Properties of a fluorescent bezafibrate derivative (DNS-X). A new tool to study peroxisome proliferation and fatty acid beta-oxidation.
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 beta-oxidation, lipoprotein structure [apolipoprotein (apo) A2, apo CIII], 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-dimethylaminonaphthalene-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 PPAR alpha activity as well as other peroxisome proliferators do.
Berlot
J P
JP
Université de Bourgogne, Laboratorie de Biologie Moléculaire et Cellulaire, Faculté des Sciences Gabriel, Dijon, France.
Lutz
T
T
Cherkaoui Malki
M
M
Nicolas-Frances
V
V
Jannin
B
B
Latruffe
N
N
D000818:Animals; D001629:Bezafibrate / Q000031:analogs & derivatives / Q000737:chemistry* / Q000494:pharmacology; D002455:Cell Division; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003600:Cytosol / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006529:Hepatomegaly; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D008956:Models, Chemical; D010088:Oxidoreductases / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D020025:Peroxisome Proliferators / Q000378:metabolism; D020675:Peroxisomes / Q000201:enzymology; D010957:Plasmids; D051381:Rats; D017208:Rats, Wistar; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D013997:Time Factors; D017186:Titrimetry; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D005227:Fatty Acids; D005456:Fluorescent Dyes; D020025:Peroxisome Proliferators; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors; C426804:fluorescent bezafibrate; D010088:Oxidoreductases; C028427:palmitoyl CoA oxidase; D010100:Oxygen; D001629:Bezafibrate