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Properties of a fluorescent bezafibrate derivative (DNS-X). A new tool to study peroxisome proliferation and fatty acid β-oxidation

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Lipids

Abstract

The first peroxisome proliferator-activated receptor (PPAR) was cloned in 1990 by Issemann and Green. Many studies have reported the importance of this receptor in the control of gene expression of enzymes involved in lipid metabolic pathways including mitochondrial and peroxisomal fatty acid β-oxidation, lipoprotein structure [apolipoprotein (apo) A2, apo Clll], and fatty acid synthase. By using radiolabeled molecules, it was shown that peroxisome proliferators bind and activate PPAR. As an alternative method, we developed a fluorescent dansyl (1-dimethyl-aminonaphthalene-5-sulfonyl) derivative peroxisome proliferator from bezafibrate (DNS-X), a hypolipidemic agent that exhibits an in vitro peroxisome proliferative activity on rat Fao-hepatic derived cultured cells. However, until now, the effect of this new compound on the liver of animals and subcellular localization was unknown. In addition to in vivo rat studies, we present a more efficient large-scale technique of DNS-X purification. Treating rats (DNS-X in the diet at 0.3% w/w) for 6 d leads to a hepatomegaly and a marked increase in liver peroxisomal palmitoyl-CoA oxidase activity. We also developed a method to localize and quantify DNS-X in tissues or cell compartment organelles. The primarily cytosolic distribution of DNS-X was confirmed by direct visualization using fluorescence microscopy of cultured Fao cells. Finally, transfection assay demonstrated that DNS-X enhanced the PPARα activity as well as other peroxisome proliferators do.

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Abbreviations

Bezafibrate:

2-{4-[2-(4-chlorobenzamido)ethyl]phenoxy}-2-methylpropanoic acid

DNS-Cl:

5-(dimethylamino)naphthalene-1-sulfonylchloride, or dansyl chloride

DNS-OH:

5-(dimethylamino)naphthalene-1-sulfonic acid

DNS-Phe:

Dansyl phenylalanine

DNS-X:

2-[4-(N-dansyl-2-aminoethyl)phenoxy]-2-methylpropanoic acid

Fao cells:

a rat hepatoma-derived cell line

HPLC:

High-performance liquid chromatography

PBS:

phosphate-buffered saline

PP:

peroxisome proliferator

PPARα:

peroxisome proliferator-activated receptor α

PPRE:

peroxisome proliferator response element

RP-HPLC:

reversed-phase HPLC

TLC:

thin-layer chromatography

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Authors and Affiliations

  1. Faculté des Sciences Gabriel, Université de Bourgogne, LBMC, 6, Bd Grabriel, 21000, Dijon, France

    Jean-Pierre Berlot, Thomas Lutz, Mutaspha Cherkaoui Malki, Val rie Nicolas-Frances, Brigitte Jannin & Norbert Latruffe

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  1. Jean-Pierre Berlot
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  2. Thomas Lutz
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  3. Mutaspha Cherkaoui Malki
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  4. Val rie Nicolas-Frances
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  5. Brigitte Jannin
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  6. Norbert Latruffe
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Correspondence to Norbert Latruffe.

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Berlot, JP., Lutz, T., Malki, M.C. et al. Properties of a fluorescent bezafibrate derivative (DNS-X). A new tool to study peroxisome proliferation and fatty acid β-oxidation. Lipids 35, 1397–1404 (2000). https://doi.org/10.1007/s11745-000-0657-0

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  • DOI: https://doi.org/10.1007/s11745-000-0657-0

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