Transport et métabolisme du trans-resvératrol dans la lignée cellulaire d'hépatoblastome humain HepG2

Le trans-resvératrol, polyphénol naturel abondant dans le vin rouge pourrait contribuer à la lutte contre de nombreuses pathologies, dont le cancer. Pour une éventuelle utilisation thérapeutique, sa biodisponibilité au niveau hépatique a été étudiée sur le modèle HepG2, par microscopie de fluorescence, transport de resvératrol radiomarqué et analyse des milieux de culture par HPLC et spectrométrie de masse. Il a ainsi été montré que le resvératrol était rapidement capté, à la fois par diffusion passive et transport facilité, puis métabolisé sous forme de mono- et disulfates qui sont ensuite évacués par des transporteurs MRPs. Parallèlement, le transport et le pouvoir antiprolifératif de dérivés de resvératrol ont été étudiés et l'effet perturbateur endocrinien du resvératrol sur le transport de l'œstradiol a été comparé à celui d'autres œstrogénomimétiques (bisphénol A et génistéine).Trans-resveratrol, a natural polyphenol abundant in red wine could fight against numerous diseases such as cancer. To consider a therapeutic use of resveratrol, its bioavailability at hepatic level was studied with HepG2 cell line by using fluorescence microscopy, radiolabelled resveratrol transport determination and analysis of cell culture media by HPLC and mass spectrometry. It was shown that resveratrol was rapidly taken up by cells following both an active transport process and a passive transport before its modification in mono- and disulfated metabolites that were finally excreted by MRPs transporters. At the same time on this cell line, transport and antiproliferative properties of resveratrol derivatives were compared. Finally, the endocrine disrupting effect of resveratrol and other estrogenomimetics (genistein and bisphenol A) was studied at the estradiol transport level.DIJON-BU Sciences Economie (212312102) / SudocSudocFranceF

Resveratrol: preventing properties against vascular alterations and ageing.

International audienceCardiovascular diseases are the leading cause of death in developed countries where the common pathological substrate underlying this process is atherosclerosis. Several new concepts have emerged in relation to mechanisms that contribute to the regulation of the vascular diseases and associated inflammatory effects. Recently, potential antioxidants (vitamin E, polyphenols) have received much attention as potential anti-atherosclerotic agents. Among the polyphenols with health benefic properties, resveratrol, a phytoalexin of grape, seem to be a good candidate protecting the vascular walls from oxidation, inflammation, platelet aggregation, and thrombus formation. In this review, we focus on the mechanism of resveratrol cardiovascular benefic effects. We analyze, in relation with the different steps of atherosclerotic process, the resveratrol properties at multiple levels, such as cellular signaling, enzymatic pathways, apoptosis, and gene expression. We show and discuss the relationship with reactive oxygen species, regulation of pro-inflammatory genes including cycloxygenases and cytokines in molecular inflammatory and aging processes, and how the regulation of these activites by resveratrol can lead to a prevention of vascular diseases

Resveratrol as a chemopreventive agent: a promising molecule for fighting cancer.

International audienceResveratrol (3,4',5 tri-hydroxystilbene) is a phytoalexin produced in hudge amount in grapevine skin in response to infection by Bothrytis cinerea. This production of resveratrol blocks the proliferation of the pathogen, thereby acting as a natural antibiotic. Numerous studies have reported interesting properties of trans-resveratrol as a preventive agent against important pathologies i.e. vascular diseases, cancers, viral infection or neurodegenerative processes. Moreover, several epidemiological studies have revealed that resveratrol is probably one of the main microcomponents of wine responsible for its health benefits such as prevention of vaso-coronary diseases and cancer. Resveratrol acts on the process of carcinogenesis by affecting the three phases: tumor initiation, promotion and progression phases and suppresses the final steps of carcinogenesis, i.e. angiogenesis and metastasis. It is also able to activate apoptosis, to arrest the cell cycle or to inhibit kinase pathways. Interestingly, resveratrol does not present any cytotoxicity in animal models. Moreover, concentrations of resveratrol in blood seem to be sufficient for anti-invasive activity. The enterohepatic recirculation may contribute to a delayed elimination of the drug from the body and bring about a prolonged effect. By its binding to plasmatic proteins, resveratrol also exhibits a prolonged effect. Interestingly, low doses of resveratrol can sensitize to low doses of cytotoxic drugs and so provide an innovative strategy to enhance the efficacy of anticancer therapy in various human cancers. By these properties, resveratrol appears to be a good candidate in chemopreventive or chemotherapeutic strategies and is believed to be a novel weapon for new therapeutic strategies

Nutritional prevention by dietary resveratrol as chemopreventive agent: application to colorectal cancers

