Biochemical and Biophysical Research Communications
Human hepatic cell uptake of resveratrol: involvement of both passive diffusion and carrier-mediated process
Section snippets
Materials and methods
Chemicals. Trans-resveratrol was purchased from Sigma (St. Louis, MO, USA). [3H]-trans-resveratrol (specific activity: 74 GBq/mmol) labelled in ortho and para of benzenic rings was prepared for us by Amersham. Stock solutions of trans-resveratrol were prepared in absolute ethanol or in dimethyl sulfoxide (DMSO), wrapped in aluminium foil for protection against light, and stored at 4 °C when not in use. Resveratrol 3,5,4′-triacetate and ε-viniferin were a gift from Actichem (Montauban, Fr). Bovine
Fluorescence measurement of resveratrol uptake
Slides were prepared from HepG2 cells preincubated with resveratrol during various times and then fixed. They were observed by fluorescence microscopy (Fig. 1). Resveratrol intrinsic fluorescence allowed a direct visualization of its intracellular uptake. A green fluorescence was observed in treated cells, markedly higher than basal fluorescence of untreated cells. Green fluorescence was obvious after incubation times less than 2 min. Fluorescence appeared to be distributed among the whole cell.
Discussion
For the first time, we report that the fluorescent properties of resveratrol allowed an easy method to follow the transport of this polyphenol by fluorescence microscopy. Such a method is also convenient for the study of the transport of resveratrol derivatives such as resveratrol triacetate and ε-viniferin (data not shown).
The observation of fluorescent slides showed that resveratrol is present essentially in cytoplasm. The nucleolar localization of resveratrol remains to be explained. A
Acknowledgements
This study was supported by the “Conseil Régional de Bourgogne,” BIVB, IREB, ONIVINS, and the “Ligue Bourguignonne contre le Cancer.” Thanks to Mr. O. Bocrie for preliminary experiments.
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Cited by (0)
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The first two authors have equally contributed to this work.
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Present address: Pharmakokinetic and Metabolism, Sanofi Synthélabo Recherche, 91380 Chilly-Marzarin, France.