The effect of dexamethasone, insulin and triiodothyronine on microsomal NADPH-cytochrome-c (P-450) reductase in primary cultures of isolated hepatocytes.
NADPH-cytochrome-c (P-450) reductase, a flavoprotein, is a constituent of the hepatic microsomal polysubstrate monooxygenase and catalyzes the transfer of electrons from NADPH to cytochrome P-450. The hormonal regulation of NADPH-cytochrome-c reductase activity and protein has been examined in insolated hepatocytes cultured as monolayers for 48 h in Waymouth's MB752/1 medium fortified with insulin, dexamethasone and triiodothyronine. No similarity between the response of NADPH-cytochrome-c reductase and of tyrosine aminotransferase and malate dehydrogenase activity to dexamethasone and triiodothyronine treatment could be detected. In the absence of hormones about 65% of the original NADPH-cytochrome-c reductase activity and protein estimated by the immunochemical staining technique was retained. Culture of hepatocytes in insulin (10.0 mU/ml) or dexamethasone (100 nM) alone but not triiodothyronine improved the retention of reductase activity and protein. Only when hepatocytes were cultured in insulin, triiodothyronine and dexamethasone could NADPH-cytochrome-c reductase activity and protein be maintained at the original level. Dexamethasone alone was found to enhance consistently retention of reductase protein, but not reductase activity, to approximately the same level as in freshly isolated hepatocytes. The results suggest that microsomal NADPH-cytochrome-c reductase activity and protein can be maintained in isolated hepatocytes at the original level by culturing the cells in dexamethasone, insulin and triiodothyronine.
D000818:Animals; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D007118:Immunoassay; D005779:Immunodiffusion; D007328:Insulin / Q000494:pharmacology*; D008291:Malate Dehydrogenase / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D014284:Triiodothyronine / Q000494:pharmacology*; D014444:Tyrosine Transaminase / Q000378:metabolism
D007328:Insulin; D014284:Triiodothyronine; D003907:Dexamethasone; D008291:Malate Dehydrogenase; D009251:NADPH-Ferrihemoprotein Reductase; D014444:Tyrosine Transaminase