Biphasic effect of iron on human intestinal Caco-2 cells: Early effect on tight junction permeability with delayed onset of oxidative cytotoxic damage

Treatment of differentiated human intestinal Caco-2 cells with Fe(H) ascorbate altered fight junction permeability in a dose and time-dependent way for up to 3 hr of treatment. Upon iron removal and transfer to complete culture medium, the effect was reversible up to 10 M Fe(II), while at higher concentrations a late phase toxic effect was observed. Reduction of intracellular energy abolished the short term effect of iron on fight junction permeability without affecting its cellular uptake, suggesting that active processes, other than transport, were involved. The short term effect of iron on the permeability of fight junctions did not appear to result from the generation of reactive oxygen species, as it was not prevented by antioxidant treatment under normal energy conditions. Conversely, the late phase effect, leading to both apoptosis and necrosis during the 24 hr following iron removal, could be reduced by antioxidant treatment and was exacerbated by GSH depletion. Iron induced oxidative stress may therefore be responsible for membrane damage and cellular death occurring in the late phase. The reported effects of iron on intestinal tight junction permeability followed by more widespread cytotoxicity from oxidative events should be considered in light of the extensive use of iron supplementation in different phases of human life.