The role of phosphatidylinositol 3-kinase, Src kinase, and protein kinase A signaling pathways in insulin and glucagon regulation of CYP2E1 expression.
The signaling pathways involved in insulin and glucagon regulation of CYP2E1 expression were examined in primary cultured rat hepatocytes. Insulin addition to primary cultured rat hepatocytes for 24 h resulted in an approximately 80% and >90% decrease in CYP2E1 mRNA levels at 1 and 10 nM insulin, respectively, relative to untreated cells. Addition of the phosphatidylinositol 3-kinase inhibitor wortmannin, or the Src kinase inhibitor geldanamycin, prior to insulin addition, inhibited the insulin-mediated decline in CYP2E1 mRNA. In contrast, treatment of cells with glucagon (100 nM), or the cAMP analogue dibutyryl-cAMP (50 microM), for 24 h increased CYP2E1 mRNA levels by approximately 7-fold. Addition of the protein kinase A inhibitor H89 prior to glucagon treatment attenuated the glucagon-mediated increase in CYP2E1 mRNA by approximately 70%. Glucagon (100 nM) opposed the effects of insulin (1 nM) on CYP2E1 mRNA expression and conversely, insulin blocked the effects of glucagon. These data provide compelling evidence for the regulation of CYP2E1 expression via mutually antagonistic signaling pathways involving insulin and glucagon.
Woodcroft
K J
KJ
Institute of Chemical Toxicology, Wayne State University, Detroit, Michigan 48201, USA.
Novak
R F
RF
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D016227:Benzoquinones; D003994:Bucladesine / Q000494:pharmacology; D002478:Cells, Cultured; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D019392:Cytochrome P-450 CYP2E1 / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005934:Glucagon / Q000494:pharmacology*; D007328:Insulin / Q000494:pharmacology*; D047029:Lactams, Macrocyclic; D008099:Liver / Q000201:enzymology; D008297:Male; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D011809:Quinones / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D000077191:Wortmannin; D019061:src-Family Kinases / Q000378:metabolism*
D000730:Androstadienes; D016227:Benzoquinones; D004791:Enzyme Inhibitors; D005419:Flavonoids; D007328:Insulin; D047029:Lactams, Macrocyclic; D011809:Quinones; D012333:RNA, Messenger; D003994:Bucladesine; D005934:Glucagon; D019392:Cytochrome P-450 CYP2E1; D019869:Phosphatidylinositol 3-Kinases; D019061:src-Family Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D000077191:Wortmannin; C001277:geldanamycin