Inhibitory effect of anethole on T-lymphocyte proliferation and interleukin-2 production through down-regulation of the NF-AT and AP-1

Abstract

Anethole is a naturally occurring alkenylbenzene found in a variety of foods and essential oils. In the present study, we investigated the effect of anethole on T-cell function and the regulatory mechanism of its effect. Direct addition of anethole to B6C3F1 mouse splenocyte cultures produced a concentration-dependent inhibition of the lymphoproliferative response to concanavalin A stimulation. Anethole inhibited phorbol 12-myristate 13-acetate (PMA) plus ionomycin (Io)-induced interleukin-2 (IL-2) mRNA expression and protein secretion in EL4 mouse T-cells as determined by quantitative/competitive RT-PCR and ELISA, respectively. To further characterize the mechanism for the transcriptional regulation of IL-2, an electrophoretic mobility shift assay was performed to evaluate the binding activity of the nuclear factor of activated T-cells (NF-AT), activator protein-1 (AP-1), nuclear factor-kappaB (NF-kappaB), and octamer binding protein (Oct) in PMA/Io-stimulated EL4 cells. Anethole decreased the NF-AT and AP-1 binding activity, but no significant effect was observed on NF-kappaB or Oct binding activity. These results suggest that anethole suppress T-cell proliferation and IL-2 production and that the inhibition is mediated, at least in part, through the down-regulation of NF-AT and AP-1.