Z Gastroenterol 2005; 43 - 100
DOI: 10.1055/s-2005-869747

In vitro model system for study of heptotoxicity

A Petri 1, M Radács 5, L Gáspár 2, A Juhász 3, Z Valkusz 2, E László Kókai 4, M Gálfi 5
  • 1Department of Surgery, Faculty of Medicine, Szeged University
  • 2Department of Endocrinology, Faculty of Medicine, Szeged University
  • 3Department of Psychiatry, Faculty of Medicine, Szeged University
  • 4Department of Dentistry, Faculty of Medicine, Szeged University
  • 5Environmental Sciences Team, Deptartment of Biology, Juhász Gyula Teacher Training Faculty Szeged University

The hepatocells, as they perform the maintenance of homeostasis, are not always in direct contact with one another, but carry out a continuous communication. The chemical language of communication is maintained via various biological products, changes of cellcycle processes. Different agents, as halogenated hydrocarbones, induce the different biological processes on the liver, which can be the markers of these cells.

Our aims were to investigate the effects of subtoxical doses chlorobenzenes on in vitro hepatocyte modells, obtained in various doses, and to compare their efficacy on the cellproliferation and cellfunction.

We used (150g male) Wistar rats and human liver samples. Following an enzymatic (Trypsin, Collagenase, DNase I and II, Dispase) and mechanical dissociation of the tissues we generated monolayer hepatocyte (Hep) cultures. The cultures were kept in a CO2 thermostat at 37C, in cell-specific media, until the appearance of confluent monolayers.

The untreated cultures were used as the absolute control (K) group. The positive controls were in the presence of benz(c)acridines (1mg/ml), and the negative controls were treated by the 1.5% ethanol. The experimental samples were treated chlorobenzene: (0.01–10µg/ml). During the experiments were used the antiproliferative agents: crocine/100µg/ml/(/Sigma-Aldrich/20µM). After the treatment were detected the markers of cellproliferation, as a protein and DNA content of the different cell cultures, and the number of tumorous clones. The physiological parameters were controlled by the enzymactivity of hepatocells (LDH, ASAT, ALAT).

From our results showed that, the extremly low doses of chlorobenzenes can not induced changes of the measured parameters. The cellproliferating-cascade on the hepatocytes was observed after the treating with higher ClB doses (>1µg/ml). The celltransformation was effective in ClB induced processes. The benz(c)acridines caused cellproliferation on hepatocytes in all case. The antiproliferative agents (crocine and retinoic acide) significantly reduced the celltransformation in all models. The modulation of enzymactivity was correlated by the cellproliferating effects.

This modelsystem is suitable for the study the hepatotoxic and cellproliferation processes caused disease by the liver.

This research was supported by: ETT 270/2003; ETT 61/2003