Ascorbic acid deficiency and cytochrome P-450 in adult rat hepatocytes in primary monolayer culture. |
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Authors: | D M Bissell P S Guzelian |
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Affiliation: | 1. Department of Medicine and the Liver Center, University of California, San Francisco, California, U.S.A.;2. The Department of Medicine, Medical College of Virginia, Richmond, Virginia U.S.A. |
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Abstract: | Adult rat hepatocytes in primary monolayer culture exhibit selective alteration of microsomal constituents and functions during the first hours of incubation ex vivo, including a striking decrease in the concentration of cytochrome P-450. The present studies document that these alterations are due in part to deficiency of l-ascorbate in the cultured cells. The deficiency appears to develop both by loss of the vitamin from the cells during their preparation and by a diminished synthetic capacity for ascorbate. Supplementation of the culture medium with l-ascorbate, at a concentration sufficient to restore intracellular levels of vitamin C to normal, results in maintenance of significantly increased concentrations of cytochromes P-450 and b5. The activity of NADPH cytochrome c reductase similarly is ascorbate-dependent, suggesting that the vitamin plays a role in the formation and/or stabilization of membrane protein or lipid. Microsomal heme metabolism appeared to be unaffected by the presence or absence of ascorbate. |
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Keywords: | Recipient of Research Career Development Award K04-GM-00149 from the National Institutes of Health to whom correspondence should be addressed at Room 5H7, Department of Medicine, San Francisco General Hospital, San Francisco, California 94110. |
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