Cs. Huang et al., Vanadium-induced nuclear factor of activated T cells activation through hydrogen peroxide, J BIOL CHEM, 276(25), 2001, pp. 22397-22403
The present study investigated the role of reactive oxygen species (ROS) in
activation of nuclear factor of activated T cells (NFAT), a pivotal transc
ription factor responsible for regulation of cytokines, by vanadium in mous
e embryo fibroblast PW cells or mouse epidermal Cl 41 cells. Exposure of ce
lls to vanadium led to the transactivation of NFAT in a time- and dose-depe
ndent manner. Scavenging of vanadium-induced H2O2 with Nacety-L-cyteine (a
general antioxidant) or catalase (a specific H2O2 inhibitor) or the chelati
on of vanadate with deferoxamine, resulted in inhibition of NFAT activation
. In contrast, an increase in H2O2 generation by the addition of superoxide
dismutase or NADPH enhanced vanadium-induced NEAT activation. This vanadat
e-mediated H2O2 generation was verified by both electron spin resonance and
fluorescence staining assay. These results demonstrate that H2O2 plays an
important role in vanadium-induced NFAT transactivation in two different ce
ll types. Furthermore, pretreatment of cells with nifedipine, a calcium cha
nnel blocker, inhibited vanadium-induced NFAT activation, whereas A23187 an
d ionomycin, two calcium ionophores, had synergistic effects with vanadium
for NFAT induction, Incubation of cells with cyclosporin A (CsA), a pharmac
ological inhibitor of the phosphatase calcineurin, blocked vanadium-induced
NFAT activation. Ah data show that vanadium induces NFAT activation not on
ly through a calcium-dependent and CsA-sensitive pathway but also involved
H2O2 generation, suggesting that H2O2 may be involved in activation of calc
ium-calcineurin pathways for NFAT activation caused by vanadium exposure.