In vitro lead-induced cell toxicity and cytoprotective activity of fetal calf serum in human fibroblasts.
The underlying mechanisms by which lead ions produce their deleterious effects prior to the onset of clinical symptoms are incompletely understood. This study aimed to assess lead-induced cell toxicity mechanisms by focusing on the effects of the metal on cell growth, DNA synthesis, cellular ATP, intracellular hexosaminidase activity and lysosomal function, and examine the possible cytoprotective role of fetal calf serum (FCS). Several human dermal cultured fibroblast lines were exposed to Pb (400 microM) for 1-6 days with 2, 5, and 10% FCS. The earliest toxic effect of Pb was significant inhibition of DNA synthesis after 24 h direct exposure; this harmful effect was not progressive during the first 3 days, but worsened clearly on the 4th day regardless of the FCS concentration. Atime-dependent depletion of intracellularATP content was also caused by ionic lead, thereby compromising the cell energy charge which precedes cell death. Fibroblast growth was progressively and significantly inhibited from day 2 onwards; the greatest noxious effect was observed in the presence of 2% FCS: 49% reduction in cell proliferation after 5 days. Lead salts produced loss of cell adhesion to the culture dish which worsened from the 2nd day and was more pronounced when FCS in growth medium was decreased. Toxic actions on lysosomal membrane integrity provoked a decrease in neutral red uptake (NRU) which was exposure time-dependent and more marked with 2% FCS. In contrast, increased relative NRU (to 20% at 4 days), suggestive of endocytosis-induced lysosome enlargement, was observed in Pb-exposed cells. Intracellular hexosaminidase activity was not negatively affected until 5 days after exposure to Pb salts. FCS had a significant cytoprotective effect on Pb-induced toxicity.
Dominguéz
Carmen
C
Biochemistry and Molecular Biology Center, Vall d 'Hebron Hospitals, Barcelona, Spain. cdomin@hg.vhebron.es
Solé
Eduard
E
Fortuny
Anna
A
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001769:Blood / Q000378:metabolism*; D002417:Cattle; D002448:Cell Adhesion; D016923:Cell Death; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007854:Lead / Q000494:pharmacology; D009174:Mycoplasma / Q000378:metabolism; D016133:Polymerase Chain Reaction; D013997:Time Factors; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
D003470:Culture Media; D007854:Lead; D000255:Adenosine Triphosphate; D001619:beta-N-Acetylhexosaminidases