title: Studies on the metabolism of metallothionein and alkaline phosphatase of adult rat primary hepatocyte cultures: role of fetal calf serum and agonists of the phosphoinositide cascade.
abstract: Adult rat primary hepatocytes maintained in DMEM/F12 (Ham) media were used as a model system for studying the role of fetal calf serum (FCS) and agonists of the phosphoinositide cascade in the metabolism of metallothionein (MT) and alkaline phosphatase (ALP). Experiments were performed both after a 24 h preincubation with FCS and with bovine serum albumin (BSA). Hepatocytes were treated with dexamethasone (DEX), zinc (Zn) and with the agonists of the phosphoinositide cascade A23187, 1,2-dioctanoyl-sn-glycerol (DiC8), 12-O-tetradecanoylphorbol-13-acetate (TPA), angiotensin II (AT), platelet activating factor (PAF), Arg8-vasopressin (VP) and were analyzed for MT and ALP activity in cell homogenates. Cell viability was evaluated by lactate dehydrogenase (LDH) liberation into culture medium, induction of tyrosine aminotransferase (TAT) through DEX and by trypan blue exclusion. Overall, cell viability was improved by the FCS pretreatment and by DEX. Exposure of hepatocytes to the established direct inducers Zn and DEX of MT resulted in a manifold increase in MT, independent of whether the cultures were FCS pretreated or not. The FCS preincubation produced a moderate elevation of ALP activity by stimulating cell viability. However, ALP was unaltered in response to Zn and DEX. None of the experiments conducted with agonists of the phosphoinositide cascade led to an elevation of MT and ALP. Only the incubation of hepatocytes with A23187 resulted in a concentration dependent significant decrease of MT and ALP. This observation was due to a cytotoxic effect of A 23187, displayed by LDH leakage and an increase in the number of cells stained with trypan blue. In conclusion, in primary hepatocyte cultures agonists of the phosphoinositide did not have an effect on the metabolism of MT and ALP. Previous in vivo results indicating alterations of Zn metabolism in liver, therefore seem to be caused by indirect systemic responses.
authors.affiliation: Institut für Tierernährung, Justus-Liebig-Universität Giessen.
authors.forename: K
authors.initials: K
authors.lastname: Krämer
authors.forename: A
authors.initials: A
authors.lastname: Markwitan
authors.forename: J
authors.initials: J
authors.lastname: Pallauf
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism*; D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D001127:Arginine Vasopressin / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002478:Cells, Cultured; D003470:Culture Media; D003907:Dexamethasone / Q000494:pharmacology; D004075:Diglycerides / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D008668:Metallothionein / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000502:physiology*; D010972:Platelet Activating Factor / Q000494:pharmacology; D051381:Rats; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015032:Zinc / Q000494:pharmacology
chemical_list: D003470:Culture Media; D004075:Diglycerides; D010716:Phosphatidylinositols; D010972:Platelet Activating Factor; C044295:1,2-dioctanoylglycerol; D000804:Angiotensin II; D001127:Arginine Vasopressin; D000001:Calcimycin; D003907:Dexamethasone; D008668:Metallothionein; D000469:Alkaline Phosphatase; D015032:Zinc; D013755:Tetradecanoylphorbol Acetate