Myriam Richelle

The Biological Relevance of Direct Antioxidant Effects of Polyphenols for Cardiovascular Health in Humans Is Not Established

Human studies provide evidence for beneficial effects of polyphenol-rich foods on cardiovascular health. The antioxidant activity of polyphenols potentially explains these effects, but is the antioxidant activity a reliable predictor for these effects? An International Life Sciences Institute Europe working group addressed this question and explored the potential of antioxidant claims for polyphenols in relation to cardiovascular health by using the so-called Process for the Assessment of Scientific Support for Claims on Foods project criteria. In this process, analytical aspects of polyphenols, their occurrence in foods, dietary intake, and bioavailability were reviewed. Human studies on polyphenols and cardiovascular health were reviewed together with methods for biomarkers of oxidative damage and total antioxidant capacity (TAC). In retrospective studies, F2-isoprostanes and oxidized LDL, the most reliable biomarkers of lipid peroxidation, and measures for TAC showed the expected differences between cardiovascular disease patients and healthy controls, but prospective studies are lacking, and a causal relationship between these biomarkers and cardiovascular health could not be established. Therefore, the physiological relevance of a potential change in these biomarkers is unclear. We found limited evidence that some types of polyphenol-rich products modify these biomarkers in humans. A direct antioxidant effect of polyphenols in vivo is questionable, however, because concentrations in blood are low compared with other antioxidants and extensive metabolism following ingestion lowers their antioxidant activity. Therefore, the biological relevance of direct antioxidant effects of polyphenols for cardiovascular health could not be established. Overall, although some polyphenol-rich foods exert beneficial effects on some biomarkers of cardiovascular health, there is no evidence that this is caused by improvements in antioxidant function biomarkers (oxidative damage or antioxidant capacity).

Urinary isoprostane excretion is not confounded by the lipid content of the diet

This study aims to determine if isoprostanes accurately reflect in vivo lipid peroxidation or whether they are influenced by the lipid content of the diet. Isoprostanes were measured in urine of healthy subjects under different conditions of lipid intake and under conditions of oxidative stress (fasting). We found that isoprostanes were not influenced by the lipid content of the diet: the urinary level remained constant over 24 h as well as over 4 consecutive days when switching from high to low lipid intake. Urinary isoprostane excretion was increased by 40% following a 24 h fast. We concluded that urinary isoprostane excretion reflects endogenous lipid peroxidation in vivo.

Lycopene isomerisation takes place within enterocytes during absorption in human subjects.

Lycopene in fruits and vegetables occurs mostly (80-97 %) in the all-E configuration, whereas a considerable proportion of lycopene in the human body is present as Z-isomers. The Z-isomers offer potentially better health benefits and show improved antioxidant activity in vitro when compared with the all-E-isomer. The absorption of dietary lycopene is a complex process involving transfer of the carotenoid from the food matrix into micelles, uptake by enterocytes, packaging into chylomicrons and finally secretion into plasma. Isomerisation could take place at any of these individual steps. By exploiting in vitro and in vivo models, we traced lycopene isomerisation during absorption using various methods to mimic gastric and duodenal conditions, incorporation into mixed micelles, absorption and metabolism by various Caco-2 cell clones, and performed a postprandial study in human subjects to identify the profile of lycopene isomers in plasma chylomicrons. We demonstrate that all-E-lycopene remains unchanged during its passage in the gastrointestinal tract, including its incorporation into mixed micelles. The key site of lycopene isomerisation is inside the intestinal cells resulting in 29 % of lycopene as Z-isomers. Lycopene isomerisation in the various Caco-2 cell clones is consistent with that observed in human chylomicrons formed in a postprandial state. There is no selection in the release of lycopene isomers from enterocytes. Although there is a huge inter-individual variability of total lycopene absorption reported both in in vitro intestinal cell lines as well as in human chylomicrons, the lycopene isomer profile is quite similar.

The proportion of lycopene isomers in human plasma is modulated by lycopene isomer profile in the meal but not by lycopene preparation.

