pase) is a multicomponent system that exists primarily in the liver and catalyzes the terminal step in gluconeogenesis and glycogenolysis. Several studies have attempted to identify synthetic or natural compounds that inhibit this enzyme complex for therapeutic use in regulating blood glucose and type 2 diabetes. For this paper an in vitro structure-activity relationship study of several natural chlorogenic acids was conducted, and the active components of the natural decaffeinated green coffee extract Svetol were identified. Glucose-6-phosphate (Glc-6-P) hydrolysis was measured in the presence of Svetol or chlorogenic acids in intact human liver microsomes. Svetol significantly inhibited Glc-6-P hydrolysis in intact human liver microsomes in a competitive manner, and it was determined that chlorogenic acids (caffeoylquinic acids and dicaffeoylquinic acids) were the chief compounds mediating this activity. In addition, the structure-activity analysis showed that variation in the position of the caffeoyl residue is an important determinant of inhibition of Glc-6-P hydrolysis. This inhibition by Svetol contributes to its antidiabetic, glucose-lowering effects by reducing hepatic glucose production.

Carbon-14 biolabelling of wine polyphenols in Vitis vinifera cell suspension cultures

14C-L-phenylalanine is incorporated into a range of polyphenolic compounds when fed to grape cell cultures. Optimisation of several parameters such as the quantity of precursor applied and the duration of metabolism led to incorporation yields of 15% and to specific activities of 875 mu Ci g(-1) in stilbenes. Purification of the products by several chromatographic steps is reported. Both trans- and cis-resveratrols were easily obtained by enzymatic hydrolysis of their corresponding glucosides, with specific activity of 1200-1400 mu Ci g(-1). The specific radioactivity obtained for all the compounds is suitable for in vivo feeding trials to trace their metabolic fate when consumed by animals and for in vitro activity mechanism studies. Indeed, these polyphenols seem to be implicated in the health benefits associated with regular and moderate wine consumption but little is known about their pharmacokinetics and cellular uptake.

Obtaining Vitis Vinifera cell cultures producing higher amounts of malvidin-3-O-β-glucoside

To obtain a cell line of Vitis vinifera producing mainly malvidin-3-O-β-glucoside, three methods were studied: the initiation of new cell cultures from different varieties, the modification of phytohormones in the culture media of a high anthocyanin-producing cell line, and the selection by small-aggregate cloning of new cell lines from this high producing strain. The latter technique gave a cell line accumulating a malvidin-3-O-β-glucoside proportion of 63%.

DNA macroarray study of skin aging-related genes expression modulation by antioxidant plant extracts on a replicative senescence model of human dermal fibroblasts.

The formation of reactive oxygen species (ROS) is a widely accepted pivotal mechanism leading to skin aging. It increases with age, while the endogenous defense mechanisms that counter them decrease. This imbalance, called oxidative stress, leads to the progressive damage of cellular structures and results in accelerated aging. Antioxidant compounds can provide protection from endogenous and exogenous oxidative stress by scavenging free radicals. The main phenolic compounds of oak wood, mate leaf and benjoin resin antioxidant extracts were identified and the effects of these extracts on skin aging markers were evaluated using DNA macroarray technology. The transcriptional effect of the three antioxidant extracts was evaluated in vitro on a replicative senescence model of normal human dermal fibroblasts (NHDF), using a customized DNA macroarray specifically designed to investigate aging markers such as dermal structure, cell renewal, inflammatory response and oxidative stress mechanisms. Among the 149 genes detected, the three antioxidant extracts presented a significant regulation of five genes involved in inflammatory response, cell renewal and antioxidant defenses. The collective transcriptional effects of these extracts suggest interesting antiaging properties which could be utilized in nutraceutical antiaging formulations. Copyright