Docosahexaenoic acid down-regulates phenobarbital-induced cytochrome P450 2B1 gene expression in rat primary hepatocytes via the sphingomyelinase/ceramide pathway.
Docosahexaenoic acid (DHA) regulates the expression of cytochrome P450 2B1 (CYP 2B1) in rat primary hepatocytes in response to xenobiotics. Ceramide, a lipid signaling molecule, is involved in various physiological processes and can be generated by the hydrolysis of sphingomyelin via sphingomyelinase (SMase). DHA activates SMase and increases ceramide formation in vitro. Ceramides differentially enhance adenylyl cyclase activity in vitro depending on the chain length of their fatty acids. In addition, the cAMP-dependent PKA pathway down-regulates CYP 2B1 expression induced by phenobarbital (PB). In the present study, we determined the effect of DHA on SMase transactivation and the downstream pathway in CYP 2B1 expression induced by PB. SMase was activated by DHA 2 h after treatment, and D609 (an SMase inhibitor) attenuated the inhibition of PB-induced CYP 2B1 expression by DHA. Ceramide formation reached a maximum 3 h after DHA administration. C2-ceramide dose-dependently inhibited PB-induced CYP 2B1 expression and increased intracellular cAMP concentrations. SQ22536 (an adenylyl cyclase inhibitor) and H89 (a PKA-specific inhibitor) partially reversed the inhibition of PB-induced CYP 2B1 expression by C2-ceramide. These results suggest that stimulation of SMase, generation of ceramide and activation of the cAMP-dependent PKA pathway are involved in the inhibition exerted by DHA.
Lu
Chia-Yang
CY
Department of Nutrition, Chung Shan Medical University, Taichung 40201, Taiwan.
Li
Chien-Chun
CC
Liu
Kai-Li
KL
Tsai
Chia-Wen
CW
Lii
Chong-Kuei
CK
Chen
Haw-Wen
HW
D000067956:Adenylyl Cyclase Inhibitors; D000818:Animals; D002478:Cells, Cultured; D002518:Ceramides / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D019362:Cytochrome P-450 CYP2B1 / Q000235:genetics / Q000378:metabolism*; D004281:Docosahexaenoic Acids / Q000008:administration & dosage*; D015536:Down-Regulation; D022781:Hepatocytes / Q000378:metabolism; D008297:Male; D010634:Phenobarbital / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D013108:Sphingomyelin Phosphodiesterase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism; D013997:Time Factors; D015533:Transcriptional Activation
D000067956:Adenylyl Cyclase Inhibitors; D002518:Ceramides; C064769:N-acetylsphingosine; D004281:Docosahexaenoic Acids; D000242:Cyclic AMP; D019362:Cytochrome P-450 CYP2B1; D017868:Cyclic AMP-Dependent Protein Kinases; D013108:Sphingomyelin Phosphodiesterase; D013110:Sphingosine; D010634:Phenobarbital