Effects of the peroxisome proliferator di(2-ethylhexyl)phthalate on cell turnover and peroxisome proliferation in primary chick embryo hepatocytes

The peroxisome proliferator (PP) di(2-ethylhexyl)phthalate (DEHP) is widely used as a plasticizer and can contaminate air, water, and soil. As yet, no data have been published on its potential to induce changes in cell growth of nonmammalian hepatocytes. In the present study, the effects of DEHP on cell turnover and induction of peroxisome proliferation were evaluated in primary hepatocyte cultures from chick embryos. Cells were treated after attachment with 0, 25, 50, 75, and 100 µM DEHP for up to 96 h. S-phase increased significantly (p < 0.01) from a background level of 5.5 ± 0.1% in solvent-control hepatocytes to a maximum level of 7.1 ± 0.1% in cells exposed for 48 h to 100 µM DEHP and decreased to near 6% by 96 h. Lower (p < 0.05) levels of induction were seen at 50 and 75 µM DEHP. Spontaneous apoptosis showed a slight (p < 0.05) decrease in hepatocytes treated with ≥75 µM dosages, as measured at 72 to 96 h. Induction of peroxisome proliferation was observed for cultures treated with ≥75 µM dosages at 48 h onwards. The results of the present study indicate that avian species may be responsive to the effects of PPs and may thus be affected by the presence of DEHP in the environment, but that this species is less sensitive than rodents.