Toxicological effects of iron on intestinal cells.
The aim of the present study was to investigate whether iron, which is involved in the formation of free radicals in vitro, can initiate cellular injury in human intestinal cells. The effects of various concentrations of iron were studied in preconfluent, colonic-cancerogenous cells, and also in postconfluent, differentiating cells. Cellular damage was assessed using cell proliferation (serial cell counting), tetrazolium dye (MTT) uptake, lactate dehydrogenase (LDH) release and apoptosis studies based on caspase-3 activities. Also the activities of the major antioxidative enzymes, superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) were measured after the cells had been exposed to iron. Our results indicated that preconfluent cells were more susceptible to iron toxicity, as assessed by a significant reduction in cell proliferation and MTT uptake in a concentration-dependent manner compared to the control. However, no evidence for MTT uptake was observed in postconfluent cells. Caspase-3 activity, an indicator of cell apoptosis, considerably increased in preconfluent cells at high iron levels compared to the control (p < 0.05), whereas postconfluent cells were not significantly affected. LDH release was similar for both groups and was significantly higher than the control at 900 microM iron and above. SOD activities were not affected by iron in either group, whereas GPx was considerably higher in iron-treated cells in both groups compared with the control (because of relatively high standard deviations this effect was not significant). In conclusion we suggest that iron exerts its toxic effects intracellularly especially in preconfluent Caco-2 cells, whereas only high iron doses were able to alter the viability of differentiating, enterocyte-like cells.
Zödl
Bettina
B
Department of Physiology, University of Vienna, Faculty of Medicine, Vienna, Austria.
Sargazi
Mansour
M
Zeiner
Michaela
M
Roberts
Norman B
NB
Steffan
Ilse
I
Marktl
Wolfgang
W
Ekmekcioglu
Cem
C
D017209:Apoptosis; D001681:Biological Assay; D018938:Caco-2 Cells; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000166:cytology / Q000187:drug effects*; D007501:Iron / Q000378:metabolism / Q000633:toxicity*; D010088:Oxidoreductases / Q000378:metabolism
D007501:Iron; D010088:Oxidoreductases; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases