Protective effect of metallothionein on cadmium toxicity in isolated rat hepatocytes

An isolated rat hepatocyte preparation was used to study the cellular toxicity of cadmium and the protective effects of metallothionein on cadmium-induced toxicity. Exposure of primary suspension cultures of isolated rat hepatocytes to Cd2+ (0-35.7 microM) for 15 min resulted in a dose-dependent reduction in the synthesis of cellular proteins during a subsequent 6 h incubation. Such inhibition could not be correlated with cellular lethality or gross membrane damage. Pre-induction of metallothionein in hepatocytes by zinc treatment in vivo of donor rats protected hepatocytes in vitro from cadmium-induced inhibition of protein synthesis. The protective effects in zinc-pre-induced hepatocytes are not due to alterations in the level of total cellular cadmium, but could be accounted for by the redistribution of intracellular cadmium in the presence of high levels of zinc-metallothionein. The data suggest that metallothionein exerts its protective effect by a kinetic detoxification mechanism, i.e. a decrease in reactive intracellular cadmium.

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Identification of genes early responsive to gamma-irradiation in isolated rat hepatocytes in vitro and rat liver in vivo by cDNA array gene expression analysis

Zeitschrift für Gastroenterologie ◽

10.1055/s-2005-920060 ◽

2005 ◽

Vol 43 (05) ◽

Author(s):

DS Batusic ◽

H Christiansen ◽

B Saile ◽

RM Hermann ◽

J Dudas ◽

...

Keyword(s):

Gene Expression ◽

Gamma Irradiation ◽

Rat Liver ◽

Gene Expression Analysis ◽

Cdna Array ◽

Rat Hepatocytes ◽

Isolated Rat Hepatocytes ◽

Isolated Rat

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Effects of in vivo and in vitro Alcohol Administration on Insulin Binding and Glycogenesis in Isolated Rat Hepatocytes

Annals of Nutrition and Metabolism ◽

10.1159/000176692 ◽

1983 ◽

Vol 27 (4) ◽

pp. 313-319 ◽

Cited By ~ 15

Author(s):

R.M. Rifkin ◽

W.W. Todd ◽

D.R. Toothaker ◽

A. Sussman ◽

M. Trowbridge ◽

...

Keyword(s):

Rat Hepatocytes ◽

Insulin Binding ◽

Isolated Rat Hepatocytes ◽

Alcohol Administration ◽

Isolated Rat

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Assessement of the relation between the initial viability and the attachment of freshly isolated rat hepatocytes used for the in vivo/in vitro DNA repair assay (UDS)

Mutation Research/Environmental Mutagenesis and Related Subjects ◽

10.1016/0165-1161(93)90013-p ◽

1993 ◽

Vol 291 (1) ◽

pp. 21-27 ◽

Cited By ~ 13

Author(s):

Rolf Fautz ◽

Birgitta Husein ◽

Elena Efstathiou ◽

Claudia Hechenberger-Freudl

Keyword(s):

Dna Repair ◽

Rat Hepatocytes ◽

Isolated Rat Hepatocytes ◽

Freshly Isolated Rat Hepatocytes ◽

Isolated Rat

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Metabolism of thyroid hormones in isolated rat hepatocytes: studies on the influences of carbamazepine and phenytoin

Acta Endocrinologica ◽

10.1530/acta.0.1040479 ◽

1983 ◽

Vol 104 (4) ◽

pp. 479-484 ◽

Cited By ~ 2

Author(s):

Sylvi Aanderud ◽

Jarle Aarbakke ◽

Johan Sundsfjord

Keyword(s):

Thyroid Hormones ◽

Cellular Uptake ◽

Incubation Medium ◽

Rat Hepatocytes ◽

Isolated Rat Hepatocytes ◽

Isolated Rat

Abstract. The in vitro handling of thyroid hormones was studied in isolated rat hepatocytes by measuring 1) the cellular uptake of T4, 2) the conversion of T4 to T3 and 3) the degradation of T4 and T3. The in vitro conversion of T4 to T3 increased significantly by adding ethanol 2% or carbamazepine (CBZ) 400 μm in ethanol 2% to the incubation medium. As there was no difference between ethanol and CBZ/ethanol on the T3 formation, this effect was probably caused by ethanol. The T3 formation was unaffected by phenytoin (PHT) in conc. up to 400 μm, while propylthiouracil (PTU) 100 and 400 μm inhibited the conversion completely. The T4 to T3 conversion in hepatocytes from rats pretreated with CBZ or PHT for 2 weeks was not significantly different from untreated controls. The cellular uptake of T4 was reduced by about 30% in the presence of PHT and unaltered by CBZ and ethanol. The degradation of T4 and T3 was not influenced by the in vitro addition of CBZ or PHT, nor was the degradation of T4 and T3 significantly different from untreated controls in hepatocyte suspensions from CBZ or PHT pretreated rats. Our findings suggest that the handling of thyroid hormones in isolated rat hepatocytes is not influenced by the in vitro or in vivo exposure to CBZ or PHT.

