Lipid metabolic changes caused by short-chain ceramides and the connection with apoptosis.
The effects of the short-chain ceramides D-erythro-N-acetylsphingosine (C(2)-ceramide), 6-[N-(7-nitrobenz-2-oxa-1, 3-diazole-4-yl)amino]hexanoyl-D-erythro-sphingosine(NBD-ceramide) and N-[4,4-difluoro-5,7-dimethyl-4-bora-3a, 4a-diaza-s-indacene-3-pentanoyl]-D-erythro-sphingosine (DMB-ceramide) on the incorporation of [(14)C]acetate into baby-hamster kidney (BHK) fibroblasts have been examined. C(2)-ceramide at concentrations up to 20 microM caused an inhibition of synthesis of phosphatidylcholine (PtdCho), sphingolipids and cholesterol within 2 h. Similar effects in BHK cells were seen using other radioactive tracers ([(3)H]water, [(3)H]palmitate and [(3)H]choline) and using HL60 cells labelled with [(14)C]acetate. The inhibition of PtdCho synthesis corresponded to an accumulation of label in diacylglycerol and triacylglycerol, probably as a consequence of cytidylyltransferase blockade. With [(3)H]choline label, the decrease in sphingomyelin synthesis could be partly accounted for by accumulation of a slow-moving lipid, likely to be C(2)-sphingomyelin. NBD-ceramide also reduced sphingomyelin and cholesterol biosynthesis, but had much less effect on PtdCho and acylglycerols. In contrast, the only apparent effect of DMB-ceramide was to inhibit synthesis of sphingomyelin, with a reciprocal increase in DMB-sphingomyelin synthesis. However, all of these short-chain ceramides caused massive apoptosis after 18 h, whereas addition of N-acetyldihydrosphingosine or elevation of natural ceramide by treatment of cells with sphingomyelinase had little effect on lipid synthesis or apoptosis. The present findings suggest that the apoptotic effect of short-chain ceramides is sometimes associated with inhibition of cytidylyltransferase, but is more closely correlated with a competitive inhibition of normal sphingomyelin biosynthesis.
Allan
D
D
Department of Physiology, University College London, University Street, London WC1E 6JJ, U.K. d.allan@ucl.ac.uk
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001896:Boron Compounds / Q000378:metabolism / Q000494:pharmacology; D002460:Cell Line; D002518:Ceramides / Q000378:metabolism / Q000494:pharmacology*; D002784:Cholesterol / Q000096:biosynthesis; D006224:Cricetinae; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D050356:Lipid Metabolism; D010713:Phosphatidylcholines / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism; D013107:Sphingolipids / Q000378:metabolism; D013109:Sphingomyelins / Q000096:biosynthesis; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology
D001896:Boron Compounds; D002518:Ceramides; C405489:N-(4,4-difluoro-5,7-dimethyl-4--bora-3a,4a-diaza-s-indacene-3-pentanoyl)sphingosine; C064769:N-acetylsphingosine; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D013107:Sphingolipids; D013109:Sphingomyelins; C037974:N-(7-(4-nitrobenzo-2-oxa-1,3-diazole))-6-aminocaproyl sphingosine; D002784:Cholesterol; D009327:4-Chloro-7-nitrobenzofurazan; D013110:Sphingosine