Changes in the concentration of seven forms of cytochrome P-450 in primary cultures of adult rat hepatocytes.

摘要: We prepared primary monolayer cultures of adult rat hepatocytes and measured the losses cytochromes P-450 with use specific antibodies directed against purified forms hepatic cytochrome which predominate in untreated rats (P-450UT-A, P-450UT-F) or treated phenobarbital (P-450PB-B/D, P-450PB-C, P-450PB/PCN-E) 3-methylcholanthrene (P-450 beta NF-B, NF/ISF-G). In from an incubated control medium, total P-450, spectrally as CO-binding hemoprotein, declined 68% during first 72 hr culture. However, sum immunoreactive only 24%, indicating that loss heme rather than protein accounts for much well-known hepatocyte cultures. 3-methylcholanthrene, individual at markedly differing rates. Incubation three different media previously reported to maintain levels failed prevent decline 24 medium containing metyrapone, level holocytochrome was maintained initial value hr, apparently by preventing net increasing concentrations P-450PB/PCN-E NF-B. Medium nicotinamide increased proportion NF-B relative other whereas cysteine-free P-450UT-F. conclude cultured involves its moiety coupled changes forms. Recognition these influenced components culture is important when using study xenobiotic metabolism toxicity liver.