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The Hep G2 Cell Line as a Possible Alternative to Isolated Hepatocytes in Cytotoxicity Studies

Alternatives to Laboratory Animals ◽

10.1177/026119298801600105 ◽

1988 ◽

Vol 16 (1) ◽

pp. 16-22

Author(s):

Marina Marinovich ◽

Jose L. Lorenzo ◽

Liliana M. Flaminio ◽

Agnese Granata ◽

Corrado L. Galli

Keyword(s):

Cell Line ◽

Rat Hepatocytes ◽

Prostaglandin E ◽

Human Hepatoma ◽

Hep G2 ◽

Hep G2 Cells ◽

Isolated Rat Hepatocytes ◽

G2 Cells ◽

Isolated Rat

The hepatotoxicity of carbon tetrachloride (CC14) was evaluated in vitro in freshly isolated rat hepatocytes and in the human hepatoma cell line, Hep G2. Toxicity was assessed by the leakage of cytosolic enzymes (lactate dehydrogenase and aspartate aminotransferase) and cell viability (trypan blue exclusion). The established human cells were less sensitive to CCl4-induced injury; higher doses of the toxic agent and longer incubation times were necessary to elicit cell damage. Micromolar concentrations of prostaglandin E2 significantly decreased enzyme leakage in both Hep G2 cells and rat hepatocytes challenged with CC14; a stable derivative of prostacyclin (ZK 36374) was ineffective. These results suggest that human hepatoma Hep G2 cells may represent a valid alternative to isolated rat hepatocytes for an initial approach to the in vitro evaluation of cell toxicity.

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Acellular liver matrix improves the survival and functions of isolated rat hepatocytes cultured in vitro.

International Journal of Molecular Medicine ◽

10.3892/ijmm.14.4.511 ◽

2004 ◽

Cited By ~ 1

Author(s):

Patrizia Burra ◽

Silvia Tomat ◽

Maria Teresa Conconi ◽

Carlo Macchi ◽

Francesco P Russo ◽

...

Keyword(s):

Rat Hepatocytes ◽

Isolated Rat Hepatocytes ◽

Isolated Rat

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Comparative metabolism of the antiviral dimer 3'-azido-3'-deoxythymidine-P-2',3'-dideoxyinosine and the monomers zidovudine and didanosine by rat, monkey, and human hepatocytes.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.41.11.2502 ◽

1997 ◽

Vol 41 (11) ◽

pp. 2502-2510 ◽

Cited By ~ 4

Author(s):

X R Pan-Zhou ◽

E Cretton-Scott ◽

X J Zhou ◽

M Y Xie ◽

R Rahmani ◽

...

Keyword(s):

High Performance ◽

Rat Hepatocytes ◽

Human Hepatocytes ◽

Metabolic Fate ◽

Extracellular Medium ◽

Isolated Rat Hepatocytes ◽

Cell Extracts ◽

Comparative Metabolism ◽

Isolated Rat

AZT-P-ddI is an antiviral heterodimer composed of one molecule of 3'-azido-3'-deoxythymidine (AZT) and one molecule of 2',3'-dideoxyinosine (ddI) linked through their 5' positions by a phosphate bond. The metabolic fate of the dimer was studied with isolated rat, monkey, and human hepatocytes and was compared with that of its component monomers AZT and ddI. Upon incubation of double-labeled [14C]AZT-P-[3H]ddI in freshly isolated rat hepatocytes in suspension at a final concentration of 10 microM, the dimer was taken up intact by cells and then rapidly cleaved to AZT, AZT monophosphate, ddI, and ddI monophosphate. AZT and ddI so formed were then subject to their respective catabolisms. High-performance liquid chromatography analyses of the extracellular medium and cell extracts revealed the presence of unchanged dimer, AZT, 3'-azido-3'-deoxy-5'-beta-D-glucopyranosylthymidine (GAZT), 3'-amino-3'-deoxythymidine (AMT), ddI, and a previously unrecognized derivative of the dideoxyribose moiety of ddI, designated ddI-M. Trace extracellular but substantial intracellular levels of the glucuronide derivative of AMT (3'-amino-3'-deoxy-5'-beta-D-glucopyranosylthymidine [GAMT]) were also detected. Moreover, the extent of the formation of AMT, GAZT, and ddI-M from the dimer was markedly lower than that with AZT and ddI alone by the hepatocytes. With hepatocytes in primary culture obtained from rat, monkey, and human, large interspecies variations in the metabolism of AZT-P-ddI were observed. While GAZT and ddI-M, metabolites of AZT and ddI, respectively, as well as AZT 5'-monophosphate (MP) and ddI-MP were detected in the extracellular media of all species, AMT and GAMT were produced only by rat and monkey hepatocytes. No such metabolites were formed by human hepatocytes. The metabolic fate of the dimer by human hepatocytes was consistent with in vivo data recently obtained from human immunodeficiency virus-infected patients.

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