Epigallocatechin-3-gallate induces cytokine production in mast cells by stimulating an extracellular superoxide-mediated calcium influx.

The green tea polyphenol (-)-epigallocatechin-3-O-gallate (EGCG) has various biological activities, including anti-inflammatory, anti-neoplastic, anti- and pro-apoptotic, and neuroprotective effects. Although these are often associated with increased intracellular reactive oxygen species (ROS) and Ca(2+) levels, their involvement in biological effects is poorly understood. Here we report that EGCG induces cytokine production in mast cells via Ca(2+) influx and ROS generation. EGCG at concentrations of ≥50 μM induced interleukin-13 and tumor necrosis factor-α production in RBL-2H3 and bone marrow-derived mast cells. The effects were dependent on extracellular Ca(2+), and EGCG induced Ca(2+) release from intracellular stores and Ca(2+) influx. Ca(2+) influx was suppressed by 2-aminoethoxydiphenyl borate, an inhibitor of store-operated Ca(2+) (SOC) channels, including Ca(2+) release-activated Ca(2+) channels and transient receptor potential canonical channels. EGCG failed to induce Ca(2+) influx through SOC channels. EGCG-activated Ca(2+) channels were genetically and pharmacologically distinct from Ca(v)1.2 L-type Ca(2+) channels, another route of Ca(2+) influx into mast cells. EGCG evoked release of superoxide (O(2)(·-)) into the extracellular space. Exogenous superoxide dismutase, but not catalase, inhibited EGCG-evoked Ca(2+) influx and cytokine production, indicating that extracellular O(2)(·-) regulates these events. EGCG can serve as a powerful tool for studying O(2)(·-)-regulated Ca(2+) channels, which may be selectively involved in the regulation of cytokine production but have yet to be elucidated. Copyright Â