Resveratrol (3,4',5 tri-hydroxystilbene) is a plant phytoalexin produced in massive amount in grapevine skin in response to stress such as UV, phytosanitary treatment, and mostly following infection by Bothrytis cinerea. In this later case, the production of resveratrol inhibits the proliferation of the pathogen, thereby acting as a natural antifungal. Many experimental studies have reported interesting properties of trans-resveratrol as a preventive agent against important pathologies i.e. vascular diseases, cancers, viral infection or neurodegenerative processes. In addition, several epidemiological studies indicated that resveratrol would be the main microcomponent of wine leading health benefits such as prevention of vaso-coronary diseases and cancer (so called the "French paradox"). Resveratrol prevents (or delays) carcinogenesis by inhibiting the three phases of cancer process: initiation, promotion, progression and invasion phases. It also exhibits pro-apoptic properties especially towards colorectal cancers. Importantly, resveratrol is not toxic in animal models even at high dosage. Moreover, plasmatic concentrations of resveratrol would to be sufficient for anti-invasive activity. The enterohepatic blood recirculation contributes to a delayed elimination of the molecule from the body which can also show a prolonged effect enhanced by its binding to plasmatic proteins. Interestingly resveratrol can sensitize to low doses of cytotoxic drugs and so provide new approaches to enhance the efficacy of anticancer therapy in human cancers

Endocrine disruptors found in food modulate estradiol cellular metabolism

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Antiproliferative activities of resveratrol and related compounds in human hepatocyte derived HepG2 cells are associated with biochemical cell disturbance revealed by fluorescence analyses.

International audienceResveratrol is a well known polyphenol largely produced in grapevine. It is a strong antioxidant and a free radical scavenger. It exhibits several beneficial effects for health including cancer. Resveratrol antioxidant activity is essential in the prevention of chemical-induced cancer by inhibiting initiation step of carcinogenesis process but it is also considered to inhibit cancer promotion and progression steps. While the effects of resveratrol on cancer cells are widely described, the data available on the antiproliferative potential of resveratrol derivatives remain weak. Nevertheless, resveratrol analogs could exhibit stronger potentials than the parent molecule. So, we compared the cellular effects of trans-resveratrol, trans-epsilon-viniferin and their respective acetate derivatives, as well as a polyphenol mixture extracted from grapevine shoots, called vineatrol. We studied their abilities to interfere with cell proliferation, their uptake and their effects on parameters of cellular state in human hepatoma cells (HepG2). Cell growth experiments show that resveratrol triacetate presents a slightly better antiproliferative potential than resveratrol. The dimer epsilon-viniferin,as well as its pentaacetate analog, is less powerful than resveratrol, although a similar uptake kinetics in cells. Interestingly, among the tested polyphenols, vineatrol is the most potent solution, indicating a possible synergistic effect of both resveratrol and epsilon-viniferin. We took advantage of the fluorescence properties of these compounds to evidence cellular uptake by using flow cytometry. In addition, by competition assay, we demonstrate that resveratrol triacetate enters in hepatic HepG2 cells by the same way as resveratrol. By autofluorescence in situ measurement we observed that resveratrol and related compounds induce deep changes in cells activity. These changes occur mainly by increasing NADPH cell content and the number of green fluorescent cytoplasmic granular structures which may be related to an induction of detoxifying enzyme mechanisms

Major sulfation pathway for resveratrol metabolism in human hepatic derived cells and resveratrol –dependent inducibility of conjugating enzymes. Perspective in nutritional prevention

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Resveratrol in human hepatoma HepG2 cells: metabolism and inducibility of detoxifying enzymes.

trans-Resveratrol is a polyphenol present in several plant species. Its chemopreventive properties against several diseases have been largely documented. To validate a model for the study of the factors influencing its biological fate at the hepatic level, the metabolism and the efflux of resveratrol were studied in the human hepatoblastoma cell line, HepG2. Comparative high-performance liquid chromatography analysis of cell culture media before and after deconjugation showed that resveratrol was rapidly conjugated; at the concentration of 10 microM, it was entirely metabolized at 8 h of incubation. Two main resveratrol metabolites, monosulfate and disulfate, were identified by atmospheric pressure chemical ionization-mass spectrometry, thanks to their quasi-molecular ion and their characteristic fragmentation. To correlate with the auto-induction of resveratrol metabolism evidenced in HepG2 cells after a pretreatment for 48 h with 10 microM resveratrol, the inducibility of phase II enzymes by resveratrol was studied by real-time quantitative reverse transcriptase-polymerase chain reaction and flow cytometry. Observed, in particular, were an increase in mRNA expression levels of three metabolizing enzymes, two isoforms of UDP-glucuronosyltransferases, UGT1A1 and UGT2B7 (5-fold increased), and a sulfotransferase, ST1E1, in cells pretreated for 24 h with 10 microM resveratrol. These results were correlated with an increase in protein expression, especially after 48 h of treatment. On the other hand, the intracellular resveratrol retention in cells treated with MK571 (3-[[3-[2-(7-chloroquinolin-2-yl)vinyl]phenyl]-(2-dimethylcarbamoylethylsulfanyl)methylsulfanyl] propionic acid), a multidrug resistance-associated protein inhibitor, strongly suggests the involvement of this ABC transporter family in the efflux of resveratrol conjugates from human liver

Major sulfation pathway for resveratrol metabolism in human hepatic derived cells and resveratrol –dependent inducibility of conjugating enzymes. Perspective in nutritional prevention

International audienc

Endocrine disruptors found in food modulate estradiol cellular metabolism

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