Studies on the maintenance of cytochromes P‐450 and b 5, monooxygenases and cytochrome reductases in primary cultures of rat hepatocytes

FEBS Letters

Melvin, W.T.²  Burke, M.D.¹  Grant, M.H.³  Melvin, M.A.L.²  Shaw, P.¹ 

[1] Department of Pharmacology, University of Aberdeen, Aberdeen AB9 2ZD, Scotland;Departmentof Biochemistry, University of Aberdeen, Aberdeen AB9 2ZD, Scotland;Department of Therapeutics and Clinical Pharmacology, Aberdeen AB9 2ZD, Scotland

关键词: Cultured hepatocyte;    Cytochrome P-450;    Mixed-function oxidase;    NADPH-cytochrome c reductase;    NADH-cytochrome b5 reductase;    EROD;    O-deethylation of ethoxyresorufin;    PPOD;    O-depentylation of pentoxyphenoxazone;   

DOI  :  10.1016/0014-5793(85)80436-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The cytochrome P-450 content of rat hepatocytes declined rapidly over 72 h in culture, due primarily to denaturation to cytochrome P-420. Six different media were investigated for their ability to conserve cytochrome P-450 during culture, and the most successful was a modified Earle's medium. After 72 h culture in this medium, cytochromes P-450 and b ₅, NADH-cytochrome b ₅- and NADPH-cytochrome c-reductases were maintained at 40, 100, 35 and 52% of fresh cell values, respectively. Cytochrome P-450 showed differential functional stability during culture with ethoxyresorufin O-deethylation being more stable than either pentoxyphenoxazone O-depentylation or biphenyl 4-hydroxylation. Monooxygenase activities declined faster than did cytochrome P-450 content. This discrepancy was not explained by loss of the flavin nucleotides, FMN or FAD.

【 授权许可】

Unknown