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Differential alterations of ethanolamine and choline phosphoglyceride metabolism by clofibrate and retinoic acid in human fibroblasts are not mediated by phorbol ester-sensitive protein kinase C

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Lipids

Abstract

Peroxisomal proliferators and retinoids have been reported to interact to regulate lipid metabolism, particularly β-oxidation of fatty acids. Based on postulated interactions of these agents at the levels of receptors and response elements, we examined whether interactions exist between the peroxisomal proliferator, clofibrate (CLF), and retinoic acid (RA) in modulation of phospholipid turnover in cultured human skin fibroblasts. Treatment of cultured cells with either 25 μM CLF or 1 μM RA alone decreased [14C]ethanolamine incorporation into ethanolamine phosphoglycerides (EPG) by 20–30%, and simultaneous exposure to both agents resulted in additive inhibition. By contrast, [3H]choline incorporation into phospholipid was stimulated 5–30% by incubation with either agent; when CLF and RA were administered together, the stimulatory effects were additive. Different types of pulse-chase studies examining effects on uptake, biosynthesis, and degradation of labelled phospholipids indicated stimulation of EPG degradation and inhibition of phosphatidylcholine degradation by CLF; no effect on catabolism of either phospholipid was observed with RA. Combinations of modifiers of protein kinase activity [4β-12-O-tetradecanoylphorbol-13-acetate (β-TPA), 1-(5-isoquino-linesulfonyl)-2-methylpiperazine dihydrochloride,N-(2′-guanidinoethyl)-5-isoquinolinesulfonamide hydrochloride,bis-indolylmaleimide, staurosporine] indicated that β-TPA-responsive protein kinases were not involved. Accordingly, CLF and RA regulate biosynthesis and degradation of ethanolamine and choline phosphoglycerides in cultured skin fibroblasts by different mechanisms that do not involve classical protein kinase C (PKC) isoforms, even though turnover of phospholipids generating lipid activators of PKC occurs.

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Abbreviations

CLF:

clofibrate

DMSO:

dimethylsulfoxide

EPG:

ethanolamine phosphoglycerides

HA-1004:

N-(2′-guanidinoethyl)-5-isoquinolinesulfonamide hydrochloride

H-7:

1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride

MEM:

Eagle's minimal essential medium

PKA:

protein kinase A

PKC:

protein kinase C

PLC:

phospholipase C

PLD:

phospholipase D

PMSF:

phenylmethylsulfonylfluoride

PPAR:

peroxisomal proliferator-activated receptor(s)

PtdCho:

phosphatidylcholine

RA:

retinoic acid

β-TPA:

β-12-O-tetradecanoylphorbol-13-acetate

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  1. Atlantic Research Centre, Dalhousie University, 5849 University Ave., B3H 4H7, Halifax, Nova Scotia, Canada

    Suzan G. Mandla, David M. Byers, Neale D. Ridgway & Harold W. Cook

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  1. Suzan G. Mandla
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Mandla, S.G., Byers, D.M., Ridgway, N.D. et al. Differential alterations of ethanolamine and choline phosphoglyceride metabolism by clofibrate and retinoic acid in human fibroblasts are not mediated by phorbol ester-sensitive protein kinase C. Lipids 31, 747–755 (1996). https://doi.org/10.1007/BF02522891

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