Dietary lycopene consists mostly of the (all-E) isomer. Upon absorption, (all-E) lycopene undergoes isomerisation into various (Z)-isomers. Because these isomers offer potentially better health benefits than the (all-E) isomer, the aim of the present study was to investigate if the profile of lycopene isomers in intestinal lipoproteins is affected by the profile of lycopene isomers in the meal and by the tomato preparation. Six postprandial, crossover tests were performed in healthy men. Three meals provided about 70 % of the lycopene as (Z)-isomers, either mainly as 5-(Z) or 13-(Z), or as a mixture of 9-(Z) and 13-(Z) lycopene, while three tomato preparations provided lycopene mainly as the (all-E) isomer. Consumption of the 5-(Z) lycopene-rich meal led to a high (60 %) proportion of this isomer in TAG-rich lipoproteins (TRL), indicating a good absorption and/or a low intestinal conversion of this isomer. By contrast, consumption of meals rich in 9-(Z) and 13-(Z) lycopene isomers resulted in a low level of these isomers but high amounts of the 5-(Z) and (all-E) isomers in TRL. This indicates that the 9-(Z) and 13-(Z) isomers were less absorbed or were converted into 5-(Z) and (all-E) isomers. Dietary (Z)-lycopene isomers were, therefore, differently isomerised and released in TRL during their intestinal absorption in men. Consuming the three meals rich in (all-E) lycopene resulted in similar proportions of lycopene isomers in TRL: 60 % (all-E), 20 % 5-(Z), 9 % 13-(Z), 2 % 9-(Z) and 9 % unidentified (Z)-isomers. These results show that the tomato preparation has no impact on the lycopene isomerisation occurring during absorption in humans.

β3-adrenoceptor agonist prevents alterations of muscle diacylglycerol and adipose tissue phospholipids induced by a cafeteria diet

BackgroundInsulin resistance induced by a high fat diet has been associated with alterations in lipid content and composition in skeletal muscle and adipose tissue. Administration of β3-adrenoceptor (β3-AR) agonists was recently reported to prevent insulin resistance induced by a high fat diet, such as the cafeteria diet. The objective of the present study was to determine whether a selective β3-AR agonist (ZD7114) could prevent alterations of the lipid profile of skeletal muscle and adipose tissue lipids induced by a cafeteria diet.MethodsMale Sprague-Dawley rats fed a cafeteria diet were treated orally with either the β3-AR agonist ZD7114 (1 mg/kg per day) or the vehicle for 60 days. Rats fed a chow diet were used as a reference group. In addition to the determination of body weight and insulin plasma level, lipid content and fatty acid composition in gastronemius and in epididymal adipose tissue were measured by gas-liquid chromatography, at the end of the study.ResultsIn addition to higher body weights and plasma insulin concentrations, rats fed a cafeteria diet had greater triacylglycerol (TAG) and diacylglycerol (DAG) accumulation in skeletal muscle, contrary to animals fed a chow diet. As expected, ZD7114 treatment prevented the excessive weight gain and hyperinsulinemia induced by the cafeteria diet. Furthermore, in ZD7114 treated rats, intramyocellular DAG levels were lower and the proportion of polyunsaturated fatty acids, particularly arachidonic acid, in adipose tissue phospholipids was higher than in animals fed a cafeteria diet.ConclusionsThese results show that activation of the β3-AR was able to prevent lipid alterations in muscle and adipose tissue associated with insulin resistance induced by the cafeteria diet. These changes in intramyocellular DAG levels and adipose tissue PL composition may contribute to the improved insulin sensitivity associated with β3-AR activation.

