Lead (Pb) is an environmental toxicant that can induce structural and functional abnormalities of multiple organ systems, including the central nervous and the immune systems. The aim of this study was to evaluate the effects of extracellular Pb supplementation on the cellular content of the metal and on the proliferation and the survival of normal rat fibroblasts. We found that the concentration of Pb in the culture medium was 0.060 microM and the normal Pb concentration in rat fibroblasts was 3.1 +/- 0.1 ng/10(7) cells. Then we exposed the cells to increasing concentration of Pb (as Pb acetate) from 0.078-320 microM. We observed a dose-dependent inhibition of cell proliferation after 48 h, which was already apparent at a concentration of 0.312 microM (p = 0.122) and became statistically significant for concentration higher than 0.625 microM (p = 0.0003 at 5 microM). Cell proliferation was completely compromised at 320 microM Pb total inhibition of cell proliferation. To investigate the mechanisms of Pb-mediated inhibition of cell proliferation, we evaluated the occurrence of apoptosis in the same cells and found that cytosolic DNA fragments, hallmark of apoptotic cell death, increased significantly at Pb concentrations from 2.5-10.0 microM. The occurrence of apoptosis was also confirmed by FACS analysis which showed the appearance of a subdiploid peak at Pb concentrations from 5-20 microM. The distribution of cells in the cell cycle showed a dose-dependent accumulation of cells in the G0/G1 phase mainly compensated by a decrease in the percentage of cells in the S phase. In conclusion, our results demonstrate that induction of apoptosis contributes to the Pb-induced inhibition of cell proliferation in rat fibroblasts.