Gas chromatograhy-tandem mass spectrometry determination of 8-iso-PGF2α, a biomarker of in vivo lipid peroxidation, in human plasma and urine

The measurement of isoprostanes is a promising assay that is specific and sensitive enough to detect in vivo lipid peroxidation. We present here a gas chromatography-tandem mass spectrometry (GC/MS/MS)method that enables determination of 8-iso-prostaglandin F2α (8-iso-PGF2α)in human plasma and urine. After the addition of [2H4]-PGF2α as the internal standard to acidified plasma or urine, the samples are purified on C18 and silica cartridges, derivatised as pentafluorobenzyl esters, extracted with diethyl ether, purified on silica gel TLC plates and finally silylated. Then, 8-iso-PGF2α and its internal standard are measured by GC/MS/MS in selective-reaction monitoring mode using the transition [M −181]− to [M −181 – (3 × 90)]−. The detection limit of this method is 5 pg mL−1. Its application is presented in two situations of oxidative stress: in vitro low-density lipoprotein oxidation and in smokers. Measurement of urinary 8-iso-PGF2α levels provides a non-invasive in vivo index of free radical generation that appears not to be confounded by changes in diet.

Simultaneous determination of deuterated and non-deuterated α-tocopherol in human plasma by high-performance liquid chromatography

Labelled tocopherol is used to evaluate its absorption by biodiscriminating the dietary intake from the endogenous tocopherol pool of subject. A normal-phase high-performance liquid chromatographic method is described for the easy separation and quantification of deuterated (d(6)) and non-deuterated alpha-tocopherol. The alpha-tocopherol isotopomers were extracted from plasma triacylglycerol-rich lipoproteins in hexane, separated by two EC Nucleosil columns in series with a mobile phase of hexane-isopropanol (659.34:0.786, w/w) running isocratically. The detection of d(6)-alpha-tocopherol was performed by its UV absorbance at 297 nm with a limit of detection of 34 pmol/ml, a limit of quantification of 83 pmol/ml and a range of determination of 34-9905 pmol/ml. Between- and within-assay RSDs were 2.4% (n=10) and 2.7% (n=5), respectively.

Carotenoids and Skin

Dietary carotenoid bioavailability is rather low in comparison to other macronutrients. However, they are absorbed and distributed to hepatic and some extrahepatic tissues, including skin. Co-consumption of dietary fat is essential for carotenoid bioavailability. In contrast, the presence of some other dietary constituents, such as fiber, may reduce carotenoid bioavailability. Fruits and vegetables contain bioactive agents, including carotenoids, that protect not only plants but also humans against solar ultraviolet (UV) radiation damage. Only recently has the role of dietary bioactive agents in the photoprotection of skin started to be investigated and appreciated. Systemic photoprotection via a dietary supply of carotenoids may contribute significantly to skin health and complement the use of sunscreens in protecting the skin against the damaging effects of solar UV exposure. However, the knowledge acquired so far on the role of dietary carotenoids in photoprotection is still in its infancy and needs to be further investigated.

Bioavailability and Skin Bioefficacy of Vitamin C and E

Publisher Summary Skin is constantly exposed to pro-oxidant environmental stresses from an array of sources, such as air pollutants, solar UV light, chemical oxidants, micro-organisms, cigarette smoke, and ozone, which causes many changes in the skin. The most popular nutrients to alleviate skin changes such as aging, photosensitivity, and dryness are vitamins, carotenoids, polyphenols, and minerals. The blood continuously replenishes the skin with bioactives, which can then be distributed to all skin compartments, that is, epidermis, dermis, subcutaneous fat, and also sebum. This chapter presents the mechanisms involved in the bioavailability of nutrients such as vitamins C and E at the intestinal as well as at the skin level but also their bioefficacies in skin. Vitamin E has been shown to present an antioxidant property, and to play a role in photoprotection and in the prevention of aging, alone or in association with vitamin C. Literature on vitamin C for skin benefits is more abundant, showing evidence for the beneficial effect of this ingredient on dermal matrix formation, or epidermal differentiation, against UV-induced skin damage, and oxidative stress, indicating that vitamin C supplementation may be of interest to target skin aging, photoprotection, and skin xerosis. This chapter reviews current knowledge on the journey of dietary bioactives, that is, vitamin C and vitamin E from the mouth to skin, as well as their biological activities in